Journal of Bacteriology, April 2002, p . 2065-2071, Vol . 184, No . 8

Thanks, Charley

Michael D . Manson^*

Department of Biology, Texas A&M University, College Station, Texas 77843-3258

 
How many molecular biologists have orchids named for them? How many have influenced three generations of science as Charley has? How often, in any discipline, can you mention a first name and have everyone know whom you mean? And how many names are synonymous with a molecule? Charley equals tryptophan .

Eric Selker remembers his early days as a graduate student . "I told Charley I was interested in eukaryotic gene regulation . Charley said that was fine, as long as my project had to do with trp . I read whatever I could on trp regulation in yeast, peas, Neurospora, etc." Eric decided on Neurospora . One wonders what would have happened had Eric chosen peas .

Charley was not a late bloomer . Paul Berg recalls, "Charley had already acquired giant status when I was a postdoc . His lecture at a symposium in Detroit in 1955 or 1956 (Charley was then at Case Western) was the clearest evidence I knew that supported the direct gene-enzyme coding relationship implied by Beadle and Tatum's one gene-one enzyme model proposed 10 years earlier . It was especially gratifying when he stopped at Washington University and stayed at my house on the way to be interviewed for Ed Tatum's position in Stanford's Biology Department . He knew that Arthur Kornberg and the entire Department of Microbiology had agreed to create a new Department of Biochemistry at Stanford . Persuaded by that, Charley accepted Stanford's offer in spite of the less-than-adequate lab facilities that would be available to him at the time."

Those antiquated facilities in the old Jordan Hall did not quench the lab's esprit . Milton Taylor, one of Charley's first graduate students at Stanford, recalls "I arrived in 1961 . I was in the oldest Yanofsky lab, in the basement . It was very crowded . Journal club was the highlight of the week . We met in the seminar room . After the session we went to Charley's house for coffee, cake, and ice cream, provided by Carol, who was a gracious hostess . Wives were included . We played croquet after dessert . These "scientific-social' events cemented the lab . We functioned as a team . When I set up my lab, I organized a journal club along the same lines, substituting bowling for croquet."

Charley has a knack for making your life easier if you let him . I was reminded of this gift after I offered to write this piece . Without knowing what I was planning, Charley sent me a reprint of his scientific autobiography, "Advancing Our Knowledge in Biochemistry, Genetics, and Microbiology through Studies on Tryptophan Metabolism" (8) . The title is typically Charley—to the point rather than cute or zippy . The text makes fascinating reading for anyone interested in learning how Charley views the most significant and influential events in his professional life . The major discoveries are all there: colinearity of gene and protein, translational suppression, transcription attenuation, the basis of trp operator DNA/Trp repressor interaction, and control of conidiation in Neurospora . (1-7, 9) . This happy coincidence leaves me free to elaborate on what Charley would not—the profound and enduring effect he has had on students, colleagues, collaborators, and friends—admirers all .

Charley is aware of this legacy . The abstract of his review states, "I was fortunate to practice science during the last half of the previous century, when many basic biological and biochemical concepts could be experimentally addressed for the first time...Throughout my career I enjoyed the excitement of solving basic scientific problems . Most rewarding, however, was the feeling that I was helping young scientists experience the pleasure of performing creative research." What follows are the recollections of those scientists, all young when we first met Charley, some of us not so young anymore .

Charley played to win . These contests were legendary even beyond the Yanofsky lab . Fred Alt, a student with Bob Schimke at the time, was good at table tennis . In a dream, Fred found himself at the Yanofsky's for the first time . After the presentations, everyone migrated to the kitchen for cake and cookies, as Fred had heard . Charley came up to Fred with a welcoming smile, put his hand on Fred's shoulder, and gestured toward the den . "A little Ping-Pong, Fred?"

