J Bacteriol, 1979 May, 138(2), 370 - 6 Characterization of a succinate dehydrogenase complex solubilized from the cytoplasmic membrane of Bacillus subtilis with the nonionic detergent Triton X-100; Hederstedt L et al.; A succinic dehydrogenase (SDH) complex has been purified from Triton X-100-solubilized membranes from Bacillus subtilis by precipitation with specific antibody . Radioactively labeled precipitated complex was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography of the gels . The complex contained equimolar amounts of three polypeptides with approximate molecular weights of 65,000, 28,000, and 19,000 . Five succinic dehydrogenase-negative mutants, belonging to the citF group, contained the 65,000-dalton polypeptide in a soluble form in the cytoplasm . Each 65,000-dalton polypeptide had about one molecule of flavin bound . Another citF mutant, citF11, which lacks the 65,000-dalton polypeptide, contained a membrane-bound 28,000-dalton polypeptide . The wild-type succinic dehydrogenase complex contained cytochrome, probably a cytochrome b . The 19,000-dalton polypeptide is suggested to represent the apoprotein of this cytochrome . The 65,000-dalton and the 28,000-dalton polypeptides are thought to constitute succinic dehydrogenase and to correspond to the flavoprotein and the ironprotein, respectively, as described for succinic dehydrogenase isolated from beef heart mitochondria or Rhodospirillum rubrum chromatophores . The results presented suggest that in B . subtilis succinic dehydrogenase is attached to a cytochrome b in the membrane via the 28,000-dalton (ironprotein) polypeptide. J Bacteriol, 1979 May, 138(2), 345 - 51 Export of extracellular levansucrase by Bacillus subtilis: inhibition by cerulenin and quinacrine; Caulfield MP et al.; Bacillus subtilis B secretes an inducible, extracellular enzyme, levansucrase . Inhibition studies were undertaken to investigate the possible mechanism of release of this enzyme . The antibiotic cerulenin, at a concentration of 10 micrograms/ml, totally inhibited de novo lipid synthesis in B . subtilis B for at least 1 h, while only slightly reducing protein and RNA synthesis . At this concentration cerulenin, added concomitantly with the inducer sucrose, prevented the release of levansucrase for at least 150 min . This was not due to the prevention of inducer uptake by the cells . The release of the enzyme was also independent of cell division . In B . subtilis 1007 the induction of beta-galactosidase by 5 mM lactose was not prevented by cerulenin . Preliminary evidence indicated the association of a lipid moiety with the enzyme as it passes through the cytoplasmic membrane . Quinacrine (0.2 mM), which inhibits the penicillinase-releasing protease of Bacillus licheniformis, inhibited levansucrase release from B . subtilis B, but had no effect on lipid synthesis. J Bacteriol, 1979 May, 138(2), 314 - 9 Specific inhibition of outgrowth of Bacillus subtilis spores by novobiocin; Gottfried M et al.; Spores of a Bacillus subtilis mutant temperature sensitive in deoxyribonucleic acid (DNA) replication proceeded through outgrowth at the nonpermissive temperature to the same extent as the wild-type parent spores . In contrast, the DNA synthesis inhibitor novobiocin completely prevented spore outgrowth while displaying a marginal effect on logarithmic growth during one generation time . Inhibition of outgrowth by novobiocin occurred in the absence of DNA replication, as demonstrated in an experiment with spores of the temperature-sensitive DNA synthesis mutant at the restrictive temperature . Novobiocin inhibited the initial rate of ribonucleic acid synthesis to the same extent in germinated spores and in exponentially growing cells . A novobiocin-resistant mutant underwent normal outgrowth in the presence of novobiocin . Therefore, novobiocin inhibition was independent of its effect on chromosome replication per se. Biochim Biophys Acta, 1979 Apr 19, 552(3), 558 - 62 Interactions between bacterial membranes and peptidolipids: lysis of micrococcus luteus protoplasts by derivatives of peptidolipidic antibiotics from bacillus subtilis; Besson F et al.; The lysis of protoplasts of Micrococcus luteus has been tested with various derivatives of three peptidolipidic antibiotics: iturin A, mycosubtilin and bacillomycin L . The lytic activity is dependent to the nature of the substituting group and to the position of the substituted aminoacid residue . The acetylation of OH groups leads to a decrease of the lytic activity of the natural antibiotics . The methylation of aspartyl residues of bacillomycin L gives a strong lytic activity while natural bacillomycin L has no lytic activity . The methylation of the tyrosyl residue enhances the lytic activities of iturin A and of bacillomycin L-dimethyl ester and reduces that of mycosubtilin . Correlations between the structures of derivatives and their lytic action on M . luteus protoplasts are discussed. Biochim Biophys Acta, 1979 Apr 19, 552(3), 492 - 8 Characterization of glutamate transport system in hydrophobic protein (H protein) of Bacillus subtilis; Kusaka I et al.; Hydrophobic protein (H protein) was isolated from membrane fractions of Bacillus subtilis and constituted into artificial membrane vesicles with lipid of B . substilis . Glutamate was accumulated into the vesicle when a Na+ gradient across the membrane was imposed . The maximum effect of Na+ on the transport was achieved at a concentration of about 40 mM, while the apparent Km for Na+ was approximately 8 mM . On the other hand, Km for glutamate in the presence of 50 mM Na+ was about 8 micro M . Increasing the concentration of Na+ resulted in a decrease in Km for glutamate, maximum velocity was not affected . The transport was sensitive to monensin (Na+ ionophore) . Glutamate was also accumulated when pH gradient (interior alkaline) across the membrane was imposed or a membrane potential was induced with K+-diffusion potential . The pH gradient-driven glutamate transport was sensitive to carbonylcyanide m-chlorophenylhydrazone and the apparent Km for glutamate was approximately 25 microM . These results indicate that two kinds of glutamate transport system were present in H protein: one is Na+ dependent and the other is H+ dependent. Experientia, 1979 Apr 15, 35(4), 470 - 2 Different effects of D-glucose anomers for respiration of bacterial germinated spores; Ishihara H et al.; Effects of alpha- or beta-D-glucose on the respiration of germinated spores (only germinated spores not including swollen spores and elongated spores) of Bacillus subtilis and B . megaterium were studied . In our conditions, net amount of oxygen consumed by 10(10) germinated spores of B . subtilis per min after addition of alpha- or beta-D-glucose was 1.6 microgram or 6.6 microgram (beta/alpha = 4.13), while that by B.megaterium was 4.5 microgram of 6.8 microgram (beta/alpha = 1.51), respectively . However, the net amounts of oxygen consumed by 10(10) vegetative cells per min after addition of alpha- or beta-D-glucose were identical, for B.subtillis in both cases 443.0 microgram and for B.megaterium in both cases 604.4 microgram. J Biol Chem, 1979 Apr 10, 254(7), 2184 - 6 Regulation of teichoic acid synthesis during phosphate limitation; Glaser L et al.; Bacillus subtilis W-23, when placed in phosphate-free medium, ceases to synthesize teichoic acid and synthesizes teichuronic acid . The enzymatic basis for the cessation of teichoic acid synthesis is the irreversible inhibition of the first membrane-bound enzyme involved in teichoic acid synthesis which catalyzes the reaction Undecapenol-P + UDP-GlcNAc leads to undecaprenol-P-P-GlcNAc + UMP. J Gen Microbiol, 1979 Apr, 111(2), 353 - 61 Intrastrand self-complementary sequences in Bacillus subtilis DNA; Galloway DA et al.; Intrastrand self-complementary sequences have been isolated from the DNA of Bacillus subtilis by hydroxyapatite (HA) chromatography following thermal renaturation of strands separated by chromatography on methylated albumin kieselguhr (MAK) . The instrastrand structures derived from the MAK H strand (HA HII) were biologically active showing transforming activity for a wide variety of markers, as well as hybridization to both pulse-labelled and ribosomal RNA . Removal of regions of single-strand DNA with S1 nuclease did not significantly alter the biological activity of the self-annealed molecules . The overall efficiency of transformation and hybridization of the intrastrand self-annealing DNA was low suggesting that many sequences in the population are neither active in transformation to prototrophy nor transcribed into RNA. Mol Gen Genet, 1979 Mar 5, 170(3), 345 - 9 Lethal effect of protamine and histone on competent Bacillus subtilis cells . Inhibition of genetic transformation by protamine in sublethal concentration; Antohi S et al.; Under experimental conditions of genetic transformation, protamine and total histone were bactericidal for Bacillus subtilis cells . The abilities to cause lethality were very similar for both, either protamine or histone, with no antagonistic effects amongst these natural polycations . With both basic proteins acting simultaneously the enhancement was higher than a summation of the separate lethal effects . Sublethal concentration of protamine added at the beginning of transformation time, produced a strong inhibition of transforming efficiency . The same concentration added later than 10 min from the start of transformation had no inhibitory effect . These facts together with the absence of inhibition by simple pretreatment of DNA alone as well as the cell protection by protamine against lytic activity of lysozyme, suggest a protamine-cell surface interaction which impedes DNA uptake events. J Bacteriol, 1979 Mar, 137(3), 1452 - 5 Spectinomycin dependence in Bacillus subtilis; Henkin TM et al.; Spectinomycin dependence in Bacillus subtilis involves two mutations, one conferring drug resistance and the other producing a requirement for spectinomycin for growth. J Gen Microbiol, 1979 Mar, 111(1), 165 - 80 Genetics analysis of spore germination mutants of Bacillus subtilis 168: the correlation of phenotype with map location; Moir A et al.; The isolation and characterization of 29 new germination (Ger) mutants of Bacillus subtilis 168 is described . These were classified, along with previously described mutants, into seven groups according to map location . The mutations in 26 GerA mutants mapped between cysB and thr; detailed mapping of two of these has located them very close to citG . These mutants were deficient in germination in alanine, but responded to the germinative combination of asparagine, glucose, fructose and KCl . One GerB mutant mapped on the origin-proximal side of hisA; it was normal in germination in alanine, but deficient in termination in a mixture of asparagine, glucose, fructose and KCl . Two GerC mutants were linked to lys, but were separable from a temperature-sensitive growth deficiency mapping between lys and trp . The GerC mutants had a similar germination phenotype to the GerA mutants . Three GerD mutants did not germinate in either of the above germinants or in Penassay Broth . They were located on the side of ery distal to cysA . The GerE mutant, which did not germinate in any of the three germinants, was located very close to citF and possessed an altered spore coat . The two GerF mutants were defective in germination in all three germinants and mapped on the origin proximal-side of hisA, but much closer to his than did the GerB mutant . A phosphoglycerate kinase-negative mutant altered in germination mapped between cysB and hisA (GerG) . These mutants have established a minimum of seven locations important to germination, and will be useful in the development and appraisal of theories of spore germination. Nucleic Acids Res, 1979 Mar, 6(3), 1203 - 19 Antibody to B . subtilis DNA polymerase III: use in enzyme purification and examination of homology among replication-specific DNA polymerases; Barnes MH et al.; Bacillus subtilis DNA polymerase III (pol III), an arylhydrazinopyrimidine-sensitive, replication-specific enzyme, was used to generate a non-precipitating rabbit antibody which specifically inhibited pol III activity in vitro . The antibody was used to examine structural relationships among several DNA polymerases, and it was linked covalently to agarose; the antibody:agarose was employed to develop a rapid, selective method of purification of catalytically active B . subtilis pol III. Mikrobiologiia, 1979 Mar-Apr, 48(2), 249 - 55 {Effect of exogenous nucleodepolymerases on Bacillus subtilis multiplication}; Kupriianova FG et al.; The effect of exogenous nucleodepolymerases on cell growth was studied with Bacillus subtilis . Nucleodepolymerases of microbial and animal origin stimulated growth of the culture . The stimulating action of DNAases depended on the dose of the enzyme added to the growth medium and on the phase of the cultural growth . Acceleration of the cultural growth correlated with intensification of DNA synthesis in the bacterial cells. J Bacteriol, 1979 Mar, 137(3), 1354 - 61 Electron microscopic study of Bacillus subtilis protoplast fusion; Frehel C et al.; When protoplasts derived from sporulating cells of Bacillus subtilis were fused by exposure to polyethylene glycol (PEG) and fixed immediately thereafter, protoplasts with two enclosed prespores could be seen by electron microscope . The number of fusion events was greatly increased, and multiply fused protoplasts appeared, when the PEG-treated suspension was diluted in hypertonic broth and reincubated before fixation . This post-PEG incubation effect is taken to indicate a fusion mechanism of two steps: a short, PEG-dependent step of membrane activation, followed by a slow, metabolism-requiring step completing fusion . When prespore-bearing protoplasts from two genetically different strains were mixed and fused, the extent of fusion could also be followed by counting clones of recombinant bacteria . Maximal from the start, their number (1% of each parent type protoplast present) was unaffected by post-PEG incubation . Fusion in this case is apparently completed after plating on the wall-regeneration medium . After optimal post-PEG incubation, the majority of the protoplasts were seen to participate in fusion, and the cytological fusion observed, corrected for wall-regeneration frequency, accounted quantitatively for the prototrophic bacteria eventually recovered . These results are in good agreement with those obtained independently by Sanchez-Rivas and Garro (J . Bacteriol . 