Abstract
Seymour Benzer (left) and Ron Konopka, together at CalTech in 2003, catalyzed by the visit of Joel Levine to Pasadena. Photo courtesy of Joel Levine (now at the University of Toronto Mississauga).
The Kapusta Kid (TKK) has died. That obscure nickname was given to himself by Ronald J. Konopka, chrono-geneticist extraordinaire. Ron died February of this year. This did not shock me, for he was in poor health last time we got together (fall 2012). Furthermore, TKK did not take the greatest care of himself, after he retired prematurely during 1990 (when he was only 43). This preamble gets ahead of the deal, or could provoke head-scratching as to “why an obituary for someone who disappeared from formal biology 25 years ago?”
Well, Dr. Konopka did not truly disappear, having not done with respect to lingering research-related importance. In fact, I gingerly suggest that he, as a rhythm investigator, will never be forgotten. This is because TKK—who was of Polish extraction and invoked a jocular appellation for himself to include that language’s word for “cabbage”—was the seminal chronobiologist who infused definitive genetics into studies of rhythm regulation, in general, and as a possible investigative inroad toward understanding the circadian clock, in particular.
RJK started it all, chronogenetically, with respect to an enterprise that by now has ballooned to industrial-level research. He did so by generating single-gene rhythm mutants, from scratch. He did so (in turn) by systematically searching for heritable variants displaying conspicuous subnormalities or anomalies of circadian rhythmicity. Ron initiated this activity by exploiting the generic “biogenetic” potential of Drosophila. All of this began to well up (rhythm-wise) during the historic summer of 1968, when TKK was not that much more than a kid. Nonetheless, Ron realized, as the final third of the 20th century began to unfold, that a handful of fruit fly folk were expanding their genetic horizons: no longer studying inheritance alone (genetic variants as tools for elucidating “the basics” of genic and chromosomal heredity in metazoans) but instead beginning to take genetic approaches potentially to achieve some extra understanding of various biological processes. These included development (heavily involved at the dawn of biogenetics) as well as functioning of mature animals (a minor subenterprise during those early days). One of the 1960s pioneers was the fledgling behavior-geneticist Seymour Benzer, in whose lab at the California Institute of Technology Ron Konopka researched as a grad student.
This professional situation entailed right place/right time, insofar as kick-starting chronogenetics was concerned. It is most important to note that RJK had a longstanding interest in circadian rhythmicity before he joined the Benzer lab. Also well before that time, the famed lab of Thomas Hunt Morgan (THM) had established itself as a “Drosophila center,” starting at Columbia University, moving to CalTech about one-third into the 20th century. For Benzer’s part, it seemed a nonrandom choice for him metaphorically to move into flies at the latter location, when that former prokaryotic geneticist physically moved from Purdue University to Pasadena. This piece of science history is probably well known, but it also connects with more specific, ostensibly smaller, features of the mid-1960s situation; these were important in terms of how they promoted biogenetic pioneering.
Thus, Ed Lewis—a direct descendant of THM and co-professor with Benzer at CalTech—had innovated a powerful plus easily applied method aimed at increasing frequencies of new mutations in Drosophila: chemical mutagenesis. Within another lab there, Ed’s technical accomplishment allowed Seymour himself, then Ron, readily to effect mutant hunting and to do so in ways that novel variants would not be impractically rare. It bears mentioning on the side that Konopka and Lewis became solid, mutually respecting colleagues during the former’s training stint (resulting eventually in RJK earning his PhD during 1972). Actually, this obituary module is well worth entering, because Ron’s seminal achievements as a chronogeneticist were closely connected with how good he was at handling fruit fly genotypes (above and beyond whatever rhythm-related phenotype was at hand). Ed Lewis, one of the past masters at dealing with Drosophila genetics of the hardest-core variety, became a kind of informal genetics mentor for Konopka, although it was also necessary for TKK to be willing and able to soak up the requisite principles and practices, therefore to establish himself as a deeply expertised geneticist (extending beyond what he brought with him to CalTech, which again was an abiding interest in matters rhythmic).