I also remember a basketball game between the Yanofsky and Schimke labs . We graduate students and postdocs confined ourselves to outside shots . It was dangerous inside, where Bob and Charley jockeyed for rebounding position . This no-holds-barred approach to hoops extended to pickup games in the Yanofsky driveway involving Charley, his three sons, and their friends . Matt Bonner recalls, "I spent many summers at their house playing basketball and baseball (well, let's face it, every sport invented) and swimming . I remember one basketball game when Charley was determined not to let the kids win . He twisted his ankle making a run at the basket over any kid in the way . However, it was all in good fun, and never personal."

Sunday-morning tennis consisted of matches among Charley, Paul Berg, another Stanford faculty member or two, and sometimes a decidedly nervous postdoc or graduate student . Charley mentions these contests in his Annual Review of Biochemistry memoirs (8), and Paul Berg also looks back on them fondly . "Having settled on the Stanford campus just down the street from Charley's house, our two families became one, and we soon discovered that our passions for science were matched by our intensely competitive drives on the tennis court . What better way to spend Sunday mornings than facing each other across the net, determined to do each other in, and then ending up with an in-depth retelling of what went on in our respective labs during the previous week . Ideas flowed freely, often culminating in experiments to test them . It was in that setting that our one collaboration—proving that missense suppression results from mutant tRNAs—originated . It was then that I first began to appreciate and use the power of microbial genetics."

When I worked with Charley (1969 to1975) it was not obvious that he competed against anyone but himself in science . He seemed unconcerned about getting scooped . I once asked Charley where he was going on sabbatical . He replied that he would stay in his lab at Stanford because that was where the most exciting stuff was going on . This attitude was reassuring for graduate students and postdocs . It provided security and time to develop at our pace in the best place in the world . It also kept us in science . We were shielded from the unpleasant aspects of the business . If Charley ever sweated a grant deadline, it did not show .

Cathy Squires, who came with her husband Craig as a postdoc in the early 70s, remembers, "Charley gets more high-quality work from anyone than any scientist I have met . His weekly meetings with each person were the epitome of encouragement . You went away filled with ideas to pursue and excitement about what you were doing . He never made you feel you had let him or the project down—you just had to try more things! Charley clearly didn't concern himself with what you did from minute to minute, but he expected some progress each week, and that's what he got." Well, sometimes .

Cathy Squires learned that you reject Charley's advice at your peril . "When I started, I was sure I knew far more than Charley about the in vitro system I was setting up . So, I listened to his suggestions politely and ignored them! After six months, I realized that some of his suggestions would have led to far quicker success if I had followed them . I mended my ways."

Sometimes what Charley noticed were details that were not quite in place . My own vivid memory comes from a conversation we had about the third manuscript in my dissertation . We submitted the other two to the Journal of Bacteriology, and Charley asked where I wanted to send the third one . It was obvious that he did not intend to be an author . I asked what was wrong . He could not put his finger on it, but somehow my data did not convince him . (Subsequent work by Irv Crawford and Ray Mosteller showed that Charley's instinct was correct.) Although he did not speak disparagingly, I was peeved . I asked why he thought I would submit a paper that was not good enough for him . The only answer I remember was a wry smile . I vowed then and there never to submit a paper that did not deserve to have Charley Yanofsky on it . I have not lived up to that standard, but it remains a fine one .

Eric Selker expresses his wonder at Charley's efficiency . "Back in the days before e-mail, prominent scientists like Charley received a great deal of mail . Every morning, Charley walked through the grad lab (my guess is that he designed the lab with this path in mind) with a huge load of mail, weighing perhaps 10 pounds . He would then start answering the mail, scanning the journals, etc., which would take most of us the whole day, but within an hour he was done."

Many of us benefited from Charley's promptness in correcting manuscripts . Matt Sachs notes, "One thing that comes to mind immediately is the awesome way that Charley handles drafts of manuscripts (overnight when I was in the lab) and the thorough way he criticizes them." Eric Selker adds, "The most remarkable example that I recall concerns a set of papers on attenuation that took up half of one issue of the Journal of Molecular Biology . Charley edited all the manuscripts in one evening, despite his having been somewhat under the weather that day." Jack Rose shares similar thoughts . "When students or postdocs wrote papers, Charley would have extensive comments and revisions back the next day . He taught science writing just as vigorously as he taught science."