137:1340--1345, 1979). J Bacteriol, 1979 Mar, 137(3), 1346 - 53 Parameters governing bacterial regeneration and genetic recombination after fusion of Bacillus subtilis protoplasts; Gabor MH et al.; Bacterial protoplast fusion, induced by polyethylene glycol, has been made more regular and convenient by further specification and improvement of various steps in the previously used procedure . These have made it possible to obtain regularly 100% regeneration of Bacillus subtilis cells from protoplasts before treatment with polyethylene glycol and yields of 10 to 75% from polyethylene glycol-treated protoplasts . Genetic recombination frequencies do not increase correspondingly . Also, when regeneration is reduced by various experimental conditions, recombination does not decrease in proportion . It is concluded that regeneration of recombinant-forming cells is independently determined and not closely related to the average regeneration for the population . Kinetic studies with varying individual parental or total protoplast concentrations strongly indicate that protoplast collision and contact is not the limiting factor determining the number of genetic recombinants obtained . Recombination approximates a linear, rather than quadratic, function of the total or of the majority protoplast population present, from which it is concluded that fusion events are always adequate to produce substantially more potential recombinants than are registered . The strong effect of the majority/minority ratio upon the number of minority cells that become recombinant is independent of which parent is in excess . This shows in a direct and physiological way that both parents are equivalent partners in their genetic contributions. J Bacteriol, 1979 Mar, 137(3), 1340 - 5 Bacterial fusion assayed by a prophage complementation test; Sanchez-Rivas C et al.; In previous studies of bacterial protoplast fusion, only the frequencies of cell wall regeneration and of bacterial recombination were determined . In this work the frequency of the heterozygous fusion products is measured by prophage complementation . Two multiply marked nonsuppressing strains of Bacillus subtilis, each lysogenic for a different Sus mutant of the phage phi 105, were induced by mitomycin C, protoplasted, fused, and, after dilution in hypertonic broth, incubated until plating with phi 105-sensitive indicator bacteria . When cell lysis was avoided, the frequency of the heterozygous fused cells could be determined from the number of infectious centers produced . The very high frequencies observed are in good agreement with those determined directly, with nonlysogenic strains, by electron microscopic examination of the fused protoplasts (C . Frehel, A . M . Lheritier, C . Sanchez-Rivas, and P . Schaeffer, J . Bacteriol . 137:1354--1361, 1979) . Evidence is presented that fusion occurs in two steps, one polyethylene glycol dependent, the other energy requiring . The bacterial growth medium affects the ability of the protoplasts to fuse and to regenerate a cell wall . When experiments using different growth media were compared, an inverse relationship between these abilities was observed, and a direct relationship appeared between the heterozygotes (corrected for wall regeneration) and the recombinant bacteria that were found. J Bacteriol, 1979 Mar, 137(3), 1208 - 18 Properties of the Bacillus subtilis spore coat; Pandey NK et al.; About 70% of the protein in isolated Bacillus subtilis spore coats was solubilized by treatment with a combination of reducing and denaturing agents at alkaline pH . The residue, consisting primarily of protein, was insoluble in a variety of reagents . The soluble proteins were resolved into at least seven bands by sodium dodecyl sulfate gel electrophoresis . About one-half of the total was four proteins of 8,000 to 12,000 daltons . These were relatively tyrosine rich, and one was a glycoprotein . There was also a cluster of proteins of about 40,000 daltons and two or three in the 20,000- to 25,000-dalton range . The insoluble fraction had an amino acid composition and N-terminal pattern of amino acids very similar to those of the soluble coat proteins . A major difference was the presence of considerable dityrosine in performic acid-oxidized preparations of insoluble coats . Coat antigen including a 60,000-dalton protein not present in extracts of mature spores was detected in extracts of sporulating cells by immunoprecipitation . This large antigen turned over in a pulse-chase experiment . Antibodies to either the array of 8,000- to 12,000-dalton coat polypeptides or to the larger coat proteins reacted with this 60,000-dalton species, suggesting a common precursor for many of the mature coat polypeptides . Spore coats seem to be assembled by processing of proteins and by secondary modifications including perhaps dityrosine formation for cross-linking. Prikl Biokhim Mikrobiol, 1979 Mar-Apr, 15(2), 314 - 7 {Pigments produced by Bacillus subtilis var . niger 16k}; Sorokulova IB et al.; Pigments produced by Bacillus subtilis var . niger 16k on tyrosine agar and histidine containing synthetic medium were isolated and identified . On the basis of a number of tests the pigments were classified as melanins. Prikl Biokhim Mikrobiol, 1979 Mar-Apr, 15(2), 262 - 8 {Immobilization of Bacillus subtilis proteases on a styrol copolymer with maleic anhydride with covalent and coordination bonds}; Slobodianikova LS et al.; Insoluble enzymes have been obtained through an interaction of the complex preparation of proteases, protosubtilin G15x, with the sterol and maleic anhydride copolymer in aprotic dipolar solvents and as a result of formation of mixed enzyme-polymer complexes of protosubtilin G15x and the sterol and malic acid copolymer in aqueous solution in the presence of metal ions . The effect of metal ions on the yield and activity of the resultant immobilized preparation has been studied. Mol Gen Genet, 1979 Feb 26, 170(2), 123 - 7 Host-controlled modification and restriction in Bacillus subtilis: Bsu 168-system and BsuR-system in B . subtilis 168; Ikawa S et al.; A Bsu168-specific restriction deficient (r168-) mutant of Bacillus subtilis Marburg 168 was transformed to be BsuR-specific restriction proficient (rR+) with B . subtilis R DNA as efficiently as the Bsu 168-specific restriction proficient (r168+) parental strain (hsrM+, hsdR-) . We constructed rR+ mR+ r168+ m168+ strain (ISMR 4), rR+ mR+ r168- m168+ strain (ISR 11) and rR+ mR+ r168- m168- strain (ISR 6) from strain 101 (r168+ m168+), strain 1012 (r168- m168+) and strain RM125 (r168- m168-), respectively by transformation with B . subtilis R DNA, and tested their restriction and modification activities on phage phi 105C . The results show that the sites recognized by Bsu168-specific restriction and modification enzymes and the sites recognized by BsuR-specific ones are not overlapping . We conclude that the Bsu168-modification and restriction system and the BsuR-modification and restriction system are controlled independently by two distinct sets of genes in the rR+ mR+ transformant of r168+ m168+ strain B . subtilis 168. Mol Gen Genet, 1979 Feb 26, 170(2), 117 - 22 Mapping of genes determining nonpermissiveness and host-specific restriction to bacteriophages in Bacillus subtilis Marburg; Saito H et al.; Bacillus subtilis Marburg is nonpermissive for the multiplication of bacteriophages SP10 and phi NR2 . A permissive mutant was derived from the Marburg strain, and the genetic determinants of non-permissiveness were analyzed by PBS1 transduction . The simultaneous presence of two genes as mutant alleles, nonA and nonB, was necessary for permissiveness . The gene nonA is linked very closely to rfm (cotransfer: 95%); nonB is located between dal and purB (cotransfer of nonB and purB6 : 48%) . The genetic determinant of host-specific restriction intrinsic to the Marburg strain (hsrM) was found to be identical or very closely linked to nonB . The segregation on nonB and hsrM has never been observed in the course of transduction analysis . The mutation, hsrM1, diminishes the restriction activity, but not the host-controlled modification. J Biol Chem, 1979 Feb 25, 254(4), 1080 - 9 Characterization of the extracellular lipase of Bacillus subtilis and its relationship to a membrane-bound lipase found in a mutant strain; Kennedy MB et al.; Bacillus subtilis CMK33 is a mutant that is more osmotically fragile than the wild type when it is converted to the protoplast form . The protoplasts of this mutant contain a membrane-bound lipase, which is not found in protoplasts of the wild type . Hydrolysis of the membrane lipid of mutant protoplasts by the lipase is the cause of their fragility . A protein found in the wild type organism specifically inhibits the lipase (Kent, C., and Lennarz, W . J . (1972) Proc . Natl . Acad . Sci . U . S . A . 69, 2793-2797) . This paper reports that cultures of both mutant and wild type cells contain an extracellular lipase which accumulates during the logarithmic phase of growth . The extracellular activity appears to be induced by a component of the growth medium . The membrane-bound lipase of the mutant has been partially purified and its properties have been compared to those of the extracellular lipase of the wild type . Their properties and sensitivity to the wild type inhibitor are similar, which suggests that the two molecules are closely related . The subcellular location of the lipase in the mutant has been investigated and compared to the location of the membrane-bound portion of the lipase inhibitor in the wild type . The lipase is located almost exclusively in the cytoplasmic membrane and not in mesosomal vesicles . In contrast, the lipase inhibitor is located in both types of membranes and is more concentrated in mesosomal vesicles . Under appropriate conditions, the appearance of new extracellular lipase activity in mutant cultures is paralleled by the loss of an equivalent amount of lipase activity from protoplasts prepared from the cells . This suggests that the membrane-bound lipase may be an intermediate in the secretion of the extracellular lipase . Because of the mutation in B . subtilis CMK33, which results in the absence of the lipase inhibitor, this intermediate can be found in protoplasts of the mutant, although it is not detectable in the wild type . Consequently, the mutant may be useful in studies of the mechanism of secretion of exoenzymes by Bacilli. Biochim Biophys Acta, 1979 Feb 20, 551(1), 122 - 8 A monolayer study of the reaction of trinitrobenzene sulphonic acid with amino phospholipids; Bishop DG et al.; The reaction of trinitrobenzene sulphonic acid with amino phospholipids, and in particular phosphatidylethanolamine has been studied by the monolayer technique . Injection of trinitrobenzene sulphonic acid under a monolayer of amino phospholipid results in an increase in surface pressure . The rate and extent of the pressure change is greatly affected by the initial surface pressure, the fatty acid composition of the lipid, and the presence of other non-reactive lipids, especially negatively charged phospholipids . The extent of the reaction was measured with 32P-labelled phospholipids isolated from Bacillus subtilis . Only about 80% of the phosphatidylethanolamine in the monolayer could be converted to its trinitrophenyl derivative . In the presence of negatively charged phospholipids such as cardiolipin or phosphatidylglycerol, a further 20% decrease in the trinitrophenylation of phosphatidylethanolamine was found . The pressure increase occurring during trinitrophenylation could also be correlated with the extent of the reaction by comparison of the force-area curves of pure phosphatidylethanolamine, its trinitrophenyl derivative and mixtures of both compounds . The data may offer an explanation for the observation that incomplete labelling of amino phospholipids frequently occurs in natural membranes and furthermore indicate that the use of chemical labelling techniques in the study of lipid asymmetry in biological membranes must be approached with great caution. Gene, 1979 Feb, 5(2), 87 - 91 A method for construction of specialized transducing phage rho 11 of Bacillus subtilis; Kawamura F et al.; DNA from a temperate phage rho 11 and chromosomal DNA of Bacillus subtilis 168 were digested with endonuclease EcoRI and then ligated with T4 polynucleotide ligase . The ligated DNA fragments were used to transform a lysogenic strain, B . subtilis spoA12 lys21 hisA1 leuA8 p11, and Lys+, His+ or Leu+ transformants were selected . The cells of each type were then mixed, grown and treated with mitomycin C; the induced phages were tested for abilities abilities to form plaques and to tranduce the auxotrophic marker . Various types of plaque-forming or defective phages which transduce hisA or lys marker at considerably high frequencies were thus