Soon after Ron and I became labmates (1971), I registered that he was one of the best Drosophila geneticists around. We began to overlap in Benzer’s lab shortly before the announcement of remarkable Drosophila mutants burst forth: Konopka and Benzer (1971). In retrospect, this is one of the most important publications in chronobiological history. The paper was entitled “Clock Mutants in Drosophila melanogaster.” Why such an assertive title (vis-à-vis its first word)? This harks back a short prepublication distance to when Ron effected his first-blush mutagenesis and screening for genetic variants. He managed to identify his original rhythm mutant by the end of the aforementioned summer of 1968. The variant was aperiodic with regard to the emergence of adult fruit flies from metamorphosis, against a background of genetically normal cultures of Drosophila “eclosing” via daily peaks near dawn. Ron, a fan of circadian rhythms, was well aware of this periodic phenomenon, which had long been studied (pre-late-1960s) in Drosophila pseudoobscura. The latter species was not a good “genetic system” (and remains unexploitable in that sphere).
An additional reason for me to regard Konopka as a salient member of the Benzer lab, at the time I joined that group as a wet-behind-ears postdoc, was connected to my “extra” appreciation of his thesis-based accomplishments—including their conspicuous chrono-component. This was because, as a kind of college-days fluke far away from California, I had learned about circadian rhythms “in pseudoobscura.” This was because of course lectures presented by a young instructor recently emerged (ca. 1966) from the lab that had been studying “morning peaks” of such emerging flies.
In order to make eclosion rhythmicity potentially analyzable genetically, RJK started by mutagenizing D. melanogaster flies (à la Lewis), leading to establishment of 200 substrains (again during that fateful late 1960s summer). Ron observed each culture by eye, wondering whether he could discern eclosion anomalies. Accordingly, he manifested fine expertise in terms of the relevant rhythmic trait (more than his preestablished interest in daily cycles). Thereby, he identified one “sport” (out of the original 200), for which adult emergences seemed abnormally spread out across a given 24-h day. This led Ron to formulate Konopka’s First Law: “If you don’t find it among the first 200, quit.” He would deadpan that phrase, prompting listeners to ask, “What’s your Second?” (Answer, naturally: “There is none.”) It seemed to me that this kind of guerilla theater was deeply ingrained in RJK’s nature, going back to his formative years in Ohio (and undergrad studies at Dayton University). Silly as the following may seem, I was drawn to Ron’s sense of humor and entertaining intralab antics, as well as to his 2-fold investigative astuteness. I also registered that his biointerests went way back, rhythm-wise and other-wise. For example, Ron was one of those lepidopteran guys who had long been scoping out butterflies and moths in the wild (capture, pin-up, etc.). Years later, I was pleased to introduce The Kapusta Kid to another rhythm person: an investigator of mammals who had recently entered the chronogenetic subfield, and had long been a lepidopteran aficionado. Those 2 fellows, who had only heard of one another research-wise, seemed meaningfully to connect—better than with regard to the purposes of the gathering (this interaction occurred during some dumb meeting, for which I had been authorized to invite participants and made sure to include the mammalian guy along with the one who indirectly made chronogenetics devoted to that “relevant” animal what it is today).
In further context of straying from matters Konopka, other than those connected with his chronobiology, I cannot help mention that I visited him many times (beyond the micro-epoch when we were labmates). This entailed me making pilgrimages to Upstate New York. Ron had been exiled to there (Clarkson University), after his first faculty-level stint (back at CalTech, ending around the early 1980s). When he was outrageously denied tenure at Clarkson and gave up running round academic circles, he retired to his beloved Pasadena. I visited him there several times, making me wonder whether I was merely paying homage. No, because I realized such visits were pleasant, at least in part because of our common interests: rock ‘n’ roll music, movies, sports, beer drinking, and profanity (Ron at times was a dirty, dirty man). His funky nature was signified further by playing the ponies (I guess he had plenty of time to do so). More than once The Kid took me to the Santa Anita racetrack, where we spent pleasant and interesting afternoons (Ron, who had come to know what he was doing, would win five bucks; I would lose about that amount).
In any case, who wouldn’t want to reconnect with someone who was a flat-out good guy? Speaking of (also) interesting, most of my visits entailed Ron and guest-including forays into other parts of the Los Angeles “basin,” including what he called “LPR” outings into the San Gabriel mountains (that abbreviation being Ron’s shorthand for “Low Probability of Return”). Well, we not only survived such hikes, but they were intellectually appealing as well as enjoyable. These of our recreational activities made me realize what a superb zoologist and botanist was The Kid (also registered by me when he took me on treks through the “boreal forest,” south of Potsdam New York in the Adirondacks). No wonder RJK was so good at investigating the biological side of his system, never aiming to impress solely in terms of how good he was at all the Drosophila-genetic bibble-dee-boo.
