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What Is Yeast?

Yeast are a group of unicellular fungi a few species of which are commonly used to leaven bread and ferment alcoholic beverages. Most yeasts belong to the division Ascomycota. A few yeasts, such as Candida albicans can cause infection in humans. More than one-thousand species of yeasts have been described. The most commonly used yeast is Saccharomyces cerevisiae, which was domesticated for wine, bread and beer production thousands of years ago. See Yeast (baking).

Yeast physiology can be either obligately aerobic or facultatively fermentative. There is no known obligately anaerobic yeast. In the absence of oxygen, fermentative yeasts produce their energy by converting sugars into carbon dioxide and ethanol (alcohol). In brewing, the ethanol is used, while in baking the carbon dioxide raises the bread and the ethanol evaporates.

An example with glucose as the substrate is

C6H12O6 (glucose) →2C2H5OH + 2CO2 Yeasts can reproduce asexually through budding or sexually through the formation of ascospores. During asexual reproduction a new bud grows out of the parent yeast when the condition is right, then after the bud reaches an adult size, it separates from the parent yeast. Under low nutrient conditions, yeasts that are capapable of sexual reproduction will form ascospores. Yeasts that are not capable of going through the full sexual cycle are classified in the genus Candida.

Yeasts for leavening bread may be produced commercially or caught from the environment. Many yeasts can be isolated from sugar-rich environmental samples. Some good examples include fruits and berries (such as grapes, apples or peaches), exudates from plants (such as plant saps or cacti). Some yeasts are found in association with insects.

The use of potatoes, water from potato boiling, eggs, or sugar in a bread dough accelerates the growth of yeasts. Salt and fats such as butter slow yeast growth down. A common medium used for the cultivation of yeasts is called potato dextrose agar (PDA) or potato dextrose broth. Potato extract is made by autoclaving cut-up potatoes with water for 5 to 10 minutes and then decanting off the broth. Dextrose (glucose) is then added (10 g/L), and the medium is sterilized by autoclaving.

Yeast fermentations comprise the oldest and largest application of microbial technology. They are used for beer and wine fermentations and bread production. Beer brewers classify yeasts as top-fermenting and bottom-fermenting. This distinction was introduced by the Dane Emil Christian Hansen.

Top-fermenting yeasts (so-called because they float to the top of the beer) can produce higher alcohol concentrations and prefer higher temperatures. An example is Saccharomyces cerevisiae, known to brewers as ale yeast. They produce fruitier, sweeter, real ale type beers. Bottom-fermenting yeasts ferment more sugars leaving a crisper taste and work well at low temperatures. An example is Saccharomyces uvarum, formerly known as Saccharomyces carlsbergensis. They are used in producing lager-type beers. Brewers of wheat beers often use varieties of Torulaspora delbrueckii.

Winemakers use a variety of different yeasts depending on the type of wine and the condition of the grapes. Too high a sugar or alcohol concentration slows the growth of yeast, so for very ripe grapes with lots of sugar he or she would use a yeast tolerant of those conditions. If the yeast dies before all the fermentable sugar has been converted to alcohol, the result is a "stuck" fermentation. Some yeast is chosen because it tends to develop certain aromas, such as the distinctive "banana" smells of Beaujolais from Georges Duboeuf. Wild yeast are naturally present on the skins of grapes, so grape juice will spontaneously ferment unless the wild yeast are arrested by cold temperature or sulfates. Depending on the strain of indigenous yeast, the result may be unpalatable or possibly more complex than if a single cultured strain were used. In general, natural yeasts are riskier than cultured, and tend to be used by tradition-oriented, Old World-style winemakers.

Saccharomyces cerevisiae is also known as budding or baker's yeast. It is used as a model organism by biologists studying genetics and molecular biology (in particular the cell cycle) because it is easy to culture but as a eukaryote, it shares the complex internal cell structure of plants and animals.

Saccharomyces cerevisiae was the first eukaryotic genome that was completely sequenced. The yeast genome database [1] (http://www.yeastgenome.org/) is highly annotated and remains a very important tool for developing basic knowledge about the function and organization of eukaryotic cell genetics and physiology. Another important S. cerevisiae database is maintained by the Munich Information Center for Protein Sequences [2] (http://mips.gsf.de/genre/proj/yeast/index.jsp).

Another important experimental model is Schizosaccharomyces pombe or fission yeast.

Candida albicans, a diploid sexual fungus (a form of yeast) is the causal agent of opportunistic infections in humans, the most common being oral and vaginal infections. Systemic fungal infections have emerged as important causes of morbidity and mortality in immunocompromised patients (e.g., AIDS, cancer chemotherapy, organ or bone marrow transplantation). In addition, hospital-related infections in patients not previously considered at risk (e.g. patients on an intensive care unit) have become a cause of major health concern.

Among the many organisms that live in the human mouth and digestive tract is the yeast Candida albicans, which under normal circumstances lives in 80 percent of the human population with no harmful effects. However, overgrowth results in candidiasis. Candidiasis is often observed in immunocompromised individuals such as HIV-positive patients. Candidiasis also may occur in the blood and in the genital tract. Candidiasis is commonly known as "thrush", and is a common condition that is usually easily cured in people who are not immunocompromised. To infect host tissue, the usual unicellular yeast-like form of Candida albicans reacts to environmental cues and switches into an invasive, multicellular filamentous form. This switching between two cell-types is known as dimorphism.

FEBS Lett, 2000 Apr 7, 471(1), 103 - 7
Interaction between the critical aromatic amino acid residues Tyr(352) and Phe(504) in the yeast Gal2 transporter; Kasahara T et al.; Three critical aromatic sites have been identified in the yeast galactose transporter Gal2: Tyr(352) at the extracellular boundary of putative transmembrane segment (TM) 7, Tyr(446) in the middle of TM10 and Phe(504) in the middle of TM12 . The relationship between these sites was investigated by random mutagenesis of each combination of two of the three residues . Galactose transport-positive clones selected by plate assays encoded Tyr(446) and specific combinations of aromatic residues at sites 352 and 504 . Double-site mutants containing aromatic residues at these latter two positions showed either essentially full galactose transport activity (Phe(352)Trp(504) and Trp(352)Trp(504)) or no significant activity (Phe(352)Tyr(504) and Trp(352)Tyr(504)), whereas single-site mutants showed markedly reduced activity . These results are indicative of a specific interaction between sites 352 and 504 of Gal2.

Mol Microbiol, 2000 Apr, 36(1), 163 - 73
The NC2 repressor is dispensable in yeast mutated for the Sin4p component of the holoenzyme and plays roles similar to Mot1p in vivo; Lemaire M et al.; NC2 (Dr1/DRAP1) and Mot1p are global repressors of transcription that have been isolated in both Saccharomyces cerevisiae and humans . NC2 is a dimeric histone-fold complex that represses RNA polymerase II transcription through binding to TBP and inhibition of TFIIA and TFIIB . Mot1p is an ATPase that removes DNA-bound TBP upon ATP hydrolysis . In this work, we studied the core promoter specificity of NC2 in vivo using a strain that carries mutated NC2beta activity . We show that NC2, like Mot1p, is required for transcription of the HIS3 and HIS4 TATA-less core promoters . Furthermore, whereas neither Mot1p nor NC2 appear to function as repressors of the HIS3 gene in cells growing exponentially in glucose, we find that both are required for repression of the HIS3 TATA promoter when cells go through the diauxic shift . Thus, the activity of these factors is similarly regulated depending upon the physiological conditions, and it appears that core promoters activated or repressed by them in vivo might be distinguishable by whether or not they contain a canonical TATA sequence . Finally, although NC2 is an essential factor for yeast viability, we isolated a mutation in a non-essential component of the holoenzyme, Sin4p, that bypasses the requirement for NC2.

Genes Cells, 2000 Mar, 5(3), 221 - 33
A human homologue of yeast anti-silencing factor has histone chaperone activity; Munakata T et al.; BACKGROUND: Structural changes in chromatin play essential roles in regulating eukaryotic gene expression . Silencing, potent repression of transcription in Saccharomyces cerevisiae, occurs near telomeres and at the silent mating-type loci, as well as at rDNA loci . This type of repression relates to the condensation of chromatin that occurs in the heterochromatin of multicellular organisms . Anti-silencing is a reaction by which silenced loci are de-repressed . Genetic studies revealed that several factors participate in the anti-silencing reaction . However, actions of factors and molecular mechanisms underlying anti-silencing remain unknown . RESULTS: Here we report the functional activity of a highly evolutionarily conserved human factor termed CIA (CCG1-interacting factor A), whose budding yeast homologue ASF1 has anti-silencing activity . Using yeast two-hybrid screening, we isolated histone H3 as an interacting factor of CIA . We also showed that CIA binds to histones H3/H4 in vitro, and that the interacting region of histone H3 is located in the C-terminal helices . Considering the functional role of CIA as a histone-interacting protein, we found that CIA forms a nucleosome-like structure with DNA and histones . CONCLUSIONS: These results show that human CIA, whose yeast homologue ASF1 is an anti-silencing factor, possesses histone chaperone activity . This leads to a better understanding of the relationship between chromatin structural changes and anti-silencing processes.

Genes Cells, 2000 Mar, 5(3), 169 - 90
Large-scale screening of intracellular protein localization in living fission yeast cells by the use of a GFP-fusion genomic DNA library; Ding DQ et al.; BACKGROUND: Intracellular localization is an important part of the characterization of a gene product . In an attempt to search for genes based on the intracellular localization of their products, we constructed a green fluorescent protein (GFP)-fusion genomic DNA library of S . pombe . RESULTS: We constructed the S . pombe GFP-fusion genomic DNA library by fusing, in all three reading frames, random fragments of genomic DNA to the 5' end of the GFP gene in such a way that expression of potential GFP-fusion proteins would be under the control of the own promoters contained in the genomic DNA fragments . Fission yeast cells were transformed with this plasmid library, and microscopic screening of 49 845 transformants yielded 6954 transformants which exhibited GFP fluorescence, of which 728 transformants showed fluorescence localized to distinct intracellular structures such as the nucleus, the nuclear membrane, and cytoskeletal structures . Plasmids were isolated from 516 of these transformants, and a determination of their DNA sequences identified 250 independent genes . The intracellular localizations of the 250 GFP-fusion constructs was categorized as an image database; using this database, DNA sequences can be searched for based on the localizations of their products . CONCLUSIONS: A number of new intracellular structural components were found in this library . The library of GFP-fusion constructs also provides useful fluorescent markers for various intracellular structures and cellular activities, which can be readily used for microscopic observation in living cells.

Int J Syst Evol Microbiol, 2000 Mar, 50 Pt 2, 931 - 6
Non-radioactive dot-blot DNA reassociation for unequivocal yeast identification; Cardinali G et al.; Unequivocal and rapid classification of yeast cultures cannot be accomplished exclusively on the basis of unstable phenotypic traits, but requires molecular tests relating to the whole genome (or the largest possible portion of it) . DNA-DNA reassociation meets this requirement, although many procedures proposed for calculating overall sequence similarity are expensive and time-consuming, thus restricting the possibility of unequivocal classification to a few specialized laboratories . A novel method, based on non-radioactive dot-blot hybridization of whole genomic DNA, has shown high and reproducible proportionality between the detected signal and the amount of double-stranded DNA effectively present on the membrane . This procedure has been optimized to obtain, within two working days, DNA relatedness values between unknown cultures and the type strains of the species previously indicated by a few conventional tests . The effective ability of the method to discriminate strains belonging to different species has been tested within taxonomic models consisting of yeast type cultures already certified by spectrophotometric reassociation.

Plant J, 2000 Feb, 21(4), 341 - 9
Modes of interaction between the Arabidopsis Rab protein, Ara4, and its putative regulator molecules revealed by a yeast expression system; Ueda T et al.; Ara4, a member of the Rab/Ypt GTPase family derived from Arabidopsis thaliana, causes severe growth inhibition when expressed in several yeast ypt mutants . Mutational analysis of ARA4 indicated that the Ara4 protein titrates at least three factors in yeast, including the GDP dissociation inhibitor (GDI) . The coexpression of AtGDI1 (Arabidopsis GDI) suppressed the growth defect caused by Ara4 in yeast ypt1, suggesting that Ara4 and AtGDI1 interact in yeast to compensate for the titration of yeast GDI . We screened an Arabidopsis cDNA library for other suppressors that may also interact with Ara4 physiologically . A novel suppressor, SAY1, encoded a hydrophilic protein with two putative coiled-coil regions, which showed partial similarity to the yeast Vps27 protein . To understand the structural requirements of Ara4 for interacting with these molecules, we examined whether AtGDI1 and SAY1 could suppress the growth defect of ypt1 caused by various mutant versions of ARA4 . The results indicated that the interaction between Ara4 and AtGDI1 depends on the conserved C-terminal Cys-motif and Thr44 in the effector domain of Ara4 . In contrast, neither of these motifs is necessary for the interaction between Say1 and Ara4 . This approach provides a powerful method to dissect complex interactions between a GTPase and its regulators.