obtained. J Gen Microbiol, 1979 Feb, 110(2), 381 - 92 Decadent sporulation mutants of Bacillus subtilis; Balassa G et al.; In decadent sporulation mutants, sporulating populations are heterogeneous: the cells reach successive chemical and physical resistances with progressively decreasing frequencies . Each decadent mutant can be characterized by the shape and slope of the curve describing the frequency of cells resistant to various agents ('the resistance spectrum') . In some mutants the resistance spectrum decreases progressively from xylene resistance to heat resistance; in other mutants it decreases rapidly between octanol resistance and chloroform resistance . Electron microscopy showed that in two mutants the majority of the cells are blocked at stages III and IV; the number of cells that develop further to reach successive morphological stages falls off progressively . In two other mutants most cells reach stage V . Cortexless spores are also frequent . One of the decadent mutations, SpoL1, was localized between aroD and acf . The phenotype of decadent mutants is discussed in terms of sequential gene activation. J Gen Microbiol, 1979 Feb, 110(2), 365 - 79 A Bacillus subtilis mutant requiring dipicolinic acid for the development of heat-resistant spores; Balassa G et al.; A Bacillus subtilis mutant is described which forms heat-resistant spores only in the presence of external dipicolinic acid (DPA) . The mutation, dpa-1, is localized in a new sporulation locus, linked to pyrA . The dpa-1 strain is unable to synthesize DPA but can incorporate external DPA . The amount of DPA incorporated, the frequency of heat-resistant spores and their degree of resistance are all dependent on the concentration of external DPA . Spores of dpa- 1 strains exhibit normal resistance to most chemicals, including octanol and chloroform, but not to ethanol, pyridine, phenol and trichloroacetic acid . Complete resistance to the latter group depends on DPA . DPA incorporation is slow and apparently requires an energy supply but not protein synthesis . Direct involvement of DPA in the heat-resistance of the spores is suggested . Thin sections of DPA-less spores exhibit clearly visible cytoplasmic membranes and ribosomes . These structures are absent or less visible in the core of spores obtained with added DPA. J Gen Microbiol, 1979 Feb, 110(2), 351 - 63 Mutants of Bacillus subtilis affected in spore outgrowth; Albertini AM et al.; Six mutants of Bacillus subtilis 168 that are temperature-sensitive in spore outgrowth were isolated . The outgrowth process proceeds normally at 35 degrees C, but at the non-permissive temperature (47 degrees C) it is arrested at a specific stage characteristic for each mutant strain . The mutants are not altered in vegetative growth whether at 35 degrees C or at 47 degrees C . They were characterized for their ability to synthesize RNA, proteins and DNA during outgrowth . A mutant defective in spore germination was also isolated; less than 5% of its spores can germinate at any of the temperatures tested . The mutations were mapped by means of transduction and transformation . The isolation of a number of outgrowth mutants which map at different loci and which affect outgrowth at different times is discussed in relation to the regulation of this process. J Antibiot (Tokyo), 1979 Feb, 32(2), 136 - 40 Preparation and antibacterial activity upon Micrococcus luteus of derivatives of iturin A, mycosubtilin and bacillomycin L, antibiotics from Bacillus subtilis; Peypoux F et al.; Methylated and acetylated derivatives of iturin A and mycosubtilin and methylated derivatives of bacillomycin L were prepared and their antibacterial activity on Micrococcus luteus was compared with the activity of the original substance . the results obtained show the importance of polar groups for the antibiotic activity of the substances of iturin group. Biokhimiia, 1979 Feb, 44(2), 332 - 7 {Effect of pancreatic DNAse on DNA synthesis in Bacillus subtilis}; Kupriianova FG et al.; An addition of pancreatic DNAse to the cultural medium is found to stimulate DNA synthesis and proliferation of Bacillus subtilis cells . Pancreatic DNAse induces a single-stranded disruption of Bac . subtilis DNA, which may act as a mechanism of DNA synthesis increase and of the culture growth acceleration. J Virol, 1979 Feb, 29(2), 540 - 5 Order of assembly of the lower collar and the tail proteins of Bacillus subtilis bacteriophage phi 29; Camacho A et al.; Extracts obtained after restrictive infection of Bacillus subtilis with mutants in cistron 11 of bacteriophage phi 29 are complemented in vitro by extract donors of the lower collar protein (p11) . Purified 11- heads, containing the major capsid protein (p8), the fiber protein (p8.5), the upper collar protein (p10), and the virus DNA, can be also complemented in vitro to produce infective virus . This result suggests that 11- heads are intermediates in phage phi 29 morphogenesis . The order of assembly of the lower collar protein p11 and the tail protein p9 was determined in vitro in two complementation steps . The results obtained indicate that the lower collar protein is assembled before the tail protein. J Gen Virol, 1979 Feb, 42(2), 305 - 14 Effect of calcium ions on the infection of Bacillus subtilis by bacteriophage SF 6; Steensma HY et al.; Infection of Bacillus subtilis 168Wt by SF 6 resulted in a rapid reduction in the number of phages . This could be counteracted by the addition of calcium, barium or strontium ions . At the optimum concentration of 7.5 x 10(-2) M, the number of p.f.u . remained constant until lysis began . Although cultures of another host . B . subtilis 31 try- his-, at the end of the logarithmic growth phase produced a substance which inactivated free phages, this was not the major cause of the reduction in the numbers of p.f.u . during infection experiments at low Ca2+ concentrations . The diminution of the number of p.f.u . was therefore attributed to the fact that at least one of the steps of the lytic cycle was calcium dependent . Adsorption of SF 6 was equally effective in media containing high or low concentrations of calcium ions . Infection experiments with phages whose DNA had been labelled radioactively revealed that, at high concentrations of calcium ions, the label remained associated with the host cells until lysis commenced . At low concentrations, however, a dissociation between phage DNA and the host was found, although adsorption took place at a normal rate . From these experiments we concluded that a high concentration of calcium ions was required for the penetration of phage DNA . Similar experiments with phages whose protein coat had been labelled showed the same results, indicating that desorption of the inactivated phages occurred . Both electron microscopy and column chromatogarphy with hydroxyapatite showed that a considerable fraction of the inactivated phages had ejected their DNA into the medium . A hypothesis explaining these results is presented. J Bacteriol, 1979 Feb, 137(2), 1028 - 30 Increased levels of dihydrofolate reductase in rifampin-resistant mutants of Bacillus subtilis; Kane JF et al.; Several independent, spontaneous rifampin-resistant mutants of Bacillus subtilis were isolated and found to have an increased resistance to trimethoprim, an inhibitor of dihydrofolate reductase . This increased resistance in the rif mutants was the result of a specific threefold increase in the activity of dihydrofolate reductase, since six other enzymes examined remained unchanged . This increased level of dihydrofolate reductase and the trimethoprim resistance were cotransformed (100%) with the rif marker . These results suggest that the RNA polymerase is altered in its recognition of the gene that specifies dihydrofolate reductase. J Bacteriol, 1979 Feb, 137(2), 933 - 46 Possible involvement of bacterial autolytic enzymes in flagellar morphogenesis; Fein JE; Autolytic enzymes were found to be required for flagellar morphogenesis in Bacillus subtilis 168 and Bacillus licheniformis 6346 . Two previously characterized, poorly lytic, chain-forming mutants of B . subtilis 168, strains FJ3 (temperature conditional) and FJ6, each 90 to 95% deficient in the production of N-acetylmuramyl-L-alanine amidase and endo-beta-N-acetylglucosaminidase, were observed to be nonmotile at 35 degrees C in a variety of liquid and semisolid meida . In contrast, cells of the isogenic wild-type strain were motile and fully separated . Electron microscopy revealed the complete absence of flagella on the mutant cells . Similar observations were made with another poorly lytic strain of B . subtilis 168 (Nil5) and with two poorly lytic, phosphoglucomutase-deficient mutants of B . licheniformis 6346 (MH-3, MH-5) . In minimal media lacking galactose (restrictive conditions), the B . licheniformis mutants failed to form flagella, or had serious abnormalities in flagellar morphogenesis and motility . Under permissive conditions, mutants FJ3 (grown at 17 degrees C) and MH-5 (grown with addend galactose) showed increased autolytic activities, grew in the dechained form, and regained their capacities to synthesize functional flagella . Examination of several classes of spontaneous revertants derived from the various mutant strains further demonstrated a close relationship between autolysin acttivity and flagellation in the two Bacillus spp. J Bacteriol, 1979 Feb, 137(2), 773 - 8 Purification and properties of the manganese-dependent phosphoglycerate mutase of Bacillus subtilis; Watabe K et al.; Phosphoglycerate mutase of Bacillus subtilis was purified to apparent homogeneity . It specifically required manganese ions for stability and activity, but it does not need 2,3-diphosphoglycerate as cofactor; the Km for Mn2+ is about 4.5 micrometer . Enzyme activity was inhibited by heavy-metal ions, 2,3-butanedione, and sulfhydryl agents . The mutase has a molecular weight of about 74,000 as shown by Sephadex gel filtration and by acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate; it consisted of one polypeptide. Mol Gen Genet, 1979 Jan 10, 168(2), 165 - 72 The genome of Bacillus subtilis phage SPP1: the arrangement of restriction endonuclease generated fragments; Ratcliff SW et al.; SPP1 DNA was cleaved by the restriction endonucleases, BglI, BglII, EcoRI, KpnI, SmaI, and SalI . The molecular weights of the DNA fragments obtained by single enzyme digestion or by consecutive digestion with two enzymes were determined by electron microscopic measurements of contour length and by gel electrophoresis . The major fragments from the six digests could be ordered to give a consistent restriction map of SPP1 . The electropherograms of several digests indicated that certain fragments occurred in less than stoichiometric amounts or were heterogeneous in size . Such bands carried a major part of radioactivity, when SPP1 DNA was terminally labelled with P32 prior to degradation by restriction enzymes . These results, and studies of the effect of exonuclease III treatment on restriction enzyme patterns define the terminal restriction fragments . All data obtained support the conclusion drawn in the preceding paper (Morelli et al., 1978 b) that the SPP1 genome is terminally redundant and partially circularly permuted. Biochemistry, 1979 Jan 9, 18(1), 198 - 202 Extracellular labeling of growing secreted polypeptide chains in Bacillus subtilis with diazoiodosulfanilic acid; Smith WP et al.; Studies of the mechanism of protein secretion in a Gram-positive bacterium, Bacillus subtilis, yielded results very similar to those previously obtained with a Gram-negative organism: nascent chains protruding from protoplasts could be labeled extracellularly; the labeled chains could be recovered on polysomes isolated from the membrane--polysome fraction; they could be released by puromycin, low Mg2+, or chain completion; the completed chains include a known secreted protein (alpha-amylase); and their ribosomes appear to be attached to membrane solely by their nascent chains . The reagent used for extracellular labeling, {1252}diazoiodosulfanilic acid, yielded severalfold more specific labeling of the nascent chains (7--10% of the total cellular labeling and one-fourth to one-third of that of the membrane--polysome fraction) than was obtained earlier with another nonpenetrating reagent. Mol Gen Genet, 1979 Jan 5, 168(1), 111 - 5 High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA; Chang S et al.; A highly efficient method for transformation of Bacillus subtilis by plasmid DNA is reported . The procedure, which involves polyethylene glycol-induced DNA uptake by protoplasts and subsequent regeneration of the bacterial cell wall, yields up to 80% transformants with an efficiency of 4 x 10(7) transformants per microgram of supercoiled DNA . Plasmids constructed by in vitro ligation or endonuclease-generated fragments of linear plasmid DNA can also transform PEG-treated protoplasts, but at a lower frequency. Mol Gen Genet, 1979 Jan 2, 167(3), 251 - 8 Characterization of plasmid transformation in Bacillus subtilis: kinetic properties and the effect of DNA conformation; Contente S et al.; Transformation of competent cells of Bacillus subtilis with antibiotic resistance plasmid DNA has shown that (a) competence for plasmid and chromosomal DNA develops with similar kinetics; (b) DNA linearized with a variety of restriction endonucleases does not transform; (c) CCC plasmid DNA is inactivated for transformation by a single nick; (d) T4 ligase restores transforming activity to both nicked and linearized DNA; (E) CCC relaxed DNA is fully active in transformation; (f) the DNA concentration-dependence of plasmid transformation is first order; and (g) plasmid transformation proceeds with a low efficiency, requiring the uptake of 10(3) to 10(4) DNA molecules per transformant . Based on this information, a model for the processing of chromosomal, plasmid and transfecting DNA is proposed. Pharmazie, 1979, 34(9), 548 - 51 {Interactions between macromolecular adjuvants and drugs . Part 18: The binding behaviour of sodium carboxymethylcellulose and other macromolecules towards streptomycin sulphate (author's transl)}; Keipert S et al.; Unlike the other macromolecules under investigation (polyvinylpyrrolidone, methylcellulose, polyvinyl alcohol), sodium carboxymethylcellulose acts on streptomycin sulphate, which leads to isolable precipitates . By equilibrium dialysis in electrolyte-free and electrolyte-containing solutions, the interaction was characterized in more detail and described by Donnan curves . The associates (which contained, on an average, 40% of streptomycin) proved to be resistant to light and air, and stable to thin-layer chromatography . Microbiological assays on the test strain Bacillus subtilis SG119 revealed a decrease in activity of the streptomycin-sodium carboxymethylcellulose associates compared to pure streptomycin (tested in the solid form, using rice granulate as a carrier substance) . In contrast, the associate was as efficient as the antibiotic when dissolved in a concentrated solution of sodium chloride. Folia Microbiol (Praha), 1979, 24(6), 462 - 72 A psychrophilic strain Actinoplanes sp . 