Vigorous is right . I decided to inject literary flair into the first manuscript I submitted to Charley . The next day it was on my desk with a terse note . "Nobody wants to read all this garbage . Try again." My prose may still be "wordy" and "long-winded," but not on purpose .

My 5-year postdoc was followed by 4 years as a research scientist in Germany . I finally realized that it was "now or never" if I wanted a faculty position in the United States . I wrote Charley in some trepidation to ask if he could provide a positive recommendation . His reply began, "The letters I write are fair." I did not need to read further . If I deserved a job, Charley's letter would help me, warts and all, to get it .

Eric Selker wrote, "People quickly recognize that Charley is knowledgeable, imaginative, and kind . It soon also becomes obvious that he is extremely gifted in the lab, that his instincts are fantastic, and that his efficiency is unparalleled . Every motion and every word count . Charley was a father figure; the German ‘Doktorvater' seems to fit perfectly . A father's most important function is to set a good example . Charley was, and continues to be, a great model . I have found it impossible to come close to his example in most areas, but it continues to provide guidance."

Terry Platt summarizes his feelings thus . "I fondly remember the golden years of the mid-1970s as a postdoc in Charley's lab (1972 to 1975) . The literature documents the elucidation of the mechanism of attenuation . I was also fascinated by the way Charley ran a successful lab and what I could learn to help me later as a faculty member . His group felt like a large family, full of mutual affection and sibling rivalry . Charley, as patriarch, spurred us on with curiosity and enthusiasm, rarely expressing impatience . He embraced all genuine effort, an approach that I try to emulate in my own laboratory . There was also a remarkable convergence of multiple lines of inquiry to understand attenuation . The philosophy was that a laboratory should work on an overall problem—tryptophan metabolism in Charley's case—rather than addressing disparate questions . Everyone could talk with everyone and share a common interest despite the apparent unrelatedness of specific projects . When those projects pointed in the same direction, the realization we were part of a team added energy to the thrill of discovery . Finally, over the past three decades Charley has continued to follow and encourage my efforts to carve a research niche of my own . I will always cherish having Charley as mentor and friend over this time."

Charley was always excited when he learned new things from students . Paul Babitzke, a postdoc from 1991 to 1994, looks back on such a moment . "I was studying the trp/TRAP system in Bacillus subtilis . I was testing the attenuation model proposed by Charley and Mitzi Kuroda . Charley and I designed an in vitro filter-binding experiment to determine if RNA secondary structure occluded TRAP from one of two 10-nucleotide sites in the leader . My data made no sense .

"I had to give group meeting the next morning, so I was trying to figure out how to explain the data . I examined the trp leader sequence, but I was constantly interrupted by labmates who wanted to chat, Charley included . As a result, I kept losing my place . Finally, I realized I kept getting lost because there were 11 GAG and UAG repeats within the leader sequence, separated by two or three nucleotides . These repeats overlapped the TRAP-binding sequences that Mitzi and Charley had identified . The more repeats, the tighter the binding . The trpG transcript had nine GAG repeats overlapping the Shine-Dalgarno sequence . I was convinced I had the answer."

"I prepared an overhead and confidently presented it at the group meeting . I was reading off ‘GAG, space, space, GAG, space, space, UAG, etc.' when Charley suddenly said ‘That's enough, sit down!' I was stunned, my confidence visibly shaken . On the way back to the lab, someone joked that Charley had invoked the ‘GAG' rule . Ten minutes later Charley burst into my office . ‘Do you think this is right? Here's what we need to do.' Charley had not interrupted because he was annoyed . He wanted to get back to his office to check the leader sequences . This day was, perhaps, the most exciting in my career . I, of all people, had taught Charley something."