J Immunol Methods, 2000 Apr 21, 238(1-2), 161 - 72
In vivo selection of single-chain antibodies using a yeast two-hybrid system; Portner-Taliana A et al.; The current methodology for screening libraries of single-chain fragments of immunoglobulin variable domains (sFvs) utilizes bacterial phage systems . We have developed a unique in vivo selection protocol combining a modified yeast two-hybrid assay with a novel prey vector expressing sFvs . The viability of the system is demonstrated with the screen of a sFv library cloned into a yeast two-hybrid prey vector for molecules that target the bait ATF-2, a member of the CREB/ATF family of transcriptional regulatory proteins . The isolated sFv was capable of recognizing ATF-2 in vitro on Western blots and in vivo in mammalian cells.

J Immunol Methods, 2000 Apr 21, 238(1-2), 29 - 43
Phagocytosis of yeast: a method for concurrent quantification of binding and internalization using differential interference contrast microscopy; Bos H et al.; In studies of phagocytosis there is a need to distinguish targets that are internalized by the cell from those that are bound to the cell surface . The present work describes a simple method by which internalized and surface-bound yeast particles can be identified by differential interference contrast microscopy, using trypan blue to stain surface-bound yeast particles . The method has the advantage that both internalized and surface-bound particles can be visualized without the need to switch the illumination source and/or filter sets, thus facilitating concurrent quantitation of binding and internalization . The method was evaluated with the phagocytosis-modulating agents horseradish peroxidase (HRP) and cytochalasin D, using adherent resident macrophages as phagocytic cells . When macrophages are challenged with a particular type of target, they usually bind many more targets than they ingest . It was shown that yeast particles were arrested in the initial binding phase of phagocytosis depending on the region of macrophage plasma membrane where binding sites were formed . Failure of surface-bound yeast particles to trigger internalization was not due to modifications of the yeast particle surface . Nor was it due to binding to non-phagocytic receptors, or low-affinity receptor-ligand interactions . The glycoprotein HRP inhibited only the binding stage of phagocytosis, whereas cytochalasin D, a drug that affects actin polymerization, inhibited both binding and internalization . However, when the yeast particles were pre-incubated in fresh mouse serum, cytochalasin D inhibited only the internalization step . The assay described here may be useful in studies concerned with the function and expression of phagocytosis-mediating surface lectins.

Proc Natl Acad Sci U S A, 2000 Apr 25, 97(9), 4724 - 9
RNA polymerase I transcription factor Rrn3 is functionally conserved between yeast and human; Moorefield B et al.; We have cloned a human cDNA that is related to the RNA polymerase I transcription factor Rrn3 of Saccharomyces cerevisiae . The recombinant human protein displays both sequence similarity and immunological crossreactivity to yeast Rrn3 and is capable of rescuing a yeast strain carrying a disruption of the RRN3 gene in vivo . Point mutation of an amino acid that is conserved between the yeast and human proteins compromises the function of each factor, confirming that the observed sequence similarity is functionally significant . Rrn3 is the first RNA polymerase I-specific transcription factor shown to be functionally conserved between yeast and mammals, suggesting that at least one mechanism that regulates ribosomal RNA synthesis is conserved among eukaryotes.

Mol Cell Biol, 2000 May, 20(9), 3245 - 55
A novel family of cell wall-related proteins regulated differently during the yeast life cycle; Rodriguez-Pena JM et al.; The Saccharomyces cerevisiae Ygr189c, Yel040w, and Ylr213c gene products show significant homologies among themselves and with various bacterial beta-glucanases and eukaryotic endotransglycosidases . Deletion of the corresponding genes, either individually or in combination, did not produce a lethal phenotype . However, the removal of YGR189c and YEL040w, but not YLR213c, caused additive sensitivity to compounds that interfere with cell wall construction, such as Congo red and Calcofluor White, and overexpression of YEL040w led to resistance to these compounds . These genes were renamed CRH1 and CRH2, respectively, for Congo red hypersensitive . By site-directed mutagenesis we found that the putative glycosidase domain of CRH1 was critical for its function in complementing hypersensitivity to the inhibitors . The involvement of CRH1 and CRH2 in the development of cell wall architecture was clearly shown, since the alkali-soluble glucan fraction in the crh1Delta crh2Delta strain was almost twice the level in the wild-type . Interestingly, the three genes were subject to different patterns of transcriptional regulation . CRH1 and YLR213c (renamed CRR1, for CRH related) were found to be cell cycle regulated and also expressed under sporulation conditions, whereas CRH2 expression did not vary during the mitotic cycle . Crh1 and Crh2 are localized at the cell surface, particularly in chitin-rich areas . Consistent with the observed expression patterns, Crh1-green fluorescent protein was found at the incipient bud site, around the septum area in later stages of budding, and in ascospore envelopes . Crh2 was found to localize mainly at the bud neck throughout the whole budding cycle, in mating projections and zygotes, but not in ascospores . These data suggest that the members of this family of putative glycosidases might exert a common role in cell wall organization at different stages of the yeast life cycle.

Mol Cell Biol, 2000 May, 20(9), 3234 - 44
Cid1, a fission yeast protein required for S-M checkpoint control when DNA polymerase delta or epsilon is inactivated; Wang SW et al.; The S-M checkpoint is an intracellular signaling pathway that ensures that mitosis is not initiated in cells undergoing DNA replication . We identified cid1, a novel fission yeast gene, through its ability when overexpressed to confer specific resistance to a combination of hydroxyurea, which inhibits DNA replication, and caffeine, which overrides the S-M checkpoint . Cid1 overexpression also partially suppressed the hydroxyurea sensitivity characteristic of DNA polymerase delta mutants and mutants defective in the "checkpoint Rad" pathway . Cid1 is a member of a family of putative nucleotidyltransferases including budding yeast Trf4 and Trf5, and mutation of amino acid residues predicted to be essential for this activity resulted in loss of Cid1 function in vivo . Two additional Cid1-like proteins play similar but nonredundant checkpoint-signaling roles in fission yeast . Cells lacking Cid1 were found to be viable but specifically sensitive to the combination of hydroxyurea and caffeine and to be S-M checkpoint defective in the absence of Cds1 . Genetic data suggest that Cid1 acts in association with Crb2/Rhp9 and through the checkpoint-signaling kinase Chk1 to inhibit unscheduled mitosis specifically when DNA polymerase delta or epsilon is inhibited.

Mol Cell Biol, 2000 May, 20(9), 3125 - 36
Biochemical and genetic analysis of the mitochondrial response of yeast to BAX and BCL-X(L); Gross A et al.; The BCL-2 family includes both proapoptotic (e.g., BAX and BAK) and antiapoptotic (e.g., BCL-2 and BCL-X(L)) molecules . The cell death-regulating activity of BCL-2 members appears to depend on their ability to modulate mitochondrial function, which may include regulation of the mitochondrial permeability transition pore (PTP) . We examined the function of BAX and BCL-X(L) using genetic and biochemical approaches in budding yeast because studies with yeast suggest that BCL-2 family members act upon highly conserved mitochondrial components . In this study we found that in wild-type yeast, BAX induced hyperpolarization of mitochondria, production of reactive oxygen species, growth arrest, and cell death; however, cytochrome c was not released detectably despite the induction of mitochondrial dysfunction . Coexpression of BCL-X(L) prevented all BAX-mediated responses . We also assessed the function of BCL-X(L) and BAX in the same strain of Saccharomyces cerevisiae with deletions of selected mitochondrial proteins that have been implicated in the function of BCL-2 family members . BAX-induced growth arrest was independent of the tested mitochondrial components, including voltage-dependent anion channel (VDAC), the catalytic beta subunit or the delta subunit of the F(0)F(1)-ATP synthase, mitochondrial cyclophilin, cytochrome c, and proteins encoded by the mitochondrial genome as revealed by {rho(0)} cells . In contrast, actual cell killing was dependent upon select mitochondrial components including the beta subunit of ATP synthase and mitochondrial genome-encoded proteins but not VDAC . The BCL-X(L) protection from either BAX-induced growth arrest or cell killing proved to be independent of mitochondrial components . Thus, BAX induces two cellular processes in yeast which can each be abrogated by BCL-X(L): cell arrest, which does not require aspects of mitochondrial biochemistry, and cell killing, which does.

Mol Cell Biol, 2000 May, 20(9), 3027 - 36
A role for the Hsp40 Ydj1 in repression of basal steroid receptor activity in yeast; Johnson JL et al.; In addition to its roles in translocation of preproteins across membranes, Ydj1 facilitates the maturation of Hsp90 substrates, including mammalian steroid receptors, which activate transcription in yeast in a hormone-dependent manner . To better understand Ydj1's function, we have constructed and analyzed an array of Ydj1 mutants in vivo . Both the glucocorticoid receptor and the estrogen receptor exhibited elevated activity in the absence of hormone in all ydj1 mutant strains, indicating a strict requirement for Ydj1 activity in hormonal control . Glucocorticoid receptor containing a mutation in the carboxy-terminal transcriptional activation domain, AF-2, retained elevated basal activity, while mutation of the amino-terminal transactivation domain, AF-1, eliminated the elevated basal activity observed in ydj1 mutant strains . This result indicates that the source of activity is AF-1, which is normally repressed by the carboxy-terminal hormone binding domain in the absence of hormone . Chimeric proteins containing the hormone binding domain of the estrogen or glucocorticoid receptor fused to heterologous activation and DNA binding domains also exhibited elevated activity in the absence of hormone . Thus, Ydj1 mutants appear to increase basal receptor activity by altering the ability of the hormone binding domain of the receptor to repress nearby activation domains . We propose that Ydj1 functions to present steroid receptors to the Hsp90 pathway for folding and hormonal control . In the presence of Ydj1 mutants that fail to bind substrate efficiently, some receptor escapes the Hsp90 pathway, resulting in constitutive activity.

Mol Cell Biol, 2000 May, 20(9), 2984 - 95
A novel cold-sensitive allele of the rate-limiting enzyme of fatty acid synthesis, acetyl coenzyme A carboxylase, affects the morphology of the yeast vacuole through acylation of Vac8p; Schneiter R et al.; The yeast vacuole functions both as a degradative organelle and as a storage depot for small molecules and ions . Vacuoles are dynamic reticular structures that appear to alternately fuse and fragment as a function of growth stage and environment . Vac8p, an armadillo repeat-containing protein, has previously been shown to function both in vacuolar inheritance and in protein targeting from the cytoplasm to the vacuole . Both myristoylation and palmitoylation of Vac8p are required for its efficient localization to the vacuolar membrane (Y.-X . Wang, N . L . Catlett, and L . S . Weisman, J . Cell Biol . 140:1063-1074, 1998) . We report that mutants with conditional defects in the rate-limiting enzyme of fatty acid synthesis, acetyl coenzyme A carboxylase (ACC1), display unusually multilobed vacuoles, similar to those observed in vac8 mutant cells . This vacuolar phenotype of acc1 mutant cells was shown biochemically to be accompanied by a reduced acylation of Vac8p which was alleviated by fatty acid supplementation . Consistent with the proposed defect of acc1 mutant cells in acylation of Vac8p, vacuolar membrane localization of Vac8p was impaired upon shifting acc1 mutant cells to nonpermissive condition . The function of Vac8p in protein targeting, on the other hand, was not affected under these conditions . These observations link fatty acid synthesis and availability to direct morphological alterations of an organellar membrane.

Nucleic Acids Res, 2000 May 1, 28(9), 1913 - 20
Intramolecular interaction of yeast TFIIB in transcription control; Zhang DY et al.; The general transcription factor TFIIB is a key component in the eukaryotic RNA polymerase II (RNAPII) transcriptional machinery . We have previously shown that a yeast TFIIB mutant (called YR1m4) with four amino acid residues in a species-specific region changed to corresponding human residues affects the expression of genes activated by different activators in vivo . We report here that YR1m4 can interact with several affected activators in vitro . In addition, YR1m4 and other mutants with amino acid alterations within the same region can interact with TATA-binding protein (TBP) and RNAPII normally . However, YR1m4 is defective in supporting activator-independent transcription in assays con-ducted both in vitro and in vivo . We further demonstrate that the interaction between the C-terminal core domain and the N-terminal region is weakened in YR1m4 and other related TFIIB mutants . These results suggest that the intramolecular interaction property of yeast TFIIB plays an important role in transcription regulation in cells.

FEMS Microbiol Lett, 2000 Apr 15, 185(2), 147 - 50
Identification of a 49-kDa hydrophobic cell wall mannoprotein present in velum yeast which may be implicated in velum formation; Alexandre H et al.; Analysis of velum-forming yeast cell wall components released by beta-1,3-glucanase treatment were compared with those of a non velum-forming yeast . SDS-PAGE electrophoresis and Western blotting with ConA-peroxidase staining of mannoproteins allowed us to identify a 49-kDa mannoprotein present in the cell wall of the velum-forming yeast and hardly visible in the control . The cell wall nature of this protein was confirmed by labelling with the non-permeable sulfosuccinimydiyl-6-(biotinamido)hexanoate reagent . A partial purification of this mannoprotein by anion exchange HPLC followed by surface hydrophobicity determination revealed that the fraction containing the 49-kDa mannoprotein was the most hydrophobic . Since cell surface hydrophobicity plays an important role in aggregate formation, it is likely that this mannoprotein is involved in velum formation.