That Dr. Konopka was more than good at all those things became noticeable all the way back to his first stint at CalTech. After inducing, identifying, and solidly establishing his first circadian rhythm variant, the “arrhythmic” sport, he continued to mutagenize and screen. This led to Ron’s remarkable recovery of 2 additional variants: one whose cycle durations were only ~20 h (in constant conditions) and a third type for which cycle durations were ca. 5 h longer than normal. He found all 3 such variants by brute-force examination of eclosing fruit fly cultures. Subsequently—as and coreported within his 1971 paper to reflect a rhythm-related innovation technically—Ron showed that automated single-fly monitoring of adult behavior revealed the same kinds of circadian defects-cum-anomalies: free-running locomotor cycles governed by a fast clock, a slow one, or no clock as the case may be. What that paper reported back then, in terms of assessing and analyzing behavioral rhythmicity, became de rigeur for vast numbers of subsequent investigators, who followed Ron’s lead into the fly rhythm business.
Meanwhile—late 1960s/early 1970s—what luck for the seminal chronogeneticist: all 3 circadian-rhythm categories worth of abnormalities that one could have surmised a priori. Perhaps it was lucky that anything came out of Ron’s mutant hunting. Obviously, however, his “numerator” was >0, and by the way, the denominator for RJK’s series of screens was ca. 1900 substrains descending from the initial mutagenizees. The latter number implies lots of hard work and stick-to-itive-ness. More and better: Konopka did not come across 3 sports out of 30 tries or so (cf. Konopka’s First Law), also not something like 200 out of ca. 2K. If the penultimate number just entered had resulted: what to follow up among an avalanche of new variants? And might triple-figures’ worth of new mutants suggest that all kinds of genes can “get-to the clock” and foul up its operation in indirect, probably uninformative ways? Ron was aware of this matter, gleaned by me as we caroused within his hovel (South Santa Anita Ave, Pasadena) through the 1990s and into the current century: musing about the old days from time to time, with astute TKK being well aware of where the principles and practices pioneered by him had gone (post-1970s, including after he retired): “Gee,” he reflected, “I was glad to have found something [from all that screening], and it was even better that that tractable trio of mutants stimulated sharp focus on a ‘clock gene’ [one that can mutate to speed up or slow down the circadian clock’s pace; not, when mutated, solely eliminate rhythmicity because of developmental brain damage or the like].”
What am I talking about here, if anything, and in context of errant retrospective musings? Bringing this science-based obit back to why the subject’s pioneering may warrant presentation of such a doc: the early 1970s chronogenetic situation got way better than a matter of successful mutant hunting, when RJK began to dig deeply into etiologies of the 3 kinds of rhythm-abnormal strains. Fearlessly—unlike genetically related attitudes I have noticed over many years of teaching undergrads and of interacting with peers—RJK determined that each of his trio of variant types was accounted for by single-gene mutations. So demonstrating is straightforward, but exercising the requisite elementary genetics has not infrequently been blown off or botched in biogenetic history. Not so for The Kid, at least in part because Ed Lewis was around at the time (the latter having been a student of A. H. Sturtevant, son of THM, within whose lab “how to map a mutation and do more than apply the principles of Mendelian segregation” had been famously established). For Ron’s part: not only one genetic locus mutated within each of his 3 rhythm-abnormal strains but also that only one locus on one chromosome happened to be involved every time (what luck!). To repeat: this one locus—which Ron named period, one of the most interesting categories of genes known “in organisms”—can be mutated to cause circadian clock-pace anomalies or to eliminate Drosophila’s inborn ability to manifest daily biocycles.
It bears entering as well that Ron’s genetic professionalism—to prove “one gene per variant type and only one involved in all 3 of the variations”—extended beyond basic segregational plus meiotic-recombinational characterizations. Also, he was aware of a necessity—and how to interpret the results of—“putting a given mutation over another of the 3” (complementation tests, which have flummoxed other biogeneticists). Even more and better: how valuable it can be to appreciate the power of chromosome aberrations, application of which can nail down a given instance of genetic etiology to a highly resolved subset of the relevant chromosome. For this, Konopka thoroughly understood the potential “power” and once again fearlessly acquired → applied relevant strains of D. melanogaster (preexisting aberrations, classic ones even, within a genotypic category that causes “some” to recoil in incomprehending terror).