Charley expressed his view on mentoring to me in a 1998 letter . Some of it was personal and brought a smile . Charley did understand me! His more general comments illustrate what Charley means to those who work with him . "I didn't know what to make of your letter—are you retiring? Kidding aside, from my perspective it is nice to know that at least one of your students appreciated you! Handling students is much like dealing with your own children . Are you more helpful when you are tolerant and patient, or when you are critical—who knows? Each of us is different . It is probably a mistake to assume that there is only one correct approach . In your case...You also made a wonderful choice in picking Lily [my wife, Lily Bartoszek, assistant to Journal of Bacteriology editors Susan Golden and Ry Young] . She is delightful and must be a great companion."

Being married to Carol and, after Carol succumbed to cancer in 1990, Edna, widow of Charley's close friend and scientific colleague Irv Crawford, has given Charley rich experience with wonderful companions . He has also been blessed with loyal and talented assistants: Miriam Bonner and Ginny Vania as technicians and Susan Lacoste as secretary . Aside from their awesome professional skills, they always listened to students' problems and gave sensible advice, whether about science, Charley, or the travails of life in general . The women in his life helped keep Charley human . We loved them for it, and for themselves .

Roberto Kolter, postdoc in the early 1980s, also retains memories of Friday meetings . "I was always excited to talk to Charley on Friday . A few times I had results to discuss, but it did not matter . The beauty of going into Charley's office was that, for those minutes, I had the undivided attention of one of the world's brightest geneticists/biochemists . Brainstorming with Charley was the best intellectual experience of my Stanford years . We would dream up genetic selections, new techniques, and new ways of looking at results . The vast majority of these ideas never were put to the test . However, the excitement we felt was, for me at least, reward enough."

Jack Rose also focused on the plus side . "Charley made himself available in daily visits to the bench and scheduled weekly office visits for the standard inquiry, ‘What's new?' I cannot imagine a better atmosphere for a graduate student." Terry Platt adds, "Charley taught me to hold weekly conferences with students and postdocs, always on Friday . No matter how frustrating the week had been, Charley generated a set of incisive questions and encouraging possibilities to reenergize us for the next week . Some people came in over the weekend to start the next round of experimental troubleshooting to generate preliminary answers by Monday."

Allan Campbell noted Charley's preference for no frills . "This is kind of a nonresponse . One time, long ago, Charley introduced me for a seminar . He remarked that he had considered including some personal anecdotes but had then reflected that some day I might be introducing him . Suffice it to say I have a tremendous admiration and affection for Charley." Allan's story reminds me of when I hosted Charley at Texas A&M a few years ago . Since it was a sponsored guest lectureship, complete with a commemorative plaque, and because university reporters and photographers were on hand, I had prepared a rather long list of Charley's awards, honors, and accomplishments . I had barely launched into my introduction when Charley, in the first row, said in a stage whisper "If you keep on like this, I won't have time to talk." That stopped me cold . I tried to cover my chagrin with a segue to, "And so, without further ado, here's Charley!" You will not find a list of awards and honors here . They are a matter of public record .

Bob Fisher, who was with Charley from 1981 to 1983, provides the perspective of someone who made the transition from postdoc to colleague . "I stayed at Stanford since that time, working with Sharon Long, so I have continued to see Charley for the last 20 years . My strongest impression is the model that he sets to this very day . He is still in the lab, doing experiments . He calls me up occasionally to ask me to look at a column he has poured or to review the use of the French press . I have been tickled by this classic example of the master asking the student for guidance . The man loves science and the discovery of new information . He wants to be involved in the process, hands-on . He is also extremely generous with equipment, materials, and space, and our lab has benefited on numerous occasions . He's the best."

This is an opinion shared by Jim Spudich, who got his Ph.D . with Arthur Kornberg and then did a 1-year postdoc with Charley in 1968 . Jim subsequently returned to Stanford as a faculty member in Biochemistry . "Charley is one of my few heroes, not only as a scientist, but as a special human being . I feel deeply honored to have worked with him and to have known him all of these years . He has been a role model for me . They just don't come any better than Charley."