Curr Opin Genet Dev, 2000 Apr, 10(2), 169 - 77
Telomere transitions in yeast: the end of the chromosome as we know it; Cooper JP; Telomere functions vary as the cell cycle progresses . Recent results highlight fluctuating associations between telomeres and DNA polymerases, DNA-damage repair proteins, and centrosome components . These associations reflect diverse roles of telomeres in chromosome maintenance and in the orchestration of chromosome movements during meiosis.

J Biol Chem, 2000 Jun 16, 275(24), 18011 - 21
Premature structural changes at replication origins in a yeast minichromosome maintenance (MCM) mutant; Geraghty DS et al.; The Cdc7p protein kinase in the budding yeast Saccharomyces cerevisiae is thought to help trigger DNA replication by modifying one or more of the factors that assemble at replication origins (ARSs) . To investigate events catalyzed by Cdc7p, we compared the structure of replication origins in cells containing conditional mutations in Cdc7p and Cdc8p, a thymidylate kinase that is required for DNA synthesis . High resolution genomic footprinting indicated that the presumptive lagging strand template in ARS1 became highly sensitive to KMnO(4) modification after the CDC7 execution point . These results suggested that Cdc7p triggers DNA unwinding . The transition from late G(1) phase to the CDC7 execution point and from the CDC7 to the CDC8 execution points was accompanied by small but ARS-dependent changes in DNA topology . These results suggested that DNA unwinding before the CDC8 execution point either is highly localized or that the torsional stress associated with initial DNA unwinding is minimized by compensatory protein-DNA structural changes . The ARS DNA structural attributes evident in cells blocked at the CDC8 execution point were also evident in alpha-factor-blocked, G(1) phase cells containing the CDC7 bypass mutant mcm5/cdc46-bob1 . This result strongly suggests that the structural changes during the transition from the CDC7 to CDC8 execution points depend on the Cdc7p protein kinase and involve alteration of the minichromosome maintenance protein complex.

J Biol Chem, 2000 Jun 9, 275(23), 17391 - 8
Identification of a yeast transcription factor IID subunit, TSG2/TAF48; Reese JC et al.; The RNA polymerase II general transcription factor TFIID is a complex containing the TATA-binding protein (TBP) and associated factors (TAFs) . We have used a mutant allele of the gene encoding yeast TAF(II)68/61p to analyze its function in vivo . We provide biochemical and genetic evidence that the C-terminal alpha-helix of TAF(II)68/61p is required for its direct interaction with TBP, the stable incorporation of TBP into the TFIID complex, the integrity of the TFIID complex, and the transcription of most genes in vivo . This is the first evidence that a yeast TAF(II) other than TAF(II)145/130 interacts with TBP, and the implications of this on the interpretation of data obtained studying TAF(II) mutants in vivo are discussed . We have identified a high copy suppressor of the TAF68/61 mutation, TSG2, that has sequence similarity to a region of the SAGA subunit Ada1 . We demonstrate that it directly interacts with TAF(II)68/61p in vitro, is a component of TFIID, is required for the stability of the complex in vivo, and is necessary for the transcription of many yeast genes . On the basis of these functions, we propose that Tsg2/TAF(II)48p is the histone 2A-like dimerization partner for the histone 2B-like TAF(II)68/61p in the yeast TFIID complex.

Mol Biol Cell, 2000 Apr, 11(4), 1293 - 304
Mitotic chromosome condensation requires Brn1p, the yeast homologue of Barren; Lavoie BD et al.; In vitro studies suggest that the Barren protein may function as an activator of DNA topoisomerase II and/or as a component of the Xenopus condensin complex . To better understand the role of Barren in vivo, we generated conditional alleles of the structural gene for Barren (BRN1) in Saccharomyces cerevisiae . We show that Barren is an essential protein required for chromosome condensation in vivo and that it is likely to function as an intrinsic component of the yeast condensation machinery . Consistent with this view, we show that Barren performs an essential function during a period of the cell cycle when chromosome condensation is established and maintained . In contrast, Barren does not serve as an essential activator of DNA topoisomerase II in vivo . Finally, brn1 mutants display additional phenotypes such as stretched chromosomes, aberrant anaphase spindles, and the accumulation of cells with >2C DNA content, suggesting that Barren function influences multiple aspects of chromosome transmission and dynamics.

Mol Biol Cell, 2000 Apr, 11(4), 1197 - 211
Time-lapse video microscopy analysis reveals astral microtubule detachment in the yeast spindle pole mutant cnm67; Hoepfner D et al.; Saccharomyces cerevisiae cnm67Delta cells lack the spindle pole body (SPB) outer plaque, the main attachment site for astral (cytoplasmic) microtubules, leading to frequent nuclear segregation failure . We monitored dynamics of green fluorescent protein-labeled nuclei and microtubules over several cell cycles . Early nuclear migration steps such as nuclear positioning and spindle orientation were slightly affected, but late phases such as rapid oscillations and insertion of the anaphase nucleus into the bud neck were mostly absent . Analyzes of microtubule dynamics revealed normal behavior of the nuclear spindle but frequent detachment of astral microtubules after SPB separation . Concomitantly, Spc72 protein, the cytoplasmic anchor for the gamma-tubulin complex, was partially lost from the SPB region with dynamics similar to those observed for microtubules . We postulate that in cnm67Delta cells Spc72-gamma-tubulin complex-capped astral microtubules are released from the half-bridge upon SPB separation but fail to be anchored to the cytoplasmic side of the SPB because of the absence of an outer plaque . However, successful nuclear segregation in cnm67Delta cells can still be achieved by elongation forces of spindles that were correctly oriented before astral microtubule detachment by action of Kip3/Kar3 motors . Interestingly, the first nuclear segregation in newborn diploid cells never fails, even though astral microtubule detachment occurs.

Mol Biol Cell, 2000 Apr, 11(4), 1113 - 27
Functional interactions between the p35 subunit of the Arp2/3 complex and calmodulin in yeast; Schaerer-Brodbeck C et al.; The end9-1 (arc35-1) mutant was identified as an endocytosis mutant and is a mutant allele of ARC35 that encodes a subunit of the Arp2/3 complex . As for other mutants in the Arp2/3 complex, arc35-1 is defective for endocytosis and organization of the actin cytoskeleton . Both defects can be suppressed by overexpression of calmodulin . Analysis of a collection of temperature-sensitive cmd1 mutants for their ability to suppress either the endocytic defect and/or the actin defect indicates that the two defects are tightly coupled . We demonstrate that Arc35p and Cmd1p interact and that Arc35p is required for cortical localization of calmodulin . This is the first report linking Arp2/3 complex function with calmodulin through which it exercises at least one of its endocytic functions.

J Biol Chem, 2000 Jul 7, 275(27), 20814 - 21
The high resolution crystal structure of yeast hexokinase PII with the correct primary sequence provides new insights into its mechanism of action; Kuser PR et al.; Hexokinase is the first enzyme in the glycolytic pathway, catalyzing the transfer of a phosphoryl group from ATP to glucose to form glucose 6-phosphate and ADP . Two yeast hexokinase isozymes are known, namely PI and PII . The crystal structure of yeast hexokinase PII from Saccharomyces cerevisiae without substrate or competitive inhibitor is determined and refined in a tetragonal crystal form at 2.2-A resolution . The folding of the peptide chain is very similar to that of Schistosoma mansoni and previous yeast hexokinase models despite only 30% sequence identity between them . Distinct differences in conformation are found that account for the absence of glucose in the binding site . Comparison of the current model with S . mansoni and yeast hexokinase PI structures both complexed with glucose shows in atomic detail the rigid body domain closure and specific loop movements as glucose binds . A hydrophobic channel formed by strictly conserved hydrophobic residues in the small domain of the hexokinase is identified . The channel's mouth is close to the active site and passes through the small domain to its surface . The possible role of the observed channel in proton transfer is discussed.

J Biol Chem, 2000 Jun 16, 275(24), 18462 - 9
Functional analysis of Plp1 and Plp2, two homologues of phosducin in yeast; Flanary PL et al.; Mammalian phosducins are known to bind G protein betagamma subunits in vitro, and are postulated to regulate their signaling function in vivo . Here we describe two homologues of phosducin in yeast, called PLP1 and PLP2 . Both gene products were cloned, expressed, and purified as glutathione S-transferase fusions . Of the two isoforms, Plp1 bound most preferentially to Gbetagamma . Binding was enhanced by pheromone stimulation and by the addition of GTPgammaS, conditions that favor dissociation of Gbetagamma from Galpha . Gene disruption mutants and gene overexpression plasmids were prepared and analyzed for changes in signaling and nonsignaling phenotypes . Haploid spore products bearing the plp2Delta mutant failed to grow, suggesting that PLP2 is an essential gene . Cell viability was not restored by a mutation in STE7 that blocks signaling downstream of the G protein . Haploid products bearing the plp1Delta mutant were viable and exhibited a 6-7% increase in pheromone-mediated gene induction . Cells overexpressing PLP1 or PLP2 exhibited a 70-80% decrease in gene induction but no change in pheromone-mediated growth arrest . These data indicate that phosducin can selectively regulate early signaling events following pheromone stimulation and has an essential role in cell growth independent of its regulatory role in cell signaling.

J Biol Chem, 2000 Jun 9, 275(23), 17900 - 8
Sfb2p, a yeast protein related to Sec24p, can function as a constituent of COPII coats required for vesicle budding from the endoplasmic reticulum; Higashio H et al.; The COPII coat is required for vesicle budding from the endoplasmic reticulum (ER), and consists of two heterodimeric subcomplexes, Sec23p/Sec24p, Sec13p/Sec31p, and a small GTPase, Sar1p . We characterized a yeast mutant, anu1 (abnormal nuclear morphology) exhibiting proliferated ER as well as abnormal nuclear morphology at the restrictive temperature . Based on the finding that ANU1 is identical to SEC24, we confirmed a temperature-sensitive protein transport from the ER to the Golgi in anu1-1/sec24-20 cells . Overexpression of SFB2, a SEC24 homologue with 56% identity, partially suppressed not only the mutant phenotype of sec24-20 cells but also rescued the SEC24-disrupted cells . Moreover, the yeast two-hybrid assay revealed that Sfb2p, similarly to Sec24p, interacted with Sec23p . In SEC24-disrupted cells rescued by overexpression of SFB2, some cargo proteins were still retained in the ER, while most of the protein transport was restored . Together, these findings strongly suggest that Sfb2p functions as the component of COPII coats in place of Sec24p, and raise the possibility that each member of the SEC24 family of proteins participates directly and/or indirectly in cargo-recognition events with its own cargo specificity at forming ER-derived vesicles.

Biochem J, 2000 Apr 15, 347(Pt 2), 363 - 8
Yeast cell-free system that catalyses joint-molecule formation in a Rad51p- and Rad52p-dependent fashion; Nagaraj V et al.; One of the central reactions of homologous recombination is the invasion of a single strand of DNA into a homologous duplex to form a joint molecule . Here we describe the isolation of a cell-free system from meiotic yeast cells that catalyses joint-molecule formation in vitro . The active components in the system required ATP and homologous DNA and operated in both 0.5 and 13 mM MgCl(2) . When the cell-free system was prepared from rad51/rad51 and rad52/rad52 mutants and joint-molecule formation was assayed at 0.5 mM MgCl(2), the specific activity decreased to 6% and 13.8% respectively of the wild-type level . However, when the same mutant extracts were premixed, joint-molecule formation increased 4-8-fold, i.e . the mutant extracts exhibited complementation in vitro . These results demonstrated that Rad51p and Rad52p were required for optimal joint-molecule formation at 0.5 mM MgCl(2) . Intriguingly, however, Rad51p and Rad52p seemed to be more dispensable at higher concentrations of MgCl(2) (13 mM) . Further purification of the responsible activity has proven problematical, but it did flow through a sizing column as a single peak (molecular mass 1.2 MDa) that was co-eluted with Rad51p and RFA, the eukaryotic single-stranded DNA-binding protein . All of these characteristics are consistent with the known properties of the reaction in vivo and suggest that the new cell-free system will be suitable for purifying enzymes involved in homologous recombination.