220; Ricicova A et al.; The strain designated Actinoplanes sp . 220 differed in its characteristics from other strains of the genus Actinoplanes listed in Bergey's Manual (1974) . The strain belongs to psychrophilic culture growing within the range of 0-30 degrees C . The optimal temperature for growth on yeast--malt agar is 10-23 degrees C . Cultures transferred at 23 and 28 degrees C differed in morphological and physiological properties, enzyme activity and pigmentation in standard media . Submerged culture transferred at 28 degrees C inhibited growth of Bacillus subtilis ATCC 6633 and ATCC 9945 . LL-2,6-Diaminopimelic acid was chromatographically detected in the submerged mycelium of this culture . This compound was not found in the mycelium of the original culture transferred at 23 degrees C . The cultures did not substantially differ in the composition of other amino acids contained in larger quantities in the mycelium. Microbios, 1979, 24(96), 113 - 22 Measurement of pyrimidine dimers in spheroplasts of Bacillus subtilis; Hadden CT; A method is described for making spheroplasts of Bacillus subtilis which are permeable to exogenous enzymes . Conditions are described for measuring small numbers of pyrimidine dimers in the DNA of UV-irradiated cells by use of a partially purified Micrococcus luteus extract containing an enzyme specific for pyrimidine dimers . The system will detect as few as 10-12 pyrimidine dimers per genome. J Bacteriol, 1979 Jan, 137(1), 82 - 91 Characterization of pyrimidine-repressible and arginine-repressible carbamyl phosphate synthetases from Bacillus subtilis; Paulus TJ et al.; The number and properties of carbamyl phosphate synthetases in Bacillus subtilis have been uncertain because of conflicting genetic results and instability of the enzyme in extracts . The discovery of a previously unrecognized requirement of B . subtilis carbamyl phosphate synthetases for a high concentration of potassium ions for activity and stability permitted unequivocal demonstration that this bacterium elaborates two carbamyl phosphate synthetases . Carbamyl phosphate synthetase A was shown to be repressed by arginine, to have a molecular weight of about 200,000, and to be coded for by a gene that maps near argC4 . This isozyme was insensitive to metabolites of the arginine and pyrimidine biosynthetic pathways . Carbamyl phosphate synthetase P was found to be repressed by uracil, to have a molecular weight of 90,000 to 100,000, and to be coded for by a gene that maps near the other pyr genes . This isozyme was activated by phosphoridine nucleotides . Other kinetic properties of the two isozymes were compared . Bacillus thus resembles eucaryotic microbes in producing two carbamyl phosphate synthetases, rather than the enteric bacteria, which produce a single carbamyl phosphate synthetase. Zentralbl Bakteriol Naturwiss, 1979, 134(4), 352 - 9 Subtilopeptidase A produced by Bacillus subtilis PR-70 . II . Kinetic behaviour of immobilized enzyme; Attia RM et al.; The kinetic behaviour of immobilized subtilopeptidase A was investigated . The enzyme was obtained from a local isolate of B . subtilis PR-70 . Using different inorganic supports, Amberlite CG-50 was superior in this respect . It gave 97.8% adsorption, followed by silica gel GC . The values of K and K2 for the rate of enzyme catalyzed being 8.75 and 2.06, respectively . The behaviour of v against Et is the same as v against St . Michaelis' constant was determined using different methods . The average of Km value and Vmax were 0.0094 and 0.95, respectively . Studying how v behaves when St is varied while Et is constant, two active site per enzyme molecule and auto-inhibition of enzyme by its own substrate were observed . Comparing kinetic parameters of a soluble and insoluble subtilopeptidase A showed that Km decreased from 0.016 to 0.0094, while Vmax increased from 0.71 to 0.95, respectively . This indicated that when subtilopeptidase was bound to Amb . GC-50, a case of partially non-competitive inhibition occurred . The recovery of enzymatic activity in the water insoluble subtilopeptidase A is 12.8 per cent. Microbiol Immunol, 1979, 23(8), 727 - 34 Growth and sporulation of auxotrophs of Bacillus subtilis in a medium with limited nutrients; Nishihara T; Growth and sporulation were examined for 30 auxotrophs of Bacillus subtilis in a chemically defined medium with suboptimal amounts of nutrients . All strains except for some adenine-requiring mutants could not overtake sporulation stage II when amino acids, vitamins, or bases were limited, whereas they sporulated fairly well without limitation . Abnormal structures, a cell with thickened cell wall and a cell with several refractile bodies, were found in some strains after the vegetative growth stopped. Mol Gen Genet, 1979, 177(1), 23 - 9 Chromosomal mutations causing resistance to tetracycline in Bacillus subtilis; Williams G et al.; We have isolated, after ethylmethanesulfonate mutagenesis, several chromosomal mutations causing resistance to tetracycline in Bacillus subtilis . These mutations fall into two classics, tetA and tetB . 30 S ribosomal protein S10 shows an altered mobility on two-dimensional acrylamide gels in cells bearing the former type of mutation . Ribosomes from these cells show elevated levels of resistance to tetracycline in vitro as measured by polyuridine dependent polyphenylalanine synthesis . The tetA locus maps adjacent to the tuf gene in the B . subtilis ribosomal protein gene cluster . Cells with the tetB mutation do not show any altered ribosomal protein, and their ribosomes are as sensitive, in vitro, to tetracycline as ribosomes isolated from wild type cells . The tetB mutation has been mapped proximal to cysA14. Ultramicroscopy, 1979, 4(4), 395 - 412 Low-dose electron image reconstruction of negatively stained contractile phage sheath from Bacillus subtilis (PBS-Z); Cremers AF et al.; The structure of the contractile sheath of the defective phage from B . subtilis (PBS-Z) has been investigated by low-dose electron microscopy and image reconstruction . The extended and contracted sheath particles were imaged by means of two negative stains which consisted of uranyl- and phosphotungstate-containing solutions of a pH of 4.2 and 7.0 respectively . Images of identical parts of the same type of specimen were recorded at a total electron dose of 80 C/m2 (5 electrons/A2) and 4 x 10(3) C/m2 (250 electrons/A2) . The low-dose reconstructions of the extended and contracted sheath structure in the two stains show good correspondence and made it possible to draw the following structural conclusions . The sheath protein in both types of structure has an elongated shape, and in both structures the long molecular axis lies in a plane perpendicular to the helical sheath axis . The orientation of the protein in the extended and contracted sheath is different; the long axes differ by about 35 degrees in orientation . The reconstructions did not permit conclusions about different conformational states of the protein in both structures . These data, together with the packing parameters of the protein subunits in the contractile sheath {1}, form the complete structural analysis of this biological structure by electron microscopy . The radiation damage effects which have been monitored in analyzing image pairs to the full extent may be summarized as follows . (1) Diameters of the sheath structure increase, which indicate flattening . (2) There is no loss in resolution, and layerline altitudes of the Fourier-transformed images do not change . (3) Uranyl stain behaves differently compared to phosphotungstate . In both negative stains the structural noise level increases upon irradiation as follows from the increase in phase residuals of the digital layerline data . In uranyl-stained images also more aperiodic noise appears . (4) The Fourier amplitudes of the principal layerline maxima shift towards lower spatial frequencies; phases of corresponding maxima generally remain constant . This pattern is more pronounced in the extended sheath data; there is no rationale describing these positional shifts . Moreover, in the case of contracted sheath the amplitudes of Fourier components also change more in absolute value . Therefore the damage effects also seem to depend on the type of structure embedded in the stain . (5) In the reconstructed images these radiation effects create artificial stain-excluded volumes of a type and at a radius which depend on the stain and structure. Microbios, 1979, 24(95), 29 - 39 Comparison of three procedures for isolating DNA from bacteria; Amundesn SK et al.; Three methods employing chloroform-isoamyl alcohol (CI), phenol, or enzymes, were evaluated for isolating DNA from Escherichia coli, Bacillus subtilis, and Arthrobacter globiformis . For the amounts of reagents employed at optimum conditions in the CI and phenol procedures, 0.4-0.9 mg of DNA/g wet weight of cells was isolated . Using the enzymatic procedure, approximately twice as much DNA was isolated . DNA isolated by the CI procedure contained 0.03-0.09% protein and 0.08-0.12% RNA . DNA isolated by the phenol procedure contained 0.02-0.05% protein and 2.2-2.6% RNA . DNA isolated by an enzymatic procedure, which is described in detail, contained 32.2-45.7% protein and 0.3-0.6% RNA . DNA isolated by all three procedures are double-stranded and at least 10(6) in molecular weight, as suggested by data from thermal transition analyses and transformations . These data emphasize that the desired characteristics of DNA for experimental purposes must be considered in selecting an isolation procedure. Folia Microbiol (Praha), 1979, 24(5), 373 - 5 Exoprotease production by sporogenous and asporogenous mycobacillin non-producer mutants of Bacillus subtilis; Bose R et al.; Mycobacillin non-producers, whether sporogenous or asporogenous, possess less exoprotease, but effective exoprotease producers are not always good mycobacillin yielders . There might exist a minimum level of exoprotease formation for elaboration of mycobacillin. Genetika, 1979, 15(3), 594 - 604 {Production and study of Bacillus subtilis mutants for genes involved in nucleoside catabolism}; Rumiantseva EV et al.; By means of selection for a low thymine requirement the mutants fo thymine auxotrophs for deoxyriboaldolase (dra) and phosphodeoxyribomutase (drm) genes were obtained . Besides the mutants for pyrimidinenucleoside phosphorylase gene (pdp) were olso isolated using selection on the fluorodeoxyuridine resistance . The latter enzyme provides for pyrimidine nucleosides catabolism (thymidine, uridine) in Bacilli, as well as the conversion of exogenous thymine to thymidine in thymine auxotrophs . The data obtained when studying the deo-enzymes activities in various types of the mutants and also under the condition of induction by thymidine and acetoaldehyde are in accordance with the assumption that deoxyriboso-5-phosphate is an inductor of the deo-enzymes in Bacillus subtilis . The genes dra and pdp were tightly linked as it had been shown by the transformation experiments; in contrast, no linkage was revealed between dra and drm or pdp and drm . A secondary mutation (adn), not linked with dra and blocking the ability of bacteria to catabolise adenosine (purine nucleoside phosphorylase activity remains constant) was found in some dra-mutants. Zentralbl Bakteriol Naturwiss, 1979, 134(3), 275 - 81 Subtilopeptidase A produced by Bacillus subtilis PR-70 . I . Kinetic behaviour of solubilized enzymes; Attia RM et al.; The kinetic behaviour of subtilopeptidase A was investigated . The enzyme was obtained from a local isolate of B . subtilis PR-70 . The rate of enzyme catalyzed conversion of substrate to product is directly proportional to the enzyme concentration, v = K(E) . Michaelis constant was determined using different methods . The average of Km value is equal to 0.01615 . The Vmax and Et were determined being 0.71 and 1.467, respectively, using KUNITZ's casein digestion method . The enzymeconcentration involved in the reaction system is equal to 97.8% . A trial to calculate the molecular weight and the number of active groups were discussed. Biochimie, 1979, 61(3), 385 - 91 Characterization of an inhibitor of the intracellular protease from Bacillus subtilis; Millet