Charley's visits to the bench could come too often when you were goofing off . Ford Doolittle confesses after 30+ years . "I was not one of Charley's hardest working graduate students . When he popped into the lab, I often did not have much to say in response to his interrogatory ‘So?' After several embarrassments when I was caught snoozing with nothing new to report, I developed a foolproof stratagem . I kept a rack of tubes containing the colorful indole assay in the drawer of my desk . When Charley exited his office, I bustled off to read them in the Klett, too busy for idle chatter . If the tube contents faded or evaporated, I'd replace them . Charley never said whether he found this suspicious . Now, when I ask my students about their progress I hear Charley's ‘So?' and wonder if they have similar tricks."

Charley did not trigger these visions and subterfuges by what he said or did . They occurred because Charley was Charley—a model for excellence who could cope with anything . That tended to get you down when you could not finish your small piece of the big Yanofsky pie .

That was not a permanent solution, however, so Manny finally went to Charley "to discuss my future (or lack thereof) in science . He expressed confidence in my abilities, encouraged me to continue what I was trying to do, and supported my staying for another year beyond the two for which I had a fellowship . Except for his understanding, I might today be playing saxophone (Fig . 2, right) on some street corner in San Francisco rather than doing the scientific research and teaching I so enjoy!"

 
Charley fostered the careers of many scientists . Roberto Kolter came to "iron out the wrinkles of attenuation in gram-negative bacteria," but his research interests wandered . "Charley never showed concern when I got involved in projects throughout the campus . I appreciated the freedom to grow as a scientist and an individual . It was vital to my development . I arrived enamored with the miracles his lab had performed with bacteria . I left confident that I could explore whatever question I dreamt of in bacteria because of the wonders that could be done with them . Charley's love for microorganisms proved contagious, and I left more passionate about the microbial world . My work over the 20 years since I left Charley's lab has nothing to do with tryptophan, but it bears the imprint of the intellectual impression Charley left in me."

Naomi Franklin has known Charley longer than anyone . "Charley was already a driving wheel in the Bonner lab at Yale when I started graduate studies in 1950 . The scientific ferment was just what I hoped to find . The language seemed Chinese those first 6 months, but with the help of Charley and others, I began to catch the lingo . Enhanced by camaraderie among a wild assortment of personalities, the pleasure of science I experienced then has lasted a rich lifetime .

"My interest in phage proved valuable when I joined Charley's lab at Stanford in 1963 . Phage 's cousin 80 resides in the Escherichia coli chromosome next to the trp operon . Deletions fusing trp to a /80 hybrid prophage integrated at this site, providing direct proof for Allan Campbell's idea that the prophage inserts linearly, with permuted gene order, into the E . coli chromosome .

"The dependence of phage on N protein involves overcoming intrinsic transcription terminators within the N operon . Polarity induced by nonsense mutations in the trp genes also proved to be due to transcription termination, reversible in vivo by N protein . Charley's lab discovered a related phenomenon, attenuation in the trp operon, about this time (in 1973) . The notion that mRNA structure can regulate gene expression was new in the early seventies and undoubtedly influenced my subsequent thinking about the nature of N antitermination .

"Very early in this epic, Charley set me upon a course of research autonomy by endorsing my application for an independent grant . NSF proved willing and supported the work over the next thirty-some years . Charley's initiative set things in motion . I also remain grateful for 16 years of haven in Charley's stimulating circle: great science, lively colleagues, and the friendship of Carol and the Yanofsky boys . Miriam Bonner also found a productive haven with Charley, enhancing our family circle . The big professional lesson that I took away from those years was the value of pursuing a scientific problem to its depths."