RNA, 2002 Aug, 8(8), 1068 - 77
How Slu7 and Prp18 cooperate in the second step of yeast pre-mRNA splicing; James SA et al.; Slu7 and Prp18 act in concert during the second step of yeast pre-mRNA splicing . Here we show that the 382-amino-acid Slu7 protein contains two functionally important domains: a zinc knuckle (122CRNCGEAGHKEKDC135) and a Prp18-interaction domain (215EIELMKLELY224) . Alanine cluster mutations of 215EIE217 and 221LELY224 abrogated Slu7 binding to Prp18 in a two-hybrid assay and in vitro, and elicited temperature-sensitive growth phenotypes in vivo . Yet, the mutations had no impact on Slu7 function in pre-mRNA splicing in vitro . Single alanine mutations of zinc knuckle residues Cys122, His130, and Cys135 had no effect on cell growth, but caused Slu7 function during pre-mRNA splicing in vitro to become dependent on Prp18 . Specifically, zinc knuckle mutants required Prp18 in order to bind to the spliceosome . Compound mutations in both Slu7 domains (e.g., C122A-EIE, H130A-EIE, and C135A-EIE) were lethal in vivo and abolished splicing in vitro, suggesting that the physical interaction between Slu7 and Prp18 is important for cooperation in splicing . Depletion/reconstitution studies coupled with immunoprecipitations suggest that second step factors are recruited to the spliceosome in the following order: Slu7 --> Prp18 --> Prp22 . All three proteins are released from the spliceosome after step 2 concomitant with release of mature mRNA.

RNA, 2002 Aug, 8(8), 1011 - 33
The 5' and 3' domains of yeast U6 snRNA: Lsm proteins facilitate binding of Prp24 protein to the U6 telestem region; Ryan DE et al.; The 5' and 3' domains of yeast U6 snRNA contain sequences that are thought to be important for binding to Prp24 and Lsm proteins . By extensive mutational analysis of yeast U6 snRNA, we confirmed that the 3' terminal uridine tract of U6 snRNA is important for U6 binding to Lsm proteins in yeast . Binding of Prp24 protein to U6 RNA is dependent on or is strongly enhanced by U6 binding of Lsm proteins . This supports a model for U6 snRNP assembly in which U6 RNA binds to the Lsm2-8 core prior to binding Prp24 protein . Using compensatory base-pairing analysis, we show that at least half of the recently identified U6 telestem as well as a nucleotide sequence in the other half of the telestem are important for binding of U6 RNA to Prp24 protein . Surprisingly, disruption of base pairing in the unconfirmed half of the telestem enhanced U6-Prp24 binding . Truncation of the entire 3' terminal domain or nearly the entire 5' terminal domain of yeast U6 allowed for detectable levels of splicing to proceed in vitro . In addition to gaining knowledge of the function of the 5' and 3' domains of yeast U6, our results help define the minimal set of requirements for yeast U6 RNA function in splicing . We present a revised secondary structural model of yeast U6 snRNA in free U6 snRNPs.

Nat Rev Genet, 2002 Sep, 3(9), 653 - 61
Treasures and traps in genome-wide data sets: case examples from yeast; Grunenfelder B et al.; Since the publication of the Saccharomyces cerevisiae genome sequence, much effort has been dedicated to developing high-throughput techniques to generate comprehensive information about the function and dynamics of all genes in this yeast's genome . These techniques have generated data sets that typically contain large amounts of reliable and valuable biological information . Nevertheless, there are also uncertainties that are associated with such large-scale studies, which we discuss in this review . These uncertainties increase with the complexity of the organism under study . On the basis of the results from yeast, we should learn much from human and mouse genomic data sets . However, as with yeast data sets, they might also contain misleading results.

Biochem Biophys Res Commun, 2002 Sep 6, 296(5), 1310 - 6
Inhibition of yeast ribosomal stalk phosphorylation by Cu-Zn superoxide dismutase; Zielinski R et al.; Reversible phosphorylation of acidic ribosomal proteins of Saccharomyces cerevisiae is an important mechanism, regulating the number of active ribosomes . The key role in regulation of this process is played by specific, second messenger-independent protein kinases . A new protein-inhibitor regulating activity of PK60S kinase has been purified from yeast extracts and characterised . Peptide mass fingerprinting (PMF) and amino-acid sequence analysis by Post Source Decay (PSD) have identified the inhibitor as a Cu-Zn superoxide dismutase (SOD) . Inhibition by SOD is competitive with respect to protein substrates-P proteins and 80S ribosome-with K(i) values of 3.7 microM for P2A protein and 0.6 microM for 80S ribosomes . A close correlation was found between the state of phosphorylation of P proteins in diauxic shift and logarithmic growth yeast cells and activity of SOD . The possible mechanism of regulation of PK60S activity, and participation of SOD protein in regulation of 80S-ribosome activity in stress conditions, is discussed.

Biochem Biophys Res Commun, 2002 Sep 6, 296(5), 1245 - 50
A beta-tubulin-like cDNA expressed specifically in elongating cotton fibers induces longitudinal growth of fission yeast; Ji S et al.; Using cDNA Representational Difference Analysis (RDA) techniques, we isolated a cDNA that was expressed specifically in cotton fibers but not in the ovules of a fuzzless-lintless mutant (fl) . We designated it as Gh-BTubL for it shares high sequence identity with known plant and yeast beta-tubulins . RT-PCR and robotic cDNA dot blot analyses indicated that the expression of Gh-BTubL was correlated with the elongation pattern of cotton fibers . In situ hybridization results verified that there was no Gh-BTubL mRNA in fl ovules while it was easily detected in the elongating wild type cotton fiber cells . Overexpression of Gh-BTubL in fission yeast induced longitudinal growth of the host cells by 1.74-fold, with no apparent effect on other aspects of the host cells . We suggest that Gh-BTubL plays an important role in cotton fiber elongation and we believe that elucidation of the control mechanisms for expression of tubulin-like proteins may help improve fiber quality and productivity.

Carbohydr Res, 2002 Sep 3, 337(15), 1367 - 71
Synthesis of a mannose heptasaccharide of the pathogenic yeast, Candida glabrata IFO 0622 strain; Zeng Y et al.; An effective synthesis of the mannose heptasaccharide existing in the pathogenic yeast, Candida glabrata IFO 0622 strain was achieved via TMSOTf-promoted condensation of a tetrasaccharide donor 13 with a trisaccharide acceptor 16, followed by deprotection . The tetrasaccharide 13 was constructed by coupling of 2,3,4,6-tetra-O-benzoyl-alpha-D-mannopyranosyl-(1-->3)-2,4,6-tri-O-acetyl-alpha-D-mannopyranosyl trichloroacetimidate (7) with allyl 3,4,6-tri-O-benzoyl-alpha-D-mannopyranosyl-(1-->2)-3,4,6-tri-O-benzoyl-alpha-D-mannopyranoside (10), followed by deallylation and trichloroacetimadation . The trisaccharide 16 was obtained by coupling of 6-O-acetyl-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl trichloroacetimidate with 10, and subsequent 6-O-deacetylation . The disaccharide 7 was prepared through coupling of perbenzoylated mannosyl trichloroacetimidate with 4,6-O-benzylidene-1,2-O-ethylidene-beta-D-mannopyranose, then simultaneous debenzylidenation and deethylidenation, and subsequent acetylation, selective 1-O-deacetylation, and trichloroacetimidation . The disaccharide 10 was obtained by self-condensation of 3,4,6-tri-O-benzoyl-1,2-O-allyloxyethylidene-beta-D-mannopyranose, followed by selective 2-O-deacetylation.

FEMS Microbiol Lett, 2002 Aug 27, 214(1), 101 - 6
Identifying Pex21p as a protein that specifically interacts with yeast seryl-tRNA synthetase; Rocak S et al.; The interaction of Saccharomyces cerevisiae seryl-tRNA synthetase (SerRS) with peroxin Pex21p was identified in a two-hybrid screen with SerRS as bait . This was confirmed by an in vitro binding assay with truncated Pex21p fused to glutathione S-transferase . Furthermore, purified Pex21p acts as an activator of yeast seryl-tRNA synthetase in aminoacylation in vitro, revealing the functional significance of the Pex21p-SerRS interaction . Pex21p is a protein involved in the peroxisome biogenesis {Purdue, P.E., Yang, X . and Lazarow, P.B., J . Cell Biol . 143 (1998) 1859-1869} . Since eukaryotic aminoacyl-tRNA synthetases are known to participate in assembles with other synthetases and non-synthetase proteins, we propose that this unusual interaction reflects another function of the peroxin.

Phytother Res, 2002 Aug, 16(5), 414 - 6
Estrogenic activity of Nigella damascena extracts, evaluated using a recombinant yeast screen; Agradi E et al.; We used the yeast estrogen screen (YES) containing a human estrogen receptor to evaluate the estrogenic activity of extracts obtained from Nigella damascena seeds . Alcohol extracts obtained by direct extraction of seeds showed a low estrogenic activity, while the alcohol extract obtained after extraction with solvents of increasing polarity showed a strong estrogenic activity . This suggests the presence in Nigella of polar components whose activity can be clearly demonstrated after previous elimination of interacting apolar components that may mask the activity of more polar components . The response of both alcohol fractions follow a bell-shaped curve indicating a concentration-dependent relationship .

J Biol Chem, 2002 Nov 15, 277(46), 44202 - 7 Epub 2002 Aug 27.
A complex of the Srb8, -9, -10, and -11 transcriptional regulatory proteins from yeast; Borggrefe T et al.; The Srb8, -9, -10, and -11 proteins of yeast have been isolated as a discrete, stoichiometric complex . The isolated complex phosphorylates the C-terminal domain (CTD) of the largest subunit of RNA polymerase II at serines 2 and 5 . In addition to the previously reported human homologs of Srb10 and 11, we have identified TRAP230/ARC240 and TRAP240/ARC250 as the human homologs of Srb8 and Srb9, showing the entire Srb8/9/10/11 complex is conserved from yeast to humans.

Biochem Biophys Res Commun, 2002 Aug 30, 296(4), 949 - 53
Supercomplex formation between Mlh1-Mlh3 and Sgs1-Top3 heterocomplexes in meiotic yeast cells; Wang TF et al.; The genome of Saccharomyces cerevisia encodes four mismatch repair MutL proteins and these proteins form three heterocomplexes: Mlh1-Mlh2, Mlh1-Mlh3, and Mlh1-Pms1 . Only, the Mlh1-Mlh3 heterocomplex has been implicated specifically in promotion of meiotic crossing-over . In this report, we show that yeast Mlh3 co-immunoprecipitates with Sgs1 helicase in sporulating cells at late stage of meiotic prophase I . Sgs1, a member of the RecQ DNA helicase family, appears to form a stable complex with topoisomerase III (Top3) during meiosis . We suggest that Mlh1-Mlh3 heterocomplex may act as a molecular matchmaker to coordinate Sgs1-Top3 complex in the resolution of meiotic recombination intermediates.

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi, 2002 Jun, 16(2), 154 - 6
{Transcriptional activation function of hepatitis B virus Pre S1 protein in yeast}; Xiao S et al.; BACKGROUND: To explore the feasibility of cloning of the hepatocyte receptor interacting with the Pre S1 protein of HBV by two hybrid system . METHODS: Yeast expression plasmids encoding fusion proteins of full length or portions of Pre S1 of HBV and DNA binding domain of yeast protein GAL4 were constructed and used to transform yeast reporter strain SFY526 . Reporter gene product ?galactosidase activity was assayed as a measure of transcription activation in yeast . Mammalian expression plasmid encoding fusion proteins of full length Pre S1 and DNA binding domain of GAL4 was constructed and used to cotransfect hepatoma cell line Huh?7 together with CAT reporter plasmid . Cell extracts were assayed for CAT activity by thin?layer chromatography . RESULTS: The fusion proteins of full length Pre S1 protein and GAL4 DNA binding domain present transcriptional activation function in yeast . The transcription activating sequence is localized to the 21 to 47 amino acids of Pre S1 protein Fusion proteins of full length Pre S 1 and GAL 4 DNA binding domain do not show transcriptional activation function in mammalian cells . CONCLUSION: The transcriptional activating sequence of HBVPre S1 protein in yeast overlaps the hepatocyte receptor binding site . The transcriptional activation function of HBV Pre S1 protein in yeast may prevent researchers?from using yeast two hybrid system to clone HBV receptor interacting with Pre S1 protein . However, the Pre S1 protein does not show transcriptional activation function in mammalian cells . Mammalian two?hybrid system may be a practical method to clone the HBV hepatocyte receptor interacting with Pre S1 protein.