J et al.; A specific inhibitor of intracellular serylprotease from Bacillus subtilis has been isolated from both growing and sporulating cells . Like other protease inhibitors isolated from eukaryotic cells, the inhibitor from B . subtilis is a thermostable protein . A purification method is described . The molecular weight estimated by Biogel filtration and SDS gel electrophoresis is about 15,500 . Both proteolytic and esterolytic activities of intracellular protease are equally sensitive to inhibition . With azocoll or Z-tyrosine p-nitrophenylester as substrates, noncompetitive inhibition patterns are observed . The inhibitor has no effect on the proteolytic or esterolytic activities of the extracellular serylprotease . A similar thermostable inhibitor is also present in Bacillus megaterium. Biochimie, 1979, 61(1), 93 - 100 On the nature of tetracycline resistance in Bacillus subtilis mediated by the plasmid pT 127; Fargette F et al.; The nature of tetracycline resistance was studied in a strain of Bacillus subtilis carrying the plasmid pT 127 in comparison with the parental strain . The resistance has been shown to be inducible in both strains upon exposure to subinhibitory concentrations of tetracycline . No modification of the protein-synthesizing activity of the ribosomes or intracellular inactivation of the antibiotic was observed in both strains . Accumulation of labeled tetracycline in B . subtilis was found to be particularly low in the wild-type strain, compared to other bacterial species, with concentration gradients of only 2 to 3 fold . From the kinetics obtained it is likely that the permeation of the antibiotic does not correspond to an active process in B . subtilis . A fairly good correlation was established between the level of resistance obtained after induction or by the presence of the plasmid pT 127 and a decrease in the binding capacity of the cell for the antibiotic. J Virol, 1979 Jan, 29(1), 61 - 8 Deoxythymidine nucleotide metabolism in Bacillus subtilis W23 infected with bacteriophage SP1Oc: preliminary evidence that dTMP in SP10c DNA is synthesized by a novel, bacteriophage-specific mechanism; Markewych O et al.; Despite the fact that mature SP10c DNA contains dTMP, the acid-soluble fraction of infected cells contained no dTTP during the interval of phage replication . However, infected cells contained normal cellular levels of dATP, dGTP, and dCTP . Upon infection of deoxythymidine-starved Bacillus subtilis M160 (a deoxythymidine-requiring mutant of B . subtilis W23), mature phage DNA with a normal dTMP content was made . SP10c codes for an enzyme that seems to catalyze the tetrahydrofolate-dependent transfer of 1-carbon fragments to the 5 position of dUMP . The transfer of 1-carbon fragments is not accompanied by oxidation of tetrahydrofolage to dihydrofolate, implying that the enzyme in question is not a dTMP synthetase . It is proposed that dTMP in mature SP10c DNA is derived by the postreplicational modification of some other nucleotide and not by the direct incorporation of dTTP into DNA. Gene, 1979 Jan, 5(1), 1 - 7 Unusual base sequence arrangement in phage phi 29 DNA; Ito J et al.; Susceptibility of Bacillus subtilis phage phi 29 DNA to 34 different restriction endoculceases was determined . Three enzymes, BglI, XbaI and BstEII, were found to cleave phi 29 DNA only once at specific sites . The sites of these single cleavages have been mapped . Thirteen enzymes did not cut phi 29 DNA . phi 29 HindIII DNA fragments inserted into pBR313 plasmid and propagated in Escherichia coli, were resistant to these restriction endonucleases . This result suggests that the insusceptibility is due to the absence of the nucleotide sequences on phi 29 recognized by the enzymes, and not to the presence of modified nucleotides. Mikrobiologiia, 1979 Jan-Feb, 48(1), 93 - 8 {Experimental verification of the results of the mathematical modeling of the process of formation of alpha-amylase-nonproductive mutants of Bacillus subtilis under continous cultivation}; Fencl Z et al.; When Bacillus subtilis produces alpha-amylase in the course of continuous cultivation, it is difficult to maintain the activity at a constant level . This may be due to the formation of nonproductive mutants . Individual cells in the population have been analysed in the course of the continuous process . The composition of the population changes depending on time and the composition of the growth medium . Semisynthetic media cause selection of mutants which synthesize the enzyme at a low rate . In contrast, complex media which are more enriched in the sources of carbon and nitrogen induce accumulation of mutants with a high activity. J Bacteriol, 1979 Jan, 137(1), 635 - 43 Plasmid copy number control: isolation and characterization of high-copy-number mutants of plasmid pE194; Weisblum B et al.; A plasmid, pE194, obtained from Staphylococcus aureus confers resistance to macrolide, lincosamide, and streptogramin type B ("MLS") antibiotics . For full expression, the resistance phenotype requires a period of induction by subinhibitory concentrations of erythromycin . A copy number in the range of 10 to 25 copies per cell is maintained during cultivation at 32 degrees C . It is possible to transfer pE194 to Bacillus subtilis by transformation . In B . subtilis, the plasmid is maintained at a copy number of approximately 10 per cell at 37 degrees C, and resistance is inducible . Tylosin, a macrolide antibiotic which resembles erythromycin structurally and to which erythromycin induces resistance, lacks inducing activity . Two types of plasmid mutants were obtained and characterized after selection on medium containing 10 microgram of tylosin per ml . One mutant class appeared to express resistance constitutively and maintained a copy number indistinguishable from that of the parent plasmid . The other mutant type had a 5- to 10-fold-elevated plasmid copy number (i.e., 50 to 100 copies per cell) and expressed resistance inducibly . Both classes of tylosin-resistant mutants were shown to be due to alterations in the plasmid and not to modifications of the host genome. J Bacteriol, 1979 Jan, 137(1), 391 - 6 DNA repair in Bacillus subtilis: excision repair capacity of competent cells; Yasbin RE et al.; Competent Bacillus subtilis were investigated for their ability to support the repair of UV-irradiated bacteriophage and bacteriophage DNA . UV-irradiated bacteriophage DNA cannot be repaired to the same level as UV-irradiated bacteriophage, suggesting a deficiency in the ability of competent cells to repair UV damage . However, competent cells were as repair proficient as noncompetent cells in their ability to repair irradiated bacteriophage in marker rescue experiments . The increased sensitivity of irradiated DNA is shown to be due to the inability of excision repair to function on transfecting DNA in competent bacteria . Furthermore, competent cells show no evidence of possessing an inducible BsuR restriction system to complement their inducible BsuR modification enzyme. J Bacteriol, 1979 Jan, 137(1), 35 - 43 Cell wall teichoic acid as a reserve phosphate source in Bacillus subtilis; Grant WD; Although exponential growth of Bacillus subtilis 168 in a phosphate-limited medium halted with the exhaustion of inorganic phosphate, the bacteria continued to grow at a slower rate for a further 3 to 4 h at 37 degrees C . This postexponential growth in the absence of an exogenous phosphate supply was accompanied by a loss of teichoic acid from the cell walls of the bacteria . Quantitative analysis of walls and culture fluids showed that the phosphate loss from the walls could not be accounted for by an increase in phosphate-containing compounds in the medium, which implied that the cells were using their own wall teichoic acids to supply phosphate necessary for growth . Addition of exogenous teichoic acid to phosphate-starved cultures resulted in stimulation of growth and in the simultaneous disappearance of teichoic acid phosphate from the medium . It is proposed that teichoic acids, which can contain more than 30% of the total phosphorus of exponential-phase cells, can be used as a reserve phosphate source when the bacteria are starved for inorganic phosphate. J Bacteriol, 1979 Jan, 137(1), 327 - 31 Control of teichoic acid synthesis during phosphate limitation; Glaser L et al.; The synthesis of teichoic acids was examined in Bacillus subtilis Marburg grown under conditions of phosphate limitation . The results indicate that the inhibition of polyglycerolphosphate synthesis observed under these conditions is the result of two processes . The first process is reversible and is independent of new protein synthesis; the second process is irreversible and requires the synthesis of new protein . During growth, under conditions of phosphate limitation, there is a slow decrease in the level of CDP glycerol pyrophosphorylase activity which is by itself not sufficient to account for the decrease in the rate of polyglycerolphosphate synthesis. J Bacteriol, 1979 Jan, 137(1), 213 - 20 Inhibition of Bacillus subtilis growth and sporulation by threonine; Lamb DH et al.; A 1-mg/ml amount of threonine (8.4 mM) inhibited growth and sporulation of Bacillus subtilis 168 . Inhibition of sporulation was efficiently reversed by valine and less efficiently by pyruvate, arginine, glutamine, and isoleucine . Inhibition of vegetative growth was reversed by asparate and glutamate as well as by valine, arginine, or glutamine . Cells in minimal growth medium were inhibited only transiently by very high concentrations of threonine, whereas inhibition of sporulation was permanent . Addition of threonine prevented the normal increase in alkaline phosphatase and reduced the production of extracellular protease by about 50%, suggesting that threonine blocked the sporulation process relatively early . 2-Ketobutyrate was able to mimic the effect of threonine on sporulation . Sporulation in a strain selected for resistance to azaleucine was partially resistant . Seventy-five percent of the mutants selected for the ability to grow vegetatively in the presence of high threonine concentrations were found to be simultaneously isoleucine auxotrophs . In at least one of these mutants, the threonine resistance phenotpye could not be dissociated from the isoleucine requirement by transformation . This mutation was closely linked to a known ilvA mutation (recombination index, 0.16) . This strain also had reduced intracellular threonine deaminase activity . These results suggest that threonine inhibits B . subtilis by causing valine starvation. J Bacteriol, 1979 Jan, 137(1), 189 - 96 Carbon and nitrogen repression of arginine catabolic enzymes in Bacillus subtilis; Baumberg S et al.; Specific activities of arginase and ornithine aminotransferase, inducible enzymes of arginine catabolism in Bacillus subtilis 168, were examined in cells grown with various carbon and nitrogen sources . Levels of these enzymes were similar in arginine-induced cultures whether glucose or citrate was the carbon source (in contrast to histidase), suggesting that carbon source catabolite repression has only limited effect . In media with combinations of nitrogen sources, glutamine strongly repressed induction of these enzymes by proline or arginine . Ammonium, however, only repressed induction by proline and had no effect on induction by arginine . These effects correlate with generation times in media containing these substances as sole nitrogen sources: growth rates decreased in the order glutamine-arginine-ammonium-proline . Similar phenomena were observed when glutamine or ammonium were added to arginine- or proline-grown cultures, or when arginine or proline were added to glutamine- or ammonium-grown cultures . In the latter cases, an additional feature was apparent, namely a surprisingly long transition between steady-state enzyme levels . The results are compared with those for other bacteria and for eucaryotic microorganisms. J Bacteriol, 1979 Jan, 137(1), 124 - 8 Effect of 6-(p-hydroxyphenylazo)-uracil on the homologous and heterologous transduction processes in Bacillus subtilis; Canosi U et al.; We have studied the effect of 6-(p-hydroxyphenylazo)-uracil on the recombination processes that operate in the homologous and heterologous transduction mediated by PBS1 and SP10 phages of Bacillus subtilis . The results obtained demonstrate that the process of heterologous genetic exchange is sensitive to this compound, whereas the homologous process is not . The present data, along with those of our previous work (U . Canosi, A . G . Siccardi, A . Falaschi, and G . Mazza, J . Bacteriol . 126:108--121, 1976), suggest that the DNA polymerase III is involved in the recombination process that operates in transformation and heterologous transduction, whereas homologous transduction follows a partially independent pathway not involving this protein. Prikl Biokhim Mikrobiol, 1979 Jan-Feb, 15(1), 57 - 62 {Effect of sources of carbon, nitrogen, and phosphorus on the synthesis of proteases from Bacillus subtilis cultures}; Kaluniants KA et al.; The paper gives data on the influence of different sources of carbon, nitrogen and phosphorus on the accumulation of biomass and synthesis of neutral protease by Bacillus subtilis str . 