So—with respect (much) to Konopka and Benzer (1971)—what a superbly reported set of research accomplishments, having to do with 2 unrelated types of lab work. RJK was therefore special, not only in terms of the novel variants he laboriously searched for and found but also how he dealt with them at a level extending beyond the purely chronobiological arena. I, as an alum of Benzer’s lab, am aware of every single paper that came out of it (from the 1960s into the first decade of the current century). The one that Seymour coauthored in 1971 was and is (a) the most inherently impressive and interesting element of Benzer’s entire oeuvre and (b) a paper that arguably had the most heuristic value.
Such “value” speaks to ways that chronogenetic concerns got extended hugely, with the Drosophila business, and how “genetic approaches to the clock” came to insinuate themselves into chronostudies of every imaginable type of organism here on The Third Rock (notably the fungus Neurospora crassa as well as various mammalian types, including us). Proceeding from that arch claim, here are elements of what The Kapusta Kid accomplished on behalf of the fruit fly side, after his seminal publication burst forth: not only “here are some remarkable rhythm-defective mutants” but also “here is how they came about, including hi-res localization of the [3] genetic variants.”
In part by wallowing in semi-personal concerns of this obit author, he now cuts to the “chrono-molecular-genetic” chase. This connects with how the lab of yours truly and a pair of additional Drosophila groups came to exploit Konopka and Benzer (1971). Ron’s discovery of rhythm variants, superbly augmented by his establishing the genotypic side, set the stage for other research groups indeed to identify his period gene at the DNA level. One such group included me, ensconced within a small university near Boston. How the Hall lab, and that supervised by Michael Rosbash, got into the rhythm biz prompts me to divulge additional elements of Konopka-related funk. In this regard, once I and colleagues (toiling within that New England institution) sensed we had isolated fragments of DNA that had to at least be near Konopka’s gene, we aimed physically to introduce such genomic material into Drosophila strains whose only form period was (Ron’s) arrhythmia-inducing perZero “allele.” (Don’t ask, you who are afraid of even basic genetics, what the quoted noun means.) Our hope was to determine which if any such incoming molecules would “rescue” effects of that per0 mutation. We so introduced and generated preliminary chrono-bio info as to effects of certain such DNA molecules. But we were not locally convinced or confident. So, I asked Ron Konopka if I—accompanied by a labmate—could vote with our feet: travel to where he had come to work, aforementioned Upstate New York (TKK having arrived there a bit before that time, which by then was mid-1980s in terms of our New England enterprise-ette). Why such a requested visit? So that Ron could apply his well-established wherewithal to monitor behavioral rhythmicity of Drosophila individuals, then interface the pertinent digital data with analytical “programs,” optimally to determine whether a given adult fly had behaved rhythmically (and, if so, with what cycle duration). At that time, with me knowing a fair amount about Ron’s abilities and attitudes, I was unsurprised that his response was “Sure, come.” I and an associate arrived (in Potsdam, NY) some dark night, at which time we encountered a substantially wasted Kapusta Kid. Soon, the 2 visitors caroused to get into that state. Next morning, somehow the 3 of us were functional enough to drive to Ron’s meager lab (part of the afore-noted university in Potsdam, NY, unknown except with regard to its hockey team). Our host was preaware that that pair of visitors was bringing arbitrarily labeled containers of adult Drosophila, whereby the former was willing and exquisitely able to test them for locomotor rhythmicity. Ron was so set up, by virtue of applying devices plus expertise that he had brought from far away (CalTech → Clarkson). We 2 visitors utterly believed that Ron would come through, in terms of his attitude and acumen (the latter factor being way better than what was extant then at our home base). The fly containers we left with Dr. Konopka were labeled in coded manners: no indicators as to what type of per0 + DNA-infused types were at hand (or whether certain tubes contained per0-only controls). Unsurprising to me, Ron was more than OK with taking in such research materials in that context. Outcome: approximately 1 month later, he transmitted eastward the numerical results of his many flies’ worth of tests. When we at home decoded the genotypic designators, we knew that we as part of a 3-lab group had identified pieces DNA with rhythm-promoting period-based activity (that trio of labs encompassed 