Genetics, 2002 Aug, 161(4), 1465 - 82
Nuclear import of Upf3p is mediated by importin-alpha/-beta and export to the cytoplasm is required for a functional nonsense-mediated mRNA decay pathway in yeast; Shirley RL et al.; Upf3p, which is required for nonsense-mediated mRNA decay (NMD) in yeast, is primarily cytoplasmic but accumulates inside the nucleus when UPF3 is overexpressed or when upf3 mutations prevent nuclear export . Upf3p physically interacts with Srp1p (importin-alpha) . Upf3p fails to be imported into the nucleus in a temperature-sensitive srp1-31 strain, indicating that nuclear import is mediated by the importin-alpha/beta heterodimer . Nuclear export of Upf3p is mediated by a leucine-rich nuclear export sequence (NES-A), but export is not dependent on the Crm1p exportin . Mutations identified in NES-A prevent nuclear export and confer an Nmd(-) phenotype . The addition of a functional NES element to an export-defective upf(-) allele restores export and partially restores an Nmd(+) phenotype . Our findings support a model in which the movement of Upf3p between the nucleus and the cytoplasm is required for a fully functional NMD pathway . We also found that overexpression of Upf2p suppresses the Nmd(-) phenotype in mutant strains carrying nes-A alleles but has no effect on the localization of Upf3p . To explain these results, we suggest that the mutations in NES-A that impair nuclear export cause additional defects in the function of Upf3p that are not rectified by restoration of export alone.

Genetics, 2002 Aug, 161(4), 1453 - 64
Loss of ypk1 function causes rapamycin sensitivity, inhibition of translation initiation and synthetic lethality in 14-3-3-deficient yeast; Gelperin D et al.; 14-3-3 proteins bind to phosphorylated proteins and regulate a variety of cellular activities as effectors of serine/threonine phosphorylation . To define processes requiring 14-3-3 function in yeast, mutants with increased sensitivity to reduced 14-3-3 protein levels were identified by synthetic lethal screening . One mutation was found to be allelic to YPK1, which encodes a Ser/Thr protein kinase . Loss of Ypk function causes hypersensitivity to rapamycin, similar to 14-3-3 mutations and other mutations affecting the TOR signaling pathway in yeast . Similar to treatment with rapamycin, loss of Ypk function disrupted translation, at least in part by causing depletion of eIF4G, a central adaptor protein required for cap-dependent mRNA translation initiation . In addition, Ypk1 as well as eIF4G protein levels were rapidly depleted upon nitrogen starvation, but not during glucose starvation, even though both conditions inhibit translation initiation . These results suggest that Ypk regulates translation initiation in response to nutrient signals, either through the TOR pathway or in a functionally related pathway parallel to TOR.

Genetics, 2002 Aug, 161(4), 1425 - 35
A heritable structural alteration of the yeast mitochondrion; Lockshon D; Prions have revived interest in hereditary change that is due to change in cellular structure . How pervasive is structural inheritance and what are its mechanisms? Described here is the initial characterization of {Leu(P)}, a heritable structural change of the mitochondrion of Saccharomyces cerevisiae that often but not always accompanies the loss of all or part of the mitochondrial genome . Three phenotypes are reported in {Leu(P)} vs . {Leu(+)} strains: twofold slower growth, threefold slower growth in the absence of leucine, and a marked delocalization of nuclear-encoded protein destined for the mitochondrion . Introduction of mitochondria from a {Leu(+)} strain by cytoduction can convert a {Leu(P)} strain to {Leu(+)} and vice versa . Evidence against the Mendelian inheritance of the trait is presented . The incomplete dominance of {Leu(P)} and {Leu(+)} and the failure of HSP104 deletion to have any effect suggest that the trait is not specified by a prion but instead represents a new class of heritable structural change.

Neuro Endocrinol Lett, 2002 Aug, 23(4), 287 - 8
Mating types in yeast, vomeronasal organ in rodents, homosexuality in humans: does a guiding thread exist?
Oliva D.
Pheromones and their receptors are the molecules used by very different organisms in order to join two haploid cells . It happens evidently in yeast, since the two blending haploid cells are also the two mating organisms, whereas in rodents pheromone receptors are the triggers of the vomeronasal system which, supervising sexual behaviors, is responsible for copulation and therefore for fertilization . The debate is still open about the real significance of pheromones in humans but a working vomeronasal organ, able to recognize pheromones of the same sex, could be the simplest biological explanation of homosexuality . This hypothesis is discussed and connected with some well known experimental data.

J Biol Chem, 2002 Nov 1, 277(44), 41463 - 72 Epub 2002 Aug 22.
The extracellular N-terminal domain and transmembrane domains 1 and 2 mediate oligomerization of a yeast G protein-coupled receptor; Overton MC et al.; G protein-coupled receptors (GPCRs) can form homodimers/oligomers and/or heterodimers/oligomers . The mechanisms used to form specific GPCR oligomers are poorly understood because the domains that mediate such interactions and the step(s) in the secretory pathway where oligomerization occurs have not been well characterized . Here we have used subcellular fractionation and fluorescence resonance energy transfer (FRET) experiments to show that oligomerization of a GPCR (alpha-factor receptor; STE2 gene product) of the yeast Saccharomyces cerevisiae occurs in the endoplasmic reticulum . To identify domains of this receptor that mediate oligomerization, we used FRET and endocytosis assays of oligomerization in vivo to analyze receptor deletion mutants . A mutant lacking the N-terminal extracellular domain and transmembrane (TM) domain 1 was expressed at the cell surface but did not self-associate . In contrast, a receptor fragment containing only the N-terminal extracellular domain and TM1 could self-associate and heterodimerize with wild type receptors . Analysis of other mutants suggested that oligomerization is facilitated by the N-terminal extracellular domain and TM2 . Therefore, the N-terminal extracellular domain, TM1, and TM2 appear to stabilize alpha-factor receptor oligomers . These domains may form an interface in contact or domain-swapped oligomers . Similar domains may mediate dimerization of certain mammalian GPCRs.

Rev Med Chil, 2002 Jun, 130(6), 661 - 5
{Systemic yeast infections in a general hospital . Correlation between study of susceptibility in vitro and patient survival to the fungal infection episode}; Tapia C et al.; BACKGROUND: Fungi are important causal agents of nosocomial infections, that usually have high mortality rates . AIM: To evaluate the species distribution and susceptibility patterns of deep yeast infections in a General Hospital and to correlate those results with patient survival . MATERIAL AND METHODS: Twenty one strains (from five pediatric and 16 from adult patients) were studied . Antifungal Susceptibility Testing (AST) to Amphotericin B (Anfb), Fluorocytosine (5FC), Fluconazole (FZ) and Itraconazole (IZ) was performed according to the EUCAST document . Clinical data of patients was obtained and survival to the infection was recorded . RESULTS: C . albicans was isolated in 11 samples (52%), C . parapsilosis in three samples (14%), C . glabrata in two samples (9%), C . tropicalis in one sample (5%) and C . neoformans in four samples (19%) . Twenty three percent of fungi were recovered at the Surgical Intensive Care Unit . The MICs ranged between 0.25 and 0.5 microgram/mL for Anfb; between 0.25 and 16 micrograms/ml for SFC, between 0.12 and 32 micrograms/mL for FZ and 0.015 and 0.5 microgram/mL for IZ . No association between antifungal susceptibility and patient survival was observed . CONCLUSIONS: C . albicans continues to be the most frequently isolated yeast, however, non-albicans species are an emergent group causing nosocomial infections . Surgical procedures are the main source of fungal infections in this sample.

Radiat Prot Dosimetry, 2002, 99(1-4), 269 - 72
DNA double strand break induction in yeast; Kiefer J et al.; The induction of DNA double strand breaks (DSBs) by accelerated heavy ions was systematically measured in diploid yeast cells . Particles were provided by the accelerators at GSI, Darmstadt, and HMI, Berlin . DNA was separated using pulsed field gel electrophoresis and the intensity of the largest bands used to determine the loss of molecular weight . Since the DNA content of each chromosome is exactly known absolute values for DSB induction can be measured without calibration procedures . Ions used range from protons to uranium with LET values between 2 and about 15,000 keV.micron-1 . Induction cross sections increase in the lower LET region approaching a plateau around 200 keV.micron-1 . With higher LET values the dependence can no longer be described by a common curve with each ion showing a specific behaviour . With very heavy particles the influence of the penumbra becomes obvious: cross sections decrease with LET because of the reduced penumbra extensions . Classical target theory would predict cross sections to follow a simple saturation function which is not substantiated by the data . Track structure analysis as introduced by Butts and Katz in 1967 is also not able to predict the experimental results . A semi-empirical fit indicates a linear-quadratic dependence of induction cross sections on LET up to about 1000 keV.micron-1 . RBE for DSB induction rises above unity reaching a maximum of about 2.5 around 200 keV.micron-1 . This is different from many experiments in mammalian cells and is presumably due to differences in chromatin structure since yeast cells seem to lack a functional III histone.

Proc Natl Acad Sci U S A, 2002 Dec 10, 99 Suppl 4, 16438 - 45 Epub 2002 Aug 22.
Histone H3 lysine 4 methylation is mediated by Set1 and promotes maintenance of active chromatin states in fission yeast; Noma K et al.; Methylation of histone H3 at lysine 4 (H3 Lys-4) or lysine 9 (H3 Lys-9) is known to define active and silent chromosomal domains respectively from fission yeast to humans . However, in budding yeast, H3 Lys-4 methylation is also necessary for silent chromatin assembly at telomeres and ribosomal DNA . Here we demonstrate that deletion of set1, which encodes a protein containing an RNA recognition motif at its amino terminus and a SET domain at the carboxy terminus, abolishes H3 Lys-4 methylation in fission yeast . Unlike in budding yeast, Set1-mediated H3 Lys-4 methylation is not required for heterochromatin assembly at the silent mating-type region and centromeres in fission yeast . Our analysis suggests that H3 Lys-4 methylation is a stable histone modification present throughout the cell cycle, including mitosis . The loss of H3 Lys-4 methylation in set1Delta cells is correlated with a decrease in histone H3 acetylation levels, suggesting a mechanistic link between H3 Lys-4 methylation and acetylation of the H3 tail . We suggest that methylation of H3 Lys-4 primarily acts in the maintenance of transcriptionally poised euchromatic domains, and that this modification is dispensable for heterochromatin formation in fission yeast, which instead utilizes H3 Lys-9 methylation.

Funct Integr Genomics, 2002 Sep, 2(4-5), 212 - 37 Epub 2002 May 24.
Yeast genomic databases and the challenge of the post-genomic era; Garrels JI; Since the completion of the yeast genome sequence in 1996, three genomic databases, the Saccharomyces Genome Database, the Yeast Proteome Database, and MIPS (produced by the Munich Information Center for Protein Sequences), have organized published knowledge of yeast genes and proteins onto the framework of the genome . Now, post-genomic technologies are producing large-scale datasets of many types, and these pose new challenges for knowledge integration . This review first examines the structure and content of the three genomic databases, and then draws from them and other resources to examine the ways knowledge from the literature, genome, and post-genomic experiments is stored, integrated, and disseminated . To better understand the impact of post-genomic technologies, 20 collections of post-genomic data were analyzed relative to a set of 243 previously uncharacterized genes . The results indicate that post-genomic technologies are providing rich new information for nearly all yeast genes, but data from these experiments is scattered across many Web sites and the results from these experiments are poorly integrated with other forms of yeast knowledge . Goals for the next generation of databases are set forth which could lead to better access to yeast knowledge for yeast researchers and the entire scientific community.

Funct Integr Genomics, 2002 Sep, 2(4-5), 181 - 92 Epub 2002 Apr 30.
The genomics of yeast responses to environmental stress and starvation; Gasch AP et al.; Unicellular organisms such as yeast have evolved to survive constant fluctuations in their external surroundings by rapidly adapting their internal systems to meet the challenges of each new environment . One aspect of this cellular adaptation is the reorganization of genomic expression to the program required for growth in each environment . The reprogramming of genomic expression can be unveiled using DNA microarrays, which measure the relative transcript abundance of essentially every gene in an organism's genome . Characterizing environmentally triggered gene expression changes provides insights into when, where, and how each gene is expressed and offers a glimpse at the physiological response of the cells to changes in their surroundings . This review will focus on the genomic expression responses of the budding yeast Saccharomyces cerevisiae to diverse environmental changes, highlighting some of the themes that have emerged from the collection of published yeast genomic expression studies . The results of these studies present insights as to how yeast cells sense and respond to each new environment, and suggest mechanisms that this organism uses to survive stressful environmental changes.

Funct Integr Genomics, 2002 Sep, 2(4-5), 171 - 80 Epub 2002 Jul 10.
Global analysis of gene expression in yeast; Horak CE et al.; In the past decade, there has been an intense effort to comprehensively catalogue the expressed genes in the yeast Saccharomyces cerevisiae and to determine the absolute and relative abundance of transcript and protein levels under different cellular conditions . Several methods have been developed to monitor gene expression: DNA microarray analysis, Serial Analysis of Gene Expression (SAGE), kinetic RT-PCR and monitoring expression of beta-galactosidase fusion proteins . These techniques have been used to measure transcript and protein abundance in different developmental states and under different environmental stimuli . A wealth of expression data for yeast is now publicly available through several web sites . The expression information that exists has the obvious benefits of providing a better understanding of the gene expression patterns that accompany changes in a yeast cell's environmental and developmental states . This data has also, however, provided clues to unraveling the complicated questions surrounding gene regulation: why and how is gene expression controlled?