Another former Yanofsky postdoc, Bob Landick, in his nomination of Charley for an Abbott-ASM Lifetime Achievement Award, emphasized the value of mining in-depth rather than on the surface . "He dissected the pathway and regulation of tryptophan biosynthesis with such insight and imagination that he revealed major principles of biology that will remain forever etched in the edifice of science . Some of us spend our careers moving from one perceived opportunity to another, trying to be the first to uncover and glimpse a new gem of knowledge . Charles Yanofsky spent his working on a single large and ever more luminous stone . Through his perseverance, intellect, and creativity he polished it to reveal a jewel of unequaled clarity, each facet offering more insight into the truth of nature than dozens of more easily discovered gems . He truly is a giant of microbiology upon whose shoulders future generations will stand."

As we pondered over an appropriate present, Dale Oxender, who spent his sabbaticals with Charley, had the genial idea of asking the McCallum Company, orchid breeders, to name one of their creations "Charley." They declined—not dignified enough . Thus, did Phaleonopsis Charley Yanofsky receive its more respectable name . Laughter erupted when we learned that Charley Yanofsky is a cross between "Mad Lips" and "Stop Sign."

One of Charley's blooms graces the poster Mitzi designed to commemorate Gene Action '95 (Fig . 3) . After the poster presentation, Prasanta Datta brought out the real McCallum and set it on the table occupied by Charley and Edna . This living and lasting gift was possible because Dale knew that Edna and Irv's passion for orchids had infected Charley .

 
The response to our invitation was overwhelming and international . Charley's scientific progeny have dispersed around the globe . Former and current students, postdocs, technicians, colleagues, collaborators, friends, and relatives gathered (Fig . 4) . The respect, admiration, and love that poured forth were palpable . Old comrades were reunited, new acquaintances were made, and good feelings permeated the conference grounds .

 
The 10-min presentations were spaced out over 3 days to accommodate everyone who wanted to speak . Some talks were research oriented . Others, defying Charley's preference, exuded more humor and nostalgia than scientific insight . No one, however, summed up our thoughts and emotions more succinctly than George Bennett, who was a postdoc with Charley in the early seventies . He had found a "THANKS...CHARLEY" consensus sequence in the protein database . It has no tryptophan, but neither does the focus of so much of Charley's work: the tryptophan synthetase subunit . You see, Charley, I do remember something you taught me .

1. Babitzke, P., J . T . Stults, S . J . Shire, and C . Yanofsky. 1994 . TRAP, the trp RNA-binding attenuation protein of Bacillus subtilis, is a dodecameric protein that appears to recognize G/UAG repeats in the trpEDEFBA and trpG transcripts . J . Biol . Chem . 269:16597-16604.
2. Carbon, J., P . Berg, and C . Yanofsky. Missense suppression due to a genetically altered tRNA . Cold Spring Harbor Symp . Quant . Biol . 31:487-497.
3. Kelly, R . L., and C . Yanofsky. 1985 . Mutational studies with the trp repressor of E . coli support the helix-turn-helix model of repressor recognition of operator DNA . Proc . Natl . Acad . Sci . USA 82:483-487.
4. Madi, L., D . J . Ebbole, B . T . White, and C . Yanofsky. 1994 . Mutants of Neurospora crassa that alter gene expression and conidia development . Proc . Natl . Acad . Sci . USA 91:6226-6230.
5. Yanofsky, C. 1971 . Tryptophan biosynthesis in Escherichia coli . Genetic determination of the proteins involved . JAMA 218:1026-1035.
6. Yanofsky, C. 1981 . Attenuation in the control of expression of bacterial operons . Nature 289:751-758.
7. Yanofsky, C. 1984 . Comparison of regulatory and structural regions of genes of tryptophan metabolism . Mol . Biol . Evol . 1:143-161.
8. Yanofsky, C. 2001 . Advancing our knowledge in biochemistry, genetics, and microbiology through studies on tryptophan metabolism . Annu . Rev . Biochem . 70:1-37.
9. Yanofsky, C., B . C . Carlton, J . R . Guest, D . R . Helsinki, and U . Henning. 1964 . On the colinearity of gene structure and protein structure . Proc . Natl . Acad . Sci . USA 51:266-272.

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