103 . The highest proteolytic activity was obtained on the medium containing maltose or hydrolyzed starch as a carbon source, monopotassium phosphate as a phosphorus source, and ammonium sulphate as a nitrogen source . The enzyme activity increased upon an addition of albumin, peptone or casein . A study of the amino acid effect showed that methionine, isoleucine and valine stimulated the protease synthesis to the greatest extent. Biochim Biophys Acta, 1978 Dec 21, 521(2), 719 - 25 Asymmetric transcription during post-germinative development of Bacillus subtilis spores . II . Hybrid competition analyses; Setoguchi Y et al.; Hybrid-competition analyses were done to estimate the relatedness of 3H-labeled mRNA species synthesized during spore germination and log-phase growth . The competitions showed that early in the germination process 10--15 and 1--3% of the RNA transcribed from the H and from the L strand, respectively, were unique and absent during log-phase growth . At later stages, the amounts of the germination-specific H transcripts decreased more rapidly than the L transcripts . The competitions with pulse-labeled log-phase RNAs showed that vegetative genes were transcribed more rapidly from the H strand than from the L strand . Most of the results could be correlated with the observed decrease in the H/L asymmetry ration during spore germination. Biochim Biophys Acta, 1978 Dec 21, 521(2), 708 - 18 Asymmetric transcription during post-germinative development of Bacillus subtilis spores; Margulies L et al.; The relative transcription from L and H strands of Bacillus subtilis DNA during consecutive stages of spore outgrowth was determined and compared to the transcription pattern during log-phase growth of vegetative cells . Pulses of {3H} uridine were administered during early, middle and late outgrowth phases of germination and the RNAs isolated . The asymmetry ratio of H/L as determined by hybridization at saturating RNA/DNA inputs showed a gradual decrease . During the period studied (10-90 and 90-160 min post-induction), about 50 and 35%, respectively, of the radioactive RNA consisted of ribosomal RNA transcripts . The decrease in the H/L asymmetry ratio was due predominantly to the appearance and accumulation of L strand transcripts and not to either changes in the quantity of H strand transcripts nor to fluctuation in the rate of rRNA synthesis. Biochim Biophys Acta, 1978 Dec 21, 521(2), 484 - 92 Transformation in Bacillus subtilis with nitrogen mustard crosslinked DNA . Effect on cotransformation and mutation frequencies; Scheinbach S et al.; Bacillus subtilis DNA was treated in vitro with nitrogen mustard and the crosslinked molecules were purified, after alkali denaturation, by hydroxyapatite chromatography . When tested for the ability to transform the trpC2-hisB2 segment, these molecules exhibited a decrease in the cotransformation index (r) as compared to native or renatured DNA . The decrease in r was not accompanied by an increase in mutagenicity. Arch Microbiol, 1978 Dec 20, 119(3), 287 - 93 On the appearance of Bacillus subtilis intracellular serine protease in the cell membrane and culture medium . Comparison of the enzyme and other Bacillus subtilis serine proteases; Strongin AY et al.; While about 80% of the cell-bound intracellular serine protease of Bacillus subtilis A-50 have been recovered in the soluble fraction upon disruption of cells, the rest of the enzyme was found to be associated with the membrane fraction . Soluble cytoplasmic intracellular serine protease, as well as membrane-bound serine protease liberated by non-ionic detergent treatment, have been isolated in a pure state and shown to be identical . The same protease might also be found extracellularly, due presumably to cell lysis or altered membrane permeability . Intracellular serine protease of Bacillus subtilis A-50 was clearly related to Bacillus subtilis serine proteases W1 and bacillopeptidase F described as extracellular enzymes. Biochem J, 1978 Dec 15, 176(3), 639 - 47 Inhibition of bacterial transport by uncouplers of oxidative phosphorylation . Effects of pentachlorophenol and analogues in Bacillus subtilis; Nicholas RA et al.; Analogues of the potent uncoupler of oxidative phosphorylation pentachlorophenol were tested as inhibitors of proline and glycine transport by Bacillus subtilis . These analogues included less highly substituted chlorophenols and pentachlorothiophenol . Like pentachlorophenol, they are non-competitive inhibitors of proline transport and uncompetitive inhibitors of glycine transport . However, the less highly substituted chlorophenols are weaker acids than pentachlorophenol and also weaker inhibitors . Analysis indicated that the anionic form of the uncouplers is the inhibiting species . Pentachlorothiophenol, a water-insoluble anion, is also a potent inhibitor . These results support previous studies that concluded that uncouplers of oxidative phosphorylation inhibit amino acid transport by binding at specific sites on proteins, the free energy of interaction stabilizing 'unproductive' conformations . Such specific interactions of uncoupler with protein are probably commonplace. J Biol Chem, 1978 Dec 10, 253(23), 8559 - 63 Purification and properties of a Bacillus subtilis endonuclease specific for apurinic sites in DNA; Inoue T et al.; An endonuclease which hydrolyzes depurinated DNA has been purified from extracts of Bacillus subtilis cells . The endonuclease is a monomeric protein and has a molecular weight of around 56,000 . The enzyme is specific for apurinic sites in double-stranded DNA, has a pH optimum at 8.0, and is slightly stimulated with 50 mM NaCl but completely inhibited with 500 mM NaCl . It requires no divalent cations and is insensitive to EDTA; it has no associated exonuclease . These properties are very similar to those of Escherichia coli endonuclease IV, which is also insensitive to EDTA and has no exonuclease activity, and very different from those of the main endonuclease for apurinic sites (endonuclease IV) of the same bacterium. J Biol Chem, 1978 Dec 10, 253(23), 8518 - 25 Bacillus subtilis bacteriophage PBS2-induced DNA polymerase . Its purification and assay characteristics; Hitzeman RA et al.; The DNA polymerase induced by Bacillus subtilis bacteriophage PBS2 (whose DNA contains uracil instead of thymine) has been purified and characterized for its specificity . The enzyme requires a high ionic strength for optimal stability and activity and is sensitive to various anions and to sulfhdryl reagents . Both dUTP and dTTP are incorporated efficiently as substrates and are competitive inhibitors at the same active site . The apparent Km and Ki values are about 6 micrometers for dTTP and 15 micrometers for dUTP, when denatured, uracil-containing B . subtilis or salmon sperm DNA (3.9 micrometers for dUTP and 2.6 micrometers for dTTP) . The PBS2 enzyme works best on denatured DNA, on double-stranded DNA activated by DNase to produce gaps, or on primed homopolymeric DNA . Using denatured DNA preparations of average molecular weight 6.2 million, the apparent Km values are 270 micrograms/ml for B . subtilis DNA and 360 micrograms/ml for PBS2 DNA; the Vmax value for denatured PBS2 DNA containing uracil is 7-fold greater than that for denatured B . subtilis DNA containing thymine . However, lower molecular weight DNAs have 10-fold lower apparent Km values and show similar Vmax values for both B . subtilis and PBS2 DNAs . Thus, the PBS2 phage-induced DNA polymerase (which likely replicates only uracil-containing phage DNA using dUTP in vivo) has little selectivity for uracil- versus thymine-containing deoxyribonucleotides or DNA in vitro. J Biol Chem, 1978 Dec 10, 253(23), 8526 - 32 Characterization of the Bacillus subtilis bacteriophage PBS2-induced DNA polymerase and its associated exonuclease activity; Hitzeman RA et al.; The DNA polymerase induced by Bacillus subtilis bacteriophage PBS2 has a Stokes radius of 7.2 in buffers of high ioninc strength, suggesting a molecular weight in the range 145,000 to 195,000 . The polypeptide bands observed on gel electrophoresis in dodecyl sulfate have apparent molecular weights of 78,000 and 69,000 (and possibly another 27,000) in equimolar amounts . In buffers of low ionic strength, the enzyme appears to form large aggregates and even precipitates, with about 90% loss of activity . A nuclease activity co-purifies with the PBS2 DNA polymerase and shows similar responses to changes in pH, MgCl2, N-ethylmaleimide, temperature, and dextran sulfate levels . The nuclease produces deoxyribonucleoside 5'monophosphates from denatured DNA containing thymine or uracil . No endonuclease activity is detectable on supercoiled DNA . The inhibition of nuclease activity by added deoxyribonucleoside triphosphates, the DNA-dependent turnover of triphosphates, to free monophosphates during DNA polymerization, the inhibition of nuclease activity by 3'-phosphates on the DNA template-primer, and the pattern of digestion of 5'-{32P}phosphate-labeled DNA all indicate that the PBS2 DNA polymerase-associated hydrolytic activity is a 3' leads to 5'-exonuclease. J Gen Microbiol, 1978 Dec, 109(2), 225 - 36 Linkage analysis in Dictyostelium discoideum using multiply marked tester strains: establishment of linkage group VII and the reassessment of earlier linkage data; Ratner DI et al.; To aid linkage analysis and mapping studies in Dictyostelium discoideum, we have constructed several tester strains with easily scored mutations characterizing the six currently identified linkage groups . Use has been made of conditionally lethal mutants unable to grow upon Bacillus subtilis, and the locus of the mutation involved (bsgA) has been assigned to linkage group III . The mutation cobA1, which confers resistance to cobaltous chloride, has been assigned to a previously unidentified linkage group (VII) . The temperature-sensitive growth mutation tsgC7, previously reported to define linkage group V, has been reassigned to group III, leaving linkage group V presently unmarked . The further use of genetic tester strains is described. J Virol, 1978 Dec, 28(3), 753 - 66 Production and expression of dTMP-enriched DNA of bacteriophage SP15; Casella E et al.; Normal DNA of Bacillus subtilis phage SP15 contains approximately equimolar quantities of dTMP and a hypermodified nucleotide, 5-dihydroxypentyl-dUMP (DHPdUMP) . Deoxythymidine (dThd) rescue of phage DNA synthesis in 5-fluorodeoxyuridine (FUdR)-inhibited cultures resulted in the synthesis of SP15 DNA containing enhanced levels of dTMP and correspondingly reduced levels of DHPdUMP . This rescued system was used to probe possible roles of DHPdUMP in phage development . The results suggested that normal levels of DHPdUMP were not required for proper transcription of phage DNA, but normal amounts of DHPdUMP were indispensable for phage assembly and/or DNA maturation . The amount of exogenous dThd required to rescue phage DNA synthesis in FUdR-inhibited cultures was 20-fold higher than the concentration required to rescue cellular replication, whereas the same low concentrations of dThd sufficed to rescue viral and bacterial DNA syntheses in aminopterin-inhibited cultures . Normal SP15 DNA was made in rescued, aminopterin-inhibited cultures . We suggest that FUdR (but not aminopterin) partially suppresses biosynthesis of the hypermodified nucleotide and that there is a barrier to replacement of DHPdUMP by dTMP; therefore, exceptionally large amounts of dThd must be salvaged in FUdR-inhibited cultures to force replacement of the unusual nucleotide by dTMP. Can J Microbiol, 1978 Dec, 24(12), 1439 - 51 The effect of chemical fixatives on cell walls of Bacillus subtilis; Beveridge TJ et al.; Cell walls of Bacillus subtilis were treated with several chemical fixatives which are commonly used preparatory to electron microscopy; i.e., osmium tetroxide, formaldehyde, acrolein, crotonaldehyde, and glutaraldehyde . Dimensional analysis was performed on thin sections of fixed walls from plastic embeddings and, by means of the statistical technique of multiple comparisons, significant differences were found between wall thicknesses from the various fixations . These differences varied with the fixation time and the type of fixative used in the reaction . When compared to embedded walls which had been stained before fixation, the overall effect was a reduction in wall thickness which was attributed to fixative action and not to the embedding or staining processes . The reduction of wall thickness was even more apparent when dimensions of fixed walls were compared to published dimensions of both frozen sections and freeze-etch profiles . Since these fixatives bind to reactive sites within the wall fabric, a change in electrochemical charge density is effected which can be monitored in terms of heavy-metal-binding capacity . Most monoaldehyde fixatives and osmium tetroxide render the wall as reactive, or less reactive, to uranyl acetate as unfixed walls, whereas glutaraldehyde can significantly increase the binding capacity. J Gen Virol, 1978 Dec, 41(3), 563 - 72 Studies on transduction process by SPP1 phage; Ferrari E et al.; The conditions for optimal transduction efficiency of the Bacillus subtilis phage SPP1 have been investigated . By irradiating transducing lysates with u.v . light we have been able to obtain a fivefold increase in the number of transductants and to reduce strongly the interference caused by infective particles . Any dependence of SPP1 transduction on PBSX induction has been ruled out by the use of xin mutants, which are unable to induce the defective phage . SPP1 mediated transduction is susceptible to the restriction and modification system of B . subtilis . The rec functions involved in the recombination of the SPP1 transduced DNA fragment are probably identical to those required in DNA transformation and heterologous PBS1 transduction. Biokhimiia, 1978 Dec, 43(12), 2233 - 40 {Immunochemical study of subtilisins obtained from Bacillus subtilis A-50 and some of its mutants}; Logunov AI et al.; Physico-chemical parameters of subtilisins from the original Bacillus subtilis A-50 strain (proteolytic