Mol Cell Biol, 2002 Sep, 22(18), 6480 - 6
Stimulation of 3'-->5' exonuclease and 3'-phosphodiesterase activities of yeast apn2 by proliferating cell nuclear antigen; Unk I et al.; The Apn2 protein of Saccharomyces cerevisiae contains 3'-->5' exonuclease and 3'-phosphodiesterase activities, and these activities function in the repair of DNA strand breaks that have 3'-damaged termini and which are formed in DNA by the action of oxygen-free radicals . Apn2 also has an AP endonuclease activity and functions in the removal of abasic sites from DNA . Here, we provide evidence for the physical and functional interaction of Apn2 with proliferating cell nuclear antigen (PCNA) . As indicated by gel filtration and two-hybrid studies, Apn2 interacts with PCNA both in vitro and in vivo and mutations in the consensus PCNA-binding motif of Apn2 abolish this interaction . Importantly, PCNA stimulates the 3'-->5' exonuclease and 3'-phosphodiesterase activities of Apn2 . We have examined the involvement of the interdomain connector loop (IDCL) and of the carboxy-terminal domain of PCNA in Apn2 binding and found that Apn2 binds PCNA via distinct domains dependent upon whether the binding is in the absence or presence of DNA . In the absence of DNA, Apn2 binds PCNA through its IDCL domain, whereas in the presence of DNA, when PCNA has been loaded onto the template-primer junction by replication factor C, the C-terminal domain of PCNA mediates the binding.

Mol Cell Biol, 2002 Sep, 22(18), 6406 - 16
Targeted histone acetylation at the yeast CUP1 promoter requires the transcriptional activator, the TATA boxes, and the putative histone acetylase encoded by SPT10; Shen CH et al.; The relationship between chromatin remodeling and histone acetylation at the yeast CUP1 gene was addressed . CUP1 encodes a metallothionein required for cell growth at high copper concentrations . Induction of CUP1 with copper resulted in targeted acetylation of both H3 and H4 at the CUP1 promoter . Nucleosomes containing upstream activating sequences and sequences farther upstream were the targets for H3 acetylation . Targeted acetylation of H3 and H4 required the transcriptional activator (Ace1p) and the TATA boxes, suggesting that targeted acetylation occurs when TATA-binding protein binds to the TATA box or at a later stage in initiation . We have shown previously that induction results in nucleosome repositioning over the entire CUP1 gene, which requires Ace1p but not the TATA boxes . Therefore, the movement of nucleosomes occurring on CUP1 induction is independent of targeted acetylation . Targeted acetylation of both H3 and H4 also required the product of the SPT10 gene, which encodes a putative histone acetylase implicated in regulation at core promoters . Disruption of SPT10 was lethal at high copper concentrations and correlated with slower induction and reduced maximum levels of CUP1 mRNA . These observations constitute evidence for a novel mechanism of chromatin activation at CUP1, with a major role for the TATA box.

J Biol Chem, 2002 Oct 25, 277(43), 41101 - 9 Epub 2002 Aug 20.
GTP-yeast actin; Wen KK et al.; Because of the apparently greater conformational flexibility of yeast versus muscle actin and the ability of other members in the actin protein superfamily to efficiently use both ATP and GTP, we assessed the ability of yeast actin to function with GTP . Etheno-ATP exchange studies showed that the binding of GTP to yeast actin is about 1/9 as tight as that of ATP in contrast to the 1/1,240 ratio for muscle actin . Proteolysis of GTP-bound G-yeast actin suggests that the conformation of subdomain 2 is very much like that of ATP-bound actin, but CD studies show that GTP-bound actin is less thermostable than ATP-bound actin . GTP-actin polymerizes with an apparent critical concentration of 1.5 microm, higher than that of ATP-actin (0.3 microm) although filament structures observed by electron microscopy were similar . Yeast actin hydrolyzes GTP in a polymerization-dependent manner, and GTP-bound F-actin decorates with the myosin S1 . Conversion of Phe(306) in the nucleotide binding site to the Tyr found in muscle actin raised the nucleotide discrimination ratio from the 1/9 of wild-type actin to 1/125 . This result agrees with modeling that predicts that removal of the Tyr hydroxyl will create a space for the C2 amino group of the GTP guanine.

Proc Natl Acad Sci U S A, 2002 Sep 3, 99(18), 11622 - 7 Epub 2002 Aug 19.
SALSA, a variant of yeast SAGA, contains truncated Spt7, which correlates with activated transcription; Sterner DE et al.; Spt-Ada-Gcn5 acetyltransferase (SAGA) is a previously described histone acetyltransferase/transcriptional coactivator complex in yeast . At promoters of certain genes (HIS3 and TRP3), SAGA has an inhibitory function involving a nonproductive TATA-binding protein interaction mediated by the Spt3 and Spt8 subunits . Related to this, Spt8-less SAGA is a major form of the complex under activating conditions for these genes . In the present study, we purify this activation-specific complex, called SALSA (SAGA altered, Spt8 absent) . Besides lacking Spt8, SALSA contains Spt7 subunit that is truncated . Examining the role of this subunit, we find that C-terminally truncated SPT7 resulted in derepressed HIS3 transcription . Furthermore, when grown in rich media (repressing conditions), wild-type cells yielded predominantly SAGA, but Spt7 C-terminal truncations resulted primarily in a form of complex similar to SALSA . Thus, SALSA-like structure and activating function can be partially recapitulated in yeast by truncating the C terminus of Spt7 . Overall, these results lead to a model that for a subset of promoters SAGA is inhibitory through Spt3, Spt8, and an Spt8-interacting subdomain of Spt7, whereas SALSA is a form of complex for positive transcriptional regulation . These data clarify a mechanism by which a transcriptional regulatory complex can switch between positive and negative modulation.

In a process that superficially resembles dimorphism, Candida albicans undergoes a process called "phenotypic switching", in which different cellular morphologies are generated spontaneously. One of the classically studied strains that undergoes phenotypic switching is WO-1, which consists of two phases - one that grows as smooth white colonies and one that is rod-like and grows as flat gray colonies. The other strain known to undergo switching is 3153A; this strain produces at least seven different colony morphologies. In both the WO-1 and 3153A strains, the different phases convert spontaneously to the other(s) at a low frequency. The switching is reversible, and colony type can be inherited from one generation to another. While several genes that are expressed differently in different colony morphologies have been identified, some recent efforts have focussed on what might be controlling these changes. Further, whether there is a potential molecular link between dimorphism and phenotypic switching is a tantalizing question.

In the 3153A strain, a gene called SIR2 (for silent information regulator) has been found that seems to be important for phenotypic switching. SIR2 was originally found in Saccharomyces cerevisiae (brewer's yeast), where it is involved in chromosomal silencing - a form of transcriptional regulation in which regions of the genome are reversibly inactivated by changes in chromatin structure (chromatin is the complex of DNA and proteins that make chromosomes). In yeast, genes involved in the control of mating type are found in these silent regions, and SIR2 represses their expression by maintaining a silent-competent chromatin structure in this region. The discovery of a Candida albicans SIR2 that is implicated in phenotypic switching suggests that it too has silent regions controlled by SIR2, in which the phenotype-specific genes may perhaps reside.

Another potential regulatory molecule is Efg1p, a transcription factor found in the WO-1 strain that regulates dimorphism, and more recently has been suggested to help regulate phenotypic switching. Efg1p is expressed only in the white and not in the gray cell-type, and overexpression of Efg1p in the gray form causes a rapid conversion to the white form.

So far there are few data that says that dimorphism and phenotypic switching use common molecular components. However, it is not inconceivable that phenotypic switching may occur in response to some change in the environment as well as being a spontaneous event. How SIR2 itself is regulated in S. cerevisiae may yet provide clues as to the switching mechanisms of Candida albicans.

One of the most interesting features of the Candida albicans genome is the occurrence of numeric and structural chromosomal rearrangements as means of generating genetic diversity, named chromosome length polymorphisms (contraction/expansion of repeats), reciprocal translocations, chromosome deletions and trisomy of individual chromosomes. These karyotypic alterations lead to changes in the phenotype, which is an adaptation strategy of this fungus. These mechanisms will be better understood with the complete analysis of the Candida albicans genome.

The Candida albicans genome is being sequenced at the Stanford DNA Sequencing and Technology Center. Funding for this project is provided by National Institute of Dental and Craniofacial Research and Burroughs-Wellcome Fund. A pilot sequencing program is also being carried on by The Sanger Center. Funding for this project is provided by Beowulf Genomics.

Saccharomyces cerevisiae is a species of budding yeast. It is perhaps the most relevant yeast for mankind, both for its use since ancient times in baking and brewing, and for being one of the most intensively studied eukaryotic model organisms in molecular and cell biology.

Saccharomyces cerevisiae was the first eukaryote to have its genome sequenced (published in 1996). The genome is composed of about 13,000,000 base pairs and 6,275 genes. It is estimated that yeast shares about 23% of its genome with humans.

Schizosaccharomyces pombe, also called "fission yeast," is a species of yeast. It is used as a model organism in molecular and cell biology. It is a unicellular eukaryote, whose cells are rod-shaped. These cells maintain their shape by growing exclusively through the cell tips and divide by medial fission to produce two daughter cells of equal sizes.

Fission yeast was isolated in 1893 by Lindner from East African beer. The species name is derived from the Swahili word for beer (Pombe).

The sequence of the S. pombe genome was published in 2002.

The fission yeast researcher Paul Nurse won the 2001 Nobel Prize in Physiology or Medicine, together with Lee Hartwell and Tim Hunt, for work on the cell cycle.

Yeast, any of a number of microscopic, one-celled fungi important for their ability to ferment carbohydrates in various substances. Yeasts in general are widespread in nature, occurring in the soil and on plants. Most cultivated yeasts belong to the genus Saccharomyces; those known as brewer's yeasts are strains of Saccharomyces cerevisiae. For a discussion of the physiology and reproductive behaviour of yeasts, see Fungi: Classification: Ascomycota.

Yeasts have been used since prehistoric times in the making of bread and wine, but their cultivation and use in large quantities was put on a scientific basis by the work of the French microbiologist Louis Pasteur in the 19th century. Today they are used industrially in a wide range of fermentation processes; medicinally, as a source of B-complex vitamins and thiamine and as a stage in the production of various antibiotics and steroid hormones; and as animal feed and foodstuff for humans.

Pure yeast cultures are grown in a medium of sugars, nitrogen sources, minerals, and water. The final product may take the form of dried yeast cells, or the yeast may be pressed into cakes with some starchy material. When a batch of yeast for baking, medicinal, or food purposes is completed, the medium in which the yeast was grown is discarded. In the making of wines, beers, spirits, and industrial alcohol, however, the fermented medium is the desired product, and the yeast itself is discarded or used to make animal feed. See also Brewing.

A baker's yeast is a yeast culture which has been bred specifically for the purpose of leavening breads.

Members of the Division Ascomycota are known as the Sac Fungi and are fungi that produce spores in a distinctive type of microscopic sporangium called an ascus (Greek for a "bag" or "wineskin"). This monophyletic grouping was formerly known as the Ascomycetae and is an extremely significant and successful group of organisms (12,000 species in 1950), accounting for some 75% of all described fungi. Included are most of the fungi that combine with algae to form lichens. The majority of fungi that lack morphological evidence of sexual reproduction are placed here. Better known examples of sac fungi are yeasts, morels, truffles, and Penicillium. The majority of plant-pathogenic fungi belong to this group, or the related Deuteromycota. Species of ascomycetes are also popular in the laboratory. Sordaria fimicola, Neurospora crassa and several species of yeasts are used in many genetics and cell biology experiments.

An ascomycete produces great numbers of asci at any one time, and these may be contained in a structure called an ascocarp. Each ascus contains eight (or a multiple of 8) ascospores, the result of one round of mitosis following meiosis. The resulting haploid nuclei are surrounded by membranes (from the plasma membrane in Euascomycetes; from the nuclear membrane in Hemiascomycetes) and eventually a spore wall.

An exception to the structure described above are the yeasts, which are secondarily unicellular.

An ascospore is a spore contained in an ascus or that was produced inside an ascus. This kind of spore is specific to ascomycetes.

Yeast are small single cell organisms. It is everywhere you can imagine. When making beer the yeast we are referring to is Saccharomyces cerevisiae for ales and Saccharomyces uvarum for lagers. There are many strains of this yeast to obtain different flavor profiles. There are other types of yeast and bacteria that one can use in brewing. Lambic styles call for some different bacteria that wouldn't normally be used in making beer, thus lending the sour taste that lambics are known for. Yeast can be purchased in dry or liquid suspension forms. The liquid yeast gives the brewer more control over the flavor profile. You can also culture your own yeast from starters or obtained from other beers.

Ale yeast, which is top fermenting, tends to make a soft or sometimes fruity beer, while lager yeast, which is bottom fermenting, can make it dry and crisp. Ales ferment at higher temperatures (66 - 74° F) than do lagers (45 - 65° F). The lower temperature prevents the fruity flavors from being absorbed.

You may find some discrepancies in the yeast names. In some places the name Saccharomyces carlsbergensis is used. This is just an older name for Saccharomyces uvarum. In other places, you may not see either the calsbergenesis or uvarum names. Recently the powers that be did some regrouping of the names and put them together under cerevisiae.

As yeast feeds on the sugars in the wort they reproduce. After fermentation starts a layer of foam will be present in the fermenting vessel. This will soon turn into a rocky, dirty looking head that can vary in size. A lot of carbon dioxide is being produced and if the fermenter is not able to expel this it can explode. A blow off tube may be a good idea if you have high gravity wort.

Yeasts are single-celled fungi. As fungi, they are related to the other fungi that people are more familiar with. These include edible mushrooms available at the supermarket, common baker’s yeast used to leaven bread, molds that ripen blue cheese and the molds that produce antibiotics for medical and veterinary use. Many consider edible yeast and fungi to be as natural as fruits and vegetables.

Over 600 different species of yeast are known and they are widely distributed in nature. They are found in association with other microorganisms as part of the normal inhabitants of soil, vegetation, marine and other aqueous environments. Some yeast species are also natural inhabitants of man and animals. While some species are highly specialized and found only in certain habitats at certain times of the year, other species are generalists and can be isolated from many different sources.

Baker’s yeast is used to leaven bread throughout the world and it is the type of yeast that people are most familiar with. Baker’s yeast is produced from the genus and species of yeast called Saccharomyces cerevisiae. The scientific name of the genus of baker’s yeast, Saccharomyces, refers to “saccharo” meaning sugar and “myces” meaning fungus. The species name, cerevisiae, is derived from the name Ceres, the Roman goddess of agriculture. Baker’s yeast products are made from strains of this yeast selected for their special qualities relating to the needs of the baking industry.

The typical yeast cell is approximately equal in size to a human red blood cell and is spherical to ellipsoidal in shape. Because of its small size, it takes about seven billion yeast cells to make up to one gram of compressed baker’s yeast. Yeast reproduce vegetatively by budding, a process during which a new bud grows from the side of the existing cell wall. This bud eventually breaks away from the mother cell to form a separate daughter cell. Each yeast cell, on average, undergoes this budding process 12 to 15 times before it is no longer capable of reproducing. During commercial production, yeast is grown under carefully controlled conditions on a sugar containing media typically composed of beet and cane molasses. Under ideal growth conditions a yeast cell reproduces every two to three hours.

Yeast is the essential ingredient in many bakery products. It is responsible for leavening the dough and imparting a delicious yeast fermentation flavor to the product. It is used in rather small amounts in most bakery products, but having good yeast and using the yeast properly often makes the difference between success and something less than success in a bakery operation.

Candida albicans is a dimorphic fungus that grows at 37oC. Its normal habitat is the mucosal membranes of humans and other warm-blooded animals, where it grows as a yeast (Fig. C) and causes little or no damage. In fact, it can be isolated from the mucosa of up to 50% of humans - from the mouth, the gut, the vagina or, less often, from the surface of the skin.

In some circumstances, however, the same strains of C. albicans that grow as harmless commensals can become pathogenic, invading the mucosa and causing significant damage. This usually happens when a variety of predisposing factors cause the yeast population to multiply, escaping the normal competition from resident bacteria which keep the yeast population in check. Then the yeast cells sprout a hyphal outgrowth (Fig. D) which locally penetrates the mucosal membrane, causing irritation and shedding of the tissues.

One of the best examples of this is the disease termed thrush - a white speckling of the tongue and the back of the throat, resembling the speckling on the bird's chest. This is common in newborn babies, perhaps resulting from passage through an infected birth canal. It is also common in AIDS patients and people who have had a prolonged course of antibacterial therapy, reducing the normal resident bacterial population.

C. albicans also causes vaginitis - inflammation and invasion of the vaginal mucosa, especially during the third trimester of pregnancy and in women who take the pill. The predisposing factors seem to be hormonal, associated with changes in the balance of cell types in the lining epithelium of the vagina. A similar condition termed stomatitis is common in people who wear dentures. Candida can adhere to denture resin, and high sugar levels in the diet can also increase the adhesion by enhancing the production of a mannoprotein adhesive on the yeast cell surface. Systemic candidosis is a more serious condition, when yeast cells proliferate in the circulatory system. This can occur after invasive surgical techniques, including the insertion of intravenous catheters to which the yeast cells adhere, providing a base from which the cells can bud and be disseminated.

All these examples illustrate that C. albicans is a classic opportunistic pathogen, normally kept in check but capable of flaring up in specific, predisposing conditions. It can be identified quite readily from clinical specimens by its ability to sprout hyphae when yeast cells at 37oC are transferred to tubes of horse serum and incubated for 3-5 hours (Fig. D). Only C. albicans and a few related pathogenic Candida species do this. But the fungus has a strong tendency to revert to the yeast phase after only a short period of hyphal growth. Figures E and F show this for horse serum incubated for 24 hours - the hyphae themselves have a beaded appearance, and they give rise to budding yeast cells at the sites where the hyphae of other fungi would form branches.

Cryptococcus albidus is another budding yeast, shown here by phase-contrast microscopy but also with negative staining (the cells are suspended in India ink). Various stages of bud development are seen. The cells are surrounded by a rigid polysaccharide capsule, typical of the genus Cryptococcus, and seen as distinct haloes where the India ink particles have been excluded.

Cryptococcus species are common on leaf surfaces. But the most important species from the human standpoint is C. neoformans, a significant pathogen of immunocompromised people, causing the disease termed cryptococcosis. This disease occurs in about 7-8% of AIDS patients in the USA, and a slightly smaller percentage (3-6%) in western Europe. The capsule is a significant virulence determinant of C. neoformans because it helps to prevent the cells from being recognised and engulfed by white blood cells. Additionally, C. neoformans is unique among Cryptococcus species in producing a phenoloxidase. This enzyme acts on phenolic compounds to produce melanin, which might help to protect the cells against the antimicrobial effects of oxidants in host tissues.

C. neoformans grows commonly on old "weathered" bird droppings in cities, but does not compete well with bacteria in wet droppings. It infects through the lungs, where it causes a mild or chronic, persistent pneumonia, depending on the person's degree of immunity. Random testing of people for skin reactions to C. neoformans antigens in Britain, Australia and the USA indicates that many people have unknowingly been exposed to the fungus with no serious effect. However, in a small proportion of the population the fungus can disseminate "silently" in the central nervous system, causing fatality.

For many years it was assumed that yeast cells inhaled in dried, powdered bird droppings were the source of lung infection. But a sexual stage of the fungus has now been discovered in laboratory culture; it is typical of the fungal group Basidiomycota (which includes the mushroom fungi) but is microscopic, and it leads to the release of small (about 3 micrometre) airborne basidiospores. These are the ideal size for deposition in the lungs (see Airborne Microbes). They are thought to be the main means of infection, but their environmental source is unknown - perhaps a yeast stage growing on vegetation.

Saccharomyces cerevisiae is the budding yeast used for bread-making, where the carbon dioxide produced by growth in the dough causes the bread to rise. Essentially similar yeasts, but now given different species names, are used for production of beers, wines and other alcoholic drinks. This phase-contrast micrograph shows cells in various stages of budding. The buds are small at first, but enlarge progressively and eventually separate from the mother cell by formation of a septum (cross wall).

Few of the cellular organelles can be seen by light microscopy, unless they are stained specifically. The only conspicuous organelle seen in Fig. A is the large central vacuole which contributes to cell expansion. S. cerevisiae is a member of the fungal group Ascomycota (the ascus-forming fungi).

Yeasts are fungi that grow as single cells, producing daughter cells either by budding (the budding yeasts) or by binary fission (the fission yeasts). They differ from most fungi, which grow as thread-like hyphae. But this distinction is not a fundamental one, because some fungi can alternate between a yeast phase and a hyphal phase, depending on environmental conditions. Such fungi are termed dimorphic (with two shapes) and they include several that cause disease of humans.

Yeasts grow typically in moist environments where there is a plentiful supply of simple, soluble nutrients such as sugars and amino acids. For this reason they are common on leaf and fruit surfaces, on roots and in various types of food. With few exceptions, they are unable to degrade polymers, such as starch and cellulose which are used by many hyphal fungi.

Yeast are unicellular fungi. The precise classification is a field that uses the characteristics of the cell, ascospore and colony. Physiological characteristics are also used to identify species. One of the more well known characteristics is the ability to ferment sugars for the production of ethanol. Budding yeasts are true fungi of the phylum Ascomycetes, class Hemiascomycetes. The true yeasts are separated into one main order Saccharomycetales.

Yeasts are characterized by a wide dispersion of natural habitats. Common on plant leaves and flowers, soil and salt water. Yeasts are also found on the skin surfaces and in the intestinal tracts of warm-blooded animals, where they may live symbiotically or as parasites. The common "yeast infection" is typically Candidiasis is caused by the yeast-like fungus Candida albicans. In addition to being the causative agent in vaginal yeast infections Candida is also a cause of diaper rash and thrush of the mouth and throat.

Yeasts multiply as single cells that divide by budding (eg Saccharomyces) or direct division (fission, eg. Schizosaccharomyces), or they may grow as simple irregular filaments (mycelium). In sexual reproduction most yeasts form asci, which contain up to eight haploid ascospores. These ascospores may fuse with adjoining nuclei and multiply through vegetative division or, as with certain yeasts, fuse with other ascospores.

The awsome power of yeast genetics is partially due to the ability to quickly map a phenotype producing gene to a region of the S. cerevisiae genome. For the past two decades S. cerevisiae has been the model system for much of molecular genetic research because the basic cellular mechanics of replication, recombination, cell division and metabolism are generally conserved between yeast and larger eukaryotes, including mammals.

The most well-known and commercially significant yeasts are the related species and strains of Saccharomyces cerevisiae. These organisms have long been utilized to ferment the sugars of rice, wheat, barley, and corn to produce alcoholic beverages and in the baking industry to expand, or raise, dough. Saccharomyces cerevisiae is commonly used as baker's yeast and for some types of fermentation. Yeast is often taken as a vitamin supplement because it is 50 percent protein and is a rich source of B vitamins, niacin, and folic acid.

In brewing, Saccharomyces carlsbergensis, named after the Carlsberg Brewery in Copenhagen, where it was first isolated in pure culture by Dr. Emil Christian Hansen (1842-1909) in 1883, is used in the production of several types of beers including lagers. S. carlsbergensis is used for bottom fermentation. S. cerevisiae used for the production of ales and conducts top fermentation, in which the yeast rise to the surface of the brewing vessel. In modern brewing many of the original top fermentation strains have been modified to be bottom fermenters. Currently the S. carlsbergensis designation is not used, the S. cerevisiae classification is used instead.

The yeast's function in baking is to ferment sugars present in the flour or added to the dough. This fermentation gives off carbon dioxide and ethanol. The carbon dioxide is trapped within tiny bubbles and results in the dough expanding, or rising. Sourdough bread, is not produced with baker's yeast, rather a combination of wild yeast (often Candida milleri) and an acid-generating bacteria (Lactobacillus sanfrancisco sp. nov). It has been reported that the ratio of wild yeast to bacteria in San Francisco sourdough cultures is about 1:100. The C. milleri strengthens the gluten and the L. sanfrancisco ferments the maltose. For more information about sourdough see rec.food.sourdough FAQ.

The fermentation of wine is initiated by naturally occurring yeasts present in the vineyards. Many wineries still use nature strains, however many use modern methods of strain maintenance and isolation. The bubbles in sparkling wines is trapped carbon dioxide, the result of yeast fermenting sugars in the grape juice. One yeast cell can ferment approximately its own weight of glucose per hour. Under optimal conditions S. cerevisiae can produce up to 18 percent, by volume, ethanol with 15 to 16 percent being the norm. The sulfur dioxide present in commercially produced wine is actually added just after the grapes are crushed to kill the naturally present bacteria, molds, and yeasts.

The yeastlike fungus, Candida albicans, is commonly found in the mouth, vagina, and intestinal tract. Candida is a normal inhabitant of humans and normally causes no ill effects. However, among infants and individuals with other illness a variety of conditions can occur. Candidiasis of the mucous membranes of the mouth is known as thrush. Candidiasis of the vagina is called vaginitis. Candida also causes severe disease in persons with AIDS and chemotherapy patients.

Budding yeasts are true fungi of the phylum Ascomycetes, class Hemiascomycetes. The true yeasts are separated one main order Saccharomycetales, which includes at least ten families. Yeasts are heterotrophic, lack chlorophyll, and are characterized by a wide dispersion of natural habitats. Common on plant leaves and flowers, yeasts are also found on the skin surfaces and in the intestinal tracts of warm-blooded animals, where they may live symbiotically or as parasites. k, g, d, b, l. In women, who are pregnant or taking antibiotics, an infection of the vagina and vulva caused by a yeastlike fungus Candida albicans, is common. Yeasts are also found in soil and saltwater, where they contribute to the decomposition of plant and algal matter.