activity, electrophoretic mobility, molecular weight, reactions with specific inhibitors) were similar to those mentioned in the literature for the enzymes of other strains . Immunological experiment has shown, that Bacillus subtilis A-50 subtilisins with various electrophoretic mobility do not differ in their antigenic properties . Enzymes with high electrophoretic mobility from mutant strains were similar to I--III subtilisin fractions from Bac . subtilis A-50 in the antigenic characteristics . However, the antigenic heterogeneity was observed in I, II and III enzyme fractions of some mutant strains . Subtilisins studied appear to form the isoenzyme system. J Antibiot (Tokyo), 1978 Dec, 31(12), 1211 - 7 CC-1065 (NSC-298223), a new antitumor antibiotic . Production, in vitro biological activity, microbiological assays and taxonomy of the producing microorganism; Hanka LJ et al.; A new antitumor antibiotic is produced in fermentation liquors of Streptomyces zelensis sp.n . The antibiotic is biologically active at extremely low concentrations . At 40 pg/ml, it inhibited 90% of the growth of L1210 cells in culture in tube dilution assays . The minimal inhibitory concentrations against Gram-positive bacteria is between 1 approximately 10 ng/ml, while these values for Gram-negative bacteria and fungi are mostly under 1 microgram/ml . A microbiological assay with Bacillus subtilis can detect concentrations of 1 approximately 2 ng/ml. Proc Natl Acad Sci U S A, 1978 Dec, 75(12), 5993 - 7 Thiostrepton-resistant mutants exhibit relaxed synthesis of RNA; Smith I et al.; Spontaneous mutants of Bacillus subtilis resistant to thiostrepton (TSP) exhibit relaxed synthesis of RNA when starved for required amino acids . Intact cells of tsp mutants cannot synthesize the regulatory nucleotides, ppGpp and pppGpp, after amino acid deprivation . Because ribosomes isolated from spontaneous revertants to thiostrepton sensitivity and from wild-type stringent strains can synthesize (p)ppGpp whereas ribosomes isolated from tsp strains cannot synthesize these regulatory nucleotides in the presence of stringent factor, it appears that the lesion is expressed at the level of the ribosome . Genetic mapping, via three-factor transformational crosses, has shown that tsp is closely linked to rif, in the order cysA14, tsp, rif-I, strA . The phenotype of the tsp mutants indicates that they are of the relC type . Their map position indicates that they are different from a previously described B subtilis rel mutation . Ribosomes from the latter strain can synthesize (p)ppGpp in cell-free extracts. Proc Natl Acad Sci U S A, 1978 Dec, 75(12), 5922 - 5 Interaction of secreted nascent chains with surrounding membrane in Bacillus subtilis; Smith WP et al.; To determine the length of secreted nascent polypeptide chain that is surrounded by membrane, we digested labeled nascent chains protruding from protoplasts of Bacillus subtilis with Pronase and isolated the residual ribosome-attached chains from the membrane-polysome fraction . Gel chromatography revealed a sharp major peak that had been protected by membrane plus bound ribosomes . The ribosomes themselves protected half as great a length . Because no free chain between the ribosome and the membrane was detected by Pronase treatment, the difference between the two protected lengths should measure the length protected by the membrane . More accurate measurements of these lengths, obtained by dansylation of the exposed NH2 terminus of the isolated fragments, yielded a difference of 21 amino acids . This value corresponds to an extended chain of 75 A, which is approximately the thickness of the bacterial cell membrane . We earlier presented evidence that bacterial ribosomes are attached to membrane solely by their secreted chain . The present results further show that after loss of the extracellular segment of the chain its attachment persists, at 37 degrees as well as 0 degrees C . These findings suggest that the chain does not slip through a passive membrane but is actively held within a channel. J Virol, 1978 Dec, 28(3), 697 - 709 Relationship of Bacillus subtilis DNA polymerase III to bacteriophage PBS2-induced DNA polymerase and to the replication of uracil-containing DNA; Hitzeman RA et al.; In vivo studies of PBS2 phage replication in a temperature-sensitive Bacillus subtilis DNA polymerase III (Pol III) mutant and a temperature-resistant revertant of this mutant have suggested the possible involvement of Pol III in PBS2 DNA synthesis . Previous results with 6-(p-hydroxyphenylazo)-uracil (HPUra), a specific inhibitor of Pol III and DNA replication in uninfected cells, suggest that Pol III is not involved in phage DNA replication, due to its resistance to this drug . Experiments were designed to examine possible explanations for this apparent contradiction . First, assays of the host Pol III and the phage-induced DNA polymerase activities in extracts indicated that a labile Pol III did not result in a labile phage-induced enzyme, suggesting that this new polymerase is not a modified HPUra-resistant form of Pol III . Indeed the purified phage-induced enzyme was resistant to the active, reduced form of HPUra under all assay conditions tested . Since in vitro Pol III was capable of replicating the uracil-containing DNA found in this phage, the sensitivity of the purified enzyme to reduced HPUra was examined using phage DNA as template-primer and dUTP as substrate; these new substrates did not affect the sensitivity of the host enzyme to the drug. J Bacteriol, 1978 Dec, 136(3), 886 - 99 Cell wall assembly in Bacillus subtilis: location of wall material incorporated during pulsed release of phosphate limitation, its accessibility to bacteriophages and concanavalin A, and its susceptibility to turnover; Anderson AJ et al.; Addition of a pulse of phosphate to a phosphate-limited chemostat culture of Bacillus subtilis W23 led to the synthesis of teichoic acid and the consequent development by the bacteria of the ability to bind phage SP50 . In cultures growing at different rates, phage-binding properties became maximal approximately one generation time after addition of the pulse . Removal of the incorporated teichoic acid by turnover also reached its maximum rate after a similar interval . After pulsed release of phosphate limitation in B . subtilis NCTC 3610, the alpha-glucosyl residues of the incorporated teichoic acid, detected by their interaction with concanavalin A, became maximally exposed at the same time that phage binding was maximum . At that time the bacteria bound phage all over the cylindrical part of the surface and at about one-third of the polar caps . That fraction of the receptor material that is exposed soon after its incorporation was distributed along the cylindrical length of most of the bacteria, but few phages bound to the polar caps, except in the case of short bacteria; these bound phages in a markedly asymmetric manner at one pole and along their length . The significance of these results is discussed in relation to the mode of assembly of the cell wall. J Bacteriol, 1978 Dec, 136(3), 883 - 93 Suppression of temperature-sensitive sporulation of a Bacillus subtilis elongation factor G mutant by RNA polymerase mutations; Hirochika H et al.; A class of rifampin-resistant (rfm) mutations of Bacillus subtilis suppresses the temperature-sensitive sporulation of a fusidic acid-resistant mutant . FUS426, which has an altered elongation factor G . The rfm mutation suppressed only the sporulation defect caused by the elongation factor G mutation, but could not suppress other types of induced sporulation defects . Genetic and biochemical analyses showed that the sporulation suppression by the rfm mutation was caused by a single mutation in RNA polymerase . After the early sporulation phase, the apparent rate of RNA synthesis of FUS426, measured by {3H}uracil or {3H}uridine incorporation into RNA, became lower than that of the wild-type strain, and this decrease was reversed by the rfm mutation . However, when the total rate of RNA synthesis of FUS426 was calculated by measuring the specific activity of {3H}UTP and {3H}CTP, it was higher than that of the rfm mutant, RIF122FUS426 . The possible mechanism of the functional interaction between elongation factor G and RNA polymerase during sporulation is discussed. J Bacteriol, 1978 Dec, 136(3), 854 - 66 Alterations in Bacillus subtilis transforming DNA induced by beta-propiolactone and 1,3-propane sultone, two mutagenic and carcinogenic alkylating agents; Kubinski ZO et al.; Transforming DNA was exposed to either beta-propiolactone or 1,3-propane sultone and then used for transformation of competent bacteria to nutritional independence from tyrosine and tryptophan (linked markers) and leucine (an unlinked marker) . The ability to transform was progressively lost by the DNA during incubation with either of these two chemicals . For all three markers the inactivation curve was biphasic, with a short period of rapid inactivation followed by one characterized by a much slower rate . The overall rate of inactivation was different for all three markers and presumably was related to the size of the marker . The decrease in the transforming activity was in part due to the slower rate of penetration of alkylated DNA through the cellular membrane and its inability to enter the recipient bacteria . This decrease in the rate of cellular uptake, even for DNA eventually destined to enter the cell, began almost immediately after its exposure to the chemical and ended up with an almost complete lack of recognition of the heavily alkylated DNA by the specific surface receptors of competent cells . Such DNA attached to sites on the surface of competent bacteria which were different from receptors specific for the untreated nucleic acid . This attachment was not followed by uptake of the altered DNA . Presence of albumin during the incubation with a carcinogen further increased the degree of inactivation, indicating that the artificial nucleoproteins produced under such conditions were less efficient in the transformation assay than was the naked DNA . Cotransfomration of close markers progressively decreased, beginning immediately after the start of incubation of DNA with the chemicals . Extensively alkylated DNA fractionated by sedimentation through sucrose density gradients showed a peculiar distribution of cotransforming activity for such markers; namely, molecules larger than the bulk of DNA ("megamolecules") showed less ability to transform the second marker than did some of the apparently smaller molecules which sedimented more slowly through the gradient . An increase in cotransformation of distant markers was evident in DNA molecules after a short exposure to an alkylating agent, but cotransformation of such markers was absent in DNA treated for longer periods . The observed changes in the transforming and cotransforming activities of the alkylated DNA can be explained by what is known about the physicochemistry of such DNA and in particular about the propensity of the alkylated and broken molecules to form complexes with themselves and with other macromolecules. J Bacteriol, 1978 Dec, 136(3), 1205 - 7 Differential effect of hydroxyurea on the replication of plasmid and chromosomal DNA in Bacillus subtilis; Shivakumar AG et al.; The replication in Bacillus subtilis of the staphylococcal R plasmids pE194, pBD15, pUB110, pSA0501, and pSA2100 has been studied in the presence of hydroxyurea . In all cases, an enrichment for covalently closed circular DNA compared with chromosomal DNA was observed . In this respect, hydroxyurea mimics the effect previously observed with pUB110, using strains carrying the conditional mutation dnaA13 . This mutation has been reported to affect ribonucleotide reductase (G . W . Bazill and D . Karamata, Mol . Gen . Genet . 