Are yeasts Fungi?

Yes, the 700 known yeasts really are fungi, although their colonies often look more like those of bacteria.

Most fungi explore their surroundings by producing miles of fine, branching filaments called hyphae, but most yeasts have become more or less unicellular, with rounded cells. This is often an adaptation to living in a liquid medium of high osmotic pressure. This usually means media with a high sugar content, such as is found in the nectaries of flowers or on the surface of fruits, where if they present the least possible surface area (as close to spherical as possible), it makes it easier for them to control the movement of dissolved substances in and out of their cells.

The cells of most yeasts can be regarded as asexual propagules, and they produce more of the same by a variety of methods similar to those found in moulds.

Some yeasts are related to ascomycetes, others to basidiomycetes, and even zygomycetes sometimes take on a yeast-like appearance.

Some yeasts make hyphae as well as unicells, and some are even exclusively hyphal, being recognizable as yeasts only by biochemical characters.

Yeasts are, of course, among the most important fungi, because they raise bread, ferment sugars to make beer, wine, and spirits, and represent a concentrated food and a source of B vitamins.

A few yeasts cause serious diseases of people.

Yeast name applied specifically to a certain group of microscopic fungi and to commercial products consisting of masses of dried yeast cells or of yeast mixed with a starchy material and pressed into yeast cakes. Although a number of fungi are sometimes called yeasts, the true yeasts are unicellular, consist of oval or round cells, and reproduce chiefly by budding. Under certain conditions some yeast cells secrete a thickened wall, and the cytoplasm of the single cell within divides to form four or eight cells, or spores, known as ascospores, which emerge when the wall ruptures. In a few species two cells fuse before undergoing spore formation. There are about 500 species in all. 1 Yeasts, especially those of the genus Saccharomyces, have long been of commercial importance because they are the chief agents in alcoholic fermentation. Because of this they are essential to the making of beer, wine, and other alcoholic beverages and industrial alcohol. Wild yeasts, those found in nature and probably carried by insects from the soil to fruits, are frequently active in the fermentation process. In breadmaking the yeasts act upon the carbohydrates in the dough, forming carbon dioxide and ethyl alcohol, which are driven off in the baking process; the escaping carbon dioxide causes the bread to rise. Since early times yeast has been used in treating various ailments; brewer’s yeast has a high content of thiamine and other vitamins of the B-complex group. Yeasts are classified in the kingdom Fungi, phyla (divisions) Ascomycota and Basidiomycota.

Yeast is a living, microscopic, single-cell organism that, as it grows, converts its food (through a process known as fermentation) into alcohol and carbon dioxide. This trait is what endears yeast to winemakers, brewmasters and breadbakers. In the making of wine and beer, the yeast's manufacture of alcohol is desired and necessary for the final product; and carbon dioxide is what makes BEER and CHAMPAGNE effervescent. The art of breadmaking needs the carbon dioxide produced by yeast in order for certain doughs to rise. To multiply and grow, all yeast needs is the right environment, which includes moisture, food (in the form of sugar or starch) and a warm, nurturing temperature (70° to 85°F is best). Wild yeast spores are constantly floating in the air and landing on uncovered foods and liquids. No one's sure when these wild spores first interacted with foods but it's known that the Egyptians used yeast as a LEAVENING agent more than 5,000 years ago. Wine and other fermented beverages were made for millennia before that. Today, scientists have been able to isolate and identify the various yeasts that are best for winemaking, beermaking and baking. The two types commercially available are baker's yeast and brewer's yeast. Baker's yeast, as the name implies, is used as a leavener. It's catagorized into three basic types-active dry yeast, compressed fresh yeast and YEAST STARTERS. Active dry yeast is in the form of tiny, dehydrated granules. The yeast cells are alive but dormant because of the lack of moisture. When mixed with a warmliquid (105° to 115°F), the cells once again become active. Active dry yeast is available in two forms, regular and quick-rising, of which the latter takes about half as long to leaven bread. They may be used interchangeably (with adjustments in rising time) and both are available in 1/4-ounce envelopes. Regular active dry yeast may also be purchased in 4-ounce jars or in bulk in some health-food stores. It should be stored in a cool, dry place, but can also be refrigerated or frozen. It should always be at room temperature before being dissolved in liquid. Properly stored, it's reliable when used by the expiration date, which should be stamped on the envelope or jar label. One package of dry yeast is equal to 1 scant tablespoon dry yeast or 1 cake of compressed fresh yeast. Compressed fresh yeast, which comes in tiny (0.06-ounce), square cakes, is moist and extremely perishable. It must be refrigerated and used within a week or two, or by the date indicated on the package. It can be frozen, but should be defrosted at room temperature and used immediately. One cake of fresh yeast can be substituted for one envelope of dry yeast. The use of compressed fresh yeast has been primarily replaced by the more convenient active dry yeast. All baker's yeast should be given a test called proofing to make sure it's still alive. To proof yeast, dissolve it in warm water and add a pinch of sugar. Set the mixture aside in a warm place for 5 to 10 minutes. If it begins to swell and foam, the yeast is alive, active and capable of leavening bread. Brewer's yeasts are special non-leavening yeasts used in beermaking. Because it's a rich source of B vitamins, brewer's yeast is also used as a food supplement. It's available in health-food stores. Brewer's yeasts are also marketed in specialty beermaking equipment shops, with different strains used for different beers.

Prior to the evolution of commercially available baking powders and yeasts during the 19th century, yeast starters were the LEAVENERS used in breadmaking. Such starters are a simple mixture of flour, water, sugar and YEAST. (At one time, airborne yeast was the only source used, but today convenient commercially packaged baker's yeast is more common.) This batter is set aside in a warm place until the yeast ferments and the mixture is foamy. A portion of the starter-usually about 2 cups-is removed and used as the base and leavener for some bread recipes. Once fermented, yeast starters-the most famous of which is sourdough starter-can be kept going in the right environment for years simply by adding equal parts flour and water. Herman starter is a colloquialism (of unknown origin) for a honey- or sugar-sweetened starter used primarily for sweet breads. Starter should be refrigerated and can be stored this way indefinitely as long as it's replenished every 2 weeks. Before using or replenishing, it should be brought to room temperature. If a starter turns orange or pink and develops an unpleasantly acrid odor, undesirable bacteria have invaded it and the mixture must be discarded. Two cups of the foamy starter mixture can be substituted for each package of yeast called for in a recipe.

yeast, name applied specifically to a certain group of microscopic fungi and to commercial products consisting of masses of dried yeast cells or of yeast mixed with a starchy material and pressed into yeast cakes. Although a number of fungi are sometimes called yeasts, the true yeasts are unicellular, consist of oval or round cells, and reproduce chiefly by budding. Under certain conditions some yeast cells secrete a thickened wall, and the cytoplasm of the single cell within divides to form four or eight cells, or spores, known as ascospores, which emerge when the wall ruptures. In a few species two cells fuse before undergoing spore formation. There are about 500 species in all.

Yeasts multiply as single cells that divide by budding or direct division (fission), or they may grow as simple irregular filaments (mycelium). In sexual reproduction most yeasts form asci, which contain up to eight haploid ascospores. These ascospores may fuse with adjoining nuclei and multiply through vegetative division or, as with certain yeasts, fuse with other ascospores. The awsome power of yeast genetics is partially due to the ability to quickly map a phenotype producing gene to a region of the S. cerevisiae genome. For the past two decades S. cerevisiae has been the model system for much of molecular genetic research because the basic cellular mechanics of replication, recombination, cell division and metabolism are generally conserved between yeast and larger eukaryotes, including mammals. The most well-known and commercially significant yeasts are the related species and strains of Saccharomyces cerevisiae. k, b, b, c, k. These organisms have long been utilized to ferment the sugars of rice, wheat, barley, and corn to produce alcoholic beverages and in the baking industry to expand, or raise, dough. Saccharomyces cerevisiae is commonly used as baker's yeast and for some types of fermentation. Yeast is often taken as a vitamin supplement because it is 50 percent protein and is a rich source of B vitamins, niacin, and folic acid.

Yeasts, especially those of the genus Saccharomyces, have long been of commercial importance because they are the chief agents in alcoholic fermentation. Because of this they are essential to the making of beer, wine, and other alcoholic beverages and industrial alcohol. Wild yeasts, those found in nature and probably carried by insects from the soil to fruits, are frequently active in the fermentation process. In breadmaking the yeasts act upon the carbohydrates in the dough, forming carbon dioxide and ethyl alcohol, which are driven off in the baking process; the escaping carbon dioxide causes the bread to rise. Since early times yeast has been used in treating various ailments; brewer's yeast has a high content of thiamine and other vitamins of the B-complex group. Yeasts are classified in the kingdom Fungi, phyla (divisions) Ascomycota and Basidiomycota.

Yeast is a living, microscopic, single-cell organism that, as it grows, converts its food (through a process known as fermentation) into alcohol and carbon dioxide. This trait is what endears yeast to winemakers, brewmasters and breadbakers. In the making of wine and beer, the yeast's manufacture of alcohol is desired and necessary for the final product; and carbon dioxide is what makes beer and champagne effervescent. The art of breadmaking needs the carbon dioxide produced by yeast in order for certain doughs to rise. To multiply and grow, all yeast needs is the right environment, which includes moisture, food (in the form of sugar or starch) and a warm, nurturing temperature (70 degrees to 85 degrees F is best). Wild yeast spores are constantly floating in the air and landing on uncovered foods and liquids. No one's sure when these wild spores first interacted with foods but it's known that the Egyptians used yeast as a leavening agent more than 5,000 years ago. Wine and other fermented beverages were made for millennia before that. Today, scientists have been able to isolate and identify the various yeasts that are best for winemaking, beermaking and baking. The two types commercially available are baker's yeast and brewer's yeast. Baker's yeast, as the name implies, is used as a leavener. It's catagorized into three basic types--active dry yeast, compressed fresh yeast and yeast starters. Active dry yeast is in the form of tiny, dehydrated granules. The yeast cells are alive but dormant because of the lack of moisture. When mixed with a warm liquid (105 degrees to 115 degrees), the cells once again become active. Active dry yeast is available in two forms, regular and quick-rising, of which the latter takes about half as long to leaven bread. They may be used interchangeably (with adjustments in rising time) and both are available in 1/4-ounce envelopes. Regular active dry yeast may also be purchased in 4-ounce jars or in bulk in some health-food stores. It should be stored in a cool, dry place, but can also be refrigerated or frozen. It should always be at room temperature before being dissolved in liquid. Properly stored, it's reliable when used by the expiration date, which should be stamped on the envelope or jar label. One package of dry yeast is equal to 1 scant tablespoon dry yeast or 1 cake of compressed fresh yeast. Compressed fresh yeast, which comes in tiny (0.6-ounce), square cakes, is moist and extremely perishable. It must be refrigerated and used within a week or two, or by the date indicated on the package. It can be frozen, but should be defrosted at room temperature and used immediately. One cake of fresh yeast can be substituted for one envelope of dry yeast. The use of compressed fresh yeast has been primarily replaced by the more convenient active dry yeast. All baker's yeast should be given a test called proofing to make sure it's still alive. To proof yeast, dissolve it in warm water and add a pinch of sugar. Set the mixture aside in a warm place for 5 to 10 minutes. If it begins to swell and foam, the yeast is alive, active and capable of leavening bread. Brewer's yeasts are special non-leavening yeasts used in beermaking. Because it's a rich source of B vitamins, brewer's yeast is also used as a food supplement. It's available in health-food stores. Brewer's yeasts are also marketed in specialty beermaking equipment shops, with different strains used for different beers.

Budding yeasts are true fungi of the phylum Ascomycetes, class Hemiascomycetes. The true yeasts comprise the family Saccharomycetes, which has but one genus Saccharomyces, but includes at least ten species. The classification of yeasts is a specialized field using cell, ascospore, and colony characteristics for distinguishing genera, and physiological characteristics - particularly the ability to ferment individual sugars - to identify species. Yeasts are heterotrophic, lack chlorophyll, and are characterized by a wide dispersion of natural habitats. Common on plant leaves and flowers, yeasts are also found on the skin surfaces and in the intestinal tracts of warm-blooded animals, where they may live symbiotically or as parasites. In women, who are pregnant or taking antibiotics, an infection of the vagina and vulva caused by a yeast like fungus Candida albicans, is common. Yeasts are also found in soil and saltwater, where they contribute to the decomposition of plant and algal matter. Yeasts multiply as single cells that divide by budding or direct division, or they may grow as simple irregular filaments. In sexual reproduction most yeasts form asci, which contain up to eight haploid ascospores. These ascospores may fuse with adjoining nuclei and multiply through vegetative division or, as with certain yeasts, fuse with other ascospores. The most well-known and commercially significant yeasts are the related species and strains of Saccharomyces cerevisiae. These organisms have long been utilized to ferment the sugars of rice, wheat, barley, and corn to produce alcoholic beverages and in the baking industry to exp