117:19-29, 1972) . An explanation for these effects is offered, together with some supporting evidence. Mol Gen Genet, 1978 Nov 29, 167(2), 157 - 64 SPP1 DNA replicative forms: growth of phage SPP1 in Bacillus subtilis mutants temperature-sensitive in DNA synthesis; Mastromei G et al.; The development of bacteriophages SPP1 and phi 29 has been studied in several B . sutilis mutants defective in host DNA replication, under non permissive conditions . Several gene products, involved in the synthesis of host DNA, are required for phi 29 replication, while SPP1 seems to require only the host DNA polymerase III . In addition both phages are unable to grow in a dna A mutant (ribonucleotide reductase) . Taking advantage of the fact that SPP1 DNA is actively replicated in several dna mutants at non-permissive temperature, we have studied the structure of the replicative intermediates of this phage in the absence of interfering host DNA synthesis . Fast sedimenting forms of SPP1 DNA can be isolated from phage infected cells and evidence of covalently joined concatemers has been obtained, suggesting the presence of terminally repeated sequences. Biochim Biophys Acta, 1978 Nov 21, 521(1), 16 - 26 Induction of sporulation by inhibitory purines and related compounds; Heinze JE et al.; Sporulation of Bacillus subtilis can be induced, in the presence of excess ammonia, glucose and phosphate, by many purine derivatives under conditions of partial growth inhibition . Some of the compounds are known inhibitors of purine nucleotide synthesis . For most compounds the effect is counteracted by adenine and guanine . Partial growth inhibition by amethopterin (methotrexate) causes sporulation in the absence of purines but not in their presence . Unable to induce sporulation at any concentration are inhibitors of DNA, RNA, and protein synthesis as well as base or amino acid analogs that are incorporated into these polymers. Biochim Biophys Acta, 1978 Nov 15, 544(1), 1 - 7 Regulation of lactate dehydrogenase synthesis in Bacillus subtilis; Yashphe J et al.; The regulation of lactate dehydrogenase in Bacillus subtilis was determined under a variety of growth conditions and in mutants blocked in the citric acid cycle . The synthesis of lactate dehydrogenase increased sharply concomitantly upon the exhaustion of glucose from the medium and the onset of the stationary phase . The synthesis of lactate dehydrogenase may be under catabolite repression control . Studies with mutants blocked in the citric acid cycle showed that lactate dehydrogenase is regulated independently of either the oxidative or reductase branches of the cycle . Certain citric acid cycle mutants, e.g., aconitase or succinate dehydrogenase, exhibited very low levels of lactate dehydrogenase while others, e.g., malate dehydrogenase or isocitrate dehydrogenase, showed normal levels . A stage O sporulation mutant expressed levels of lactate dehydrogenase more than one thousand-fold higher than the low group of citric acid cycle mutants . The induction of lactate dehydrogenase was shown to be independent of the accumulation of its substrate, pyruvate. Mol Gen Genet, 1978 Nov 9, 166(3), 287 - 90 Genetic exchange in Bacillus subtilis in soil; Graham JB et al.; Genetically labelled strains of Bacillus subtilis have been shown to exchange blocks of linked genes while growing together in soil . After eight days of incubation, 79% of unselected colony-forming units exhibited a phenotype containing markers from both parents; the parental strains were not detected after the first day of incubation . High frequencies of transformation were also obtained by adding genetically labelled deoxyribonucleic acid to single-strain soil cultures . Observed linkage of genetic markers was greater in soil transformation than in standard laboratory procedures . The results indicate that transformation may play an important role in the adaptation of the Bacilli to their natural habitat. Ann Microbiol (Paris), 1978 Nov-Dec, 129 B(4), 537 - 49 Pleiotropic control mutations affecting the sporulation of Bacillus subtilis; Balassa G et al.; Mutations affecting quantitatively the production of the sporulation-associated extracellular alkaline protease were isolated and characterized . They fall into at least five genes, three of which, ScoA, B and C, were mapped in the argC-metC region . The pleiotropic effects of these mutations concern several or all of the following: rate and timing of protease production, synthesis of alkaline phosphatase, time-course of spore formation . Electron microscopic evidence indicates delayed switch from one morphological stage to another . The nature of the Sco mutations and the genetic regulation of sporulation are discussed. Prikl Biokhim Mikrobiol, 1978 Nov-Dec, 14(6), 890 - 7 {Preparation of extracellular ribonuclease from Bacillus subtilis}; Bashkis EV et al.; The paper describes a method for isolating alkaline ribonuclease from the culture liquid Bacillus subtilis KP 349 which involved: submerged cultivation of the producer on complex and synthetic nutrient media with optimized RNase activity, acid treatment of the total culture liquid, and filtration through perlite . Further treatment may include either spray drying of the culture liquid filtrate or its concentration in a vacuum evaporator, dialysis of the concentrate against distilled water, and dialyzate lyophilization . As a result, commercial RNase preparations with activities of 30--60 thous . and 160--300 thous . units/g, respectively, were obtained . The enzyme purification was carried out by chromatography and rechromatography on phosphocellulose columns . The resultant RNase of Bac . subtilis had a specific activity of 41--44 thous . units/mg protein, contained no nonspecific phosphodiesterase, DNase, acid or alkaline phosphomonoesterases. Can J Microbiol, 1978 Nov, 24(11), 1431 - 3 Survival of microbial films in the microwave oven; Page WJ et al.; Air-dried films of Escherichia coli, Saccharomyces cerevisiae, and Bacillus subtilis spores on membrane filters, exposed to 10 min full power (650 W, 2450 MHz) irradiation in a Toshiba model ER776BT microwave oven, showed a 5-, 2-, and 0-log reduction of viable organisms respectively . Suspensions of cells or spores in phosphate buffer treated under similar conditions showed 8 logs of killing within 30 s (S . cerevisiae), 45 s(E . coli), and by 10 min (B . subtilis spores) of exposure. Genetika, 1978 Nov, 14(11), 2046 - 8 {UV-light induction of "true" revertants to adenine independence in Bacillus subtilis cells}; Lotareva OV; The UV-irradiation of Bacillus subtilis Mu5u8u16 (met5 leu8 purA) induces with relatively high frequency the revertants to adenine independence (Ade+) which form the rapidly growing morphologically uniform colonies on the solid selective medium . The genetic analysis of a portion of UV-induced Ade+ revertants (crosses in transformation system) has cleared up that their DNA does not contain the original mutation ade16 . This means that they arise as the result of "true" reversions . This reversion in purA gene can serve as a good model for the study of UV-induced mutagenesis in a proximal structural locus of Bac . subtilis chromosome. Genetika, 1978 Nov, 14(11), 1900 - 7 {Effect of Bacillus subtilis A-50 "streptomycin" mutations on the formation of molecular forms of subtilisin, an extracellular alkaline proteinase}; Ermakova LM et al.; The patterns of subtilisin molecular forms of streptomycin-resistant (Strr) and streptomycin-dependent (Strd) mutants of Bacillus subtilis A-50, as well as the revertants of Strd to streptomycin-independence (Str1) were studied . Strr mutants had different quantitative pattern of the same subtilisin molecular forms as compared with the initial strain A-50 (the forms with Rf 0.08, 0.16 and 0.3) . In comparison with the initial strain A-50, Strd mutants and Str1 revertants revealed three additional forms of the active enzyme with Rf 0.02, 0.5 and 0.7 and the molecular weights less than 35,000, 28,000 and 20,000 respectively . It was suggested that the rate and character of the enzyme secretion of the degree of its post-translational modifications might result in the different pattern of subtilisin molecular forms produced by these streptomycin mutants. Biull Eksp Biol Med, 1978 Nov, 86(11), 556 - 7 {Effect of dimethyl sulfoxide on transformation of B . subtilis in vitro}; Orlova EB et al.; Bacillus subtilis transformation was conducted in the presence of dimethylsulfoxide and polyethyleneglycol . B . subtilis transformation was most frequent under the effect of 0.1% dimethylsulfoxide and was not altered significantly by polyethyleneglycol . As suggested, an increase of the transformation frequency was associated with the change of the membrane permeability under the influence of dimethylsulfoxide of with the altered activity of the membrane DNase. J Bacteriol, 1978 Nov, 136(2), 818 - 21 Tunicamycin-resistant mutants and chromosomal locations of mutational sites in Bacillus subtilis; Nomura S et al.; The types of tunicamycin-resistant mutants of Bacillus subtilis were analyzed, and their mutational sites on the chromosome were mapped . A type 1 mutation that simultaneously expressed hyperproductivity of extracellular alpha-amylase was located close to amy E . Type 2 mutations were near aroI. J Bacteriol, 1978 Nov, 136(2), 799 - 802 Chromosome-membrane association in Bacillus subtilis . IV . Further purification of DNA-membrane complex by using a combination of centrifugation and electrophoresis; Toyoda H et al.; We have developed a simple procedure to purify a DNA-membrane complex from Bacillus subtilis by using a combination of centrifugation and electrophoresis . Several unique proteins were detected in the purified complex. J Bacteriol, 1978 Nov, 136(2), 484 - 90 Inhibition of Bacillus subtilis spore germination by various hydrophobic compounds: demonstration of hydrophobic character of the L-alanine receptor site; Yasuda-Yasaki Y et al.; L-Alanine-initiated germination of Bacillus subtilis spores was inhibited by various kinds of hydrophobic compounds . Good correlation of inhibitory effect with hydrophobicity of the compound was demonstrated by using regression analysis in which the hydrophobic character was expressed by the partition coefficient in an octyl alcohol-water system . The correlation coefficient for 20 alcohols was 0.959, and that for 19 miscellaneous compounds was 0.906 . Regression lines of the alcohols and other hydrophobic compounds were almost identical, showing that hydrophobic interaction played an important role in inhibition . Diphenylamine was one of the most effective inhibitors examined . n-Octyl, n-nonyl, and n-decyl alcohols were the most effective alcohols . The mode of inhibition by diphenylamine and n-octyl alcohol was a "mixed type" (competitive plus noncompetitive type) with respect to L-alanine; that by D-alanine was competitive inhibition . Sites for diphenylamine, n-octyl alcohol, and D-alanine may have overlapped . Inhibition was reversible by washing; heat resistance, stainability, and germination rate of the washed spores remained unaltered . Thus, we confirmed that the inhibition may occur before the initial trigger reaction of germination and that it may be due to the interaction between a hydrophobic compound and a hydrophobic region closely associated with the L-alanine receptor site on the spore. Cancer Res, 1978 Nov, 38(11 Pt 1), 3918 - 21 Carcinogenicity of triethanolamine in mice and its mutagenicity after reaction with sodium nitrite in bacteria; Hoshino H et al.; Mice fed a diet containing 0.3 or 0.03% triethanolamine developed malignant tumors . Females showed a high incidence of tumors in lymphoid tissues, while this type was absent in males . Tumors in other tissues were produced at a considerable rate in both sexes, but no hepatoma was found . Triethanolamine was not mutagenic to Bacillus subtilis by itself, but it became mutagenic after reacting with sodium nitrite under acidic conditions or when the mixture was heated . Although N-nitrosodiethanolamine, a known carcinogen and mutagen, was detected in the reaction mixture by thin-layer chromatography, it may not be the main mutagenic product, because the product was a stable and direct mutagen and its mutagenic activity was destroyed by liver enzymes, unlike N-nitrosodiethanolamine . The lethal and mutagenic DNA damages produced by this unidentified product were susceptible to some extent to the repair functions of the bacteria. J Bacteriol, 1978 Nov, 136(2), 625 - 30 Benzeneboronic acid selectively inhibits sporulation of Bacillis subtilis; Davis-Mancini K et al.; m-Aminobenzeneboronic acid at levels of 0.2 mM in nutrient broth medium selectively inhibited sporulation without appreciably altering vegetative growth . Significant inhibitory effects were seen even when it was added as late as 6 h after the end of logarithmic growth . The pH changes associated with growth and sporulation of Bacillus subtilis in nutrient broth were not significantly altered by the inhibitor . When it was present in cultures of actively growing cells, its inhibitory effect could not be reversed by simple dilution . The compound caused extensive clumping, of cells, which appeared not to be related to the ability of boronates to esterify to diols. Mol Gen Genet, 1978 Oct 30, 166(2), 119 - 26 An unstable donor-recipient DNA complex in transformation of Bacillus subtilis; Popowski J et al.; In re-extracted DNA obtained shortly after uptake of transforming DNA by Bacillus subtilis, increased amounts of donor DNA radioactivity banding at the position of donor-recipient DNA complex (DRC) are observed in CsCl gradients, if the cells are irradiated with high doses of UV prior to reextraction of the DNA . Qualitatively, the same phenomenon is observed if lysates of transforming cells are irradiated . UV-irradiation of lysates of competent cells to which single-stranded DNA is added after lysis, does not result in linkage of this DNA to the chromosomal DNA . Two observations argue in favour of the formation of a specific labile complex between donor and resident DNA during transformation . Firstly, heterologous donor DNA from Escherichia coli, although being processed to single-stranded DNA in competent B . subtilis, does not seem to be linked to the recipient chromosome upon UV-irradiation, and secondly, the labile complex of donor and recipient DNA can be stabilized by means of treatment of the lysates of transforming cells with 4, 5(1), 8-trimethylpsoralen in conjuction with long-wave ultra violet light irradiation . This indicates that basepairing is involved in the formation of the complex . On the basis of these results we assume that the unstable complex of donor and recipient DNA is an early intermediate in genetic recombination during transformation. Eur J Biochem, 1978 Oct 16, 90(3), 581 - 3 Restriction and modification in Bacillus subtilis . Localization of the methylated nucleotide in the BsuRI recognition sequence; Gunthert U et al.; Calf thymus DNA was methylated in vitro with cell extracts of Bacillus subtilis OG3R (r+m+) and S-adenosyl{Me-3H}methionine . After depurination of the {3H}methylated DNA, the analysis of the pyrimidine dinucleotides revealed the following positions of the methylated nucleosides (indicated by an asterisk) within the BsuRI recognition sequence: 5' dG--dG--dC--dC dC--dC--dG--dG 5'. J Biol Chem, 1978 Oct 10, 253(19), 6694 - 701 Spore coat protein of Bacillus subtilis . Structure and precursor synthesis; Munoz L et al.; The coat protein of Bacillus subtilis spores comprises about 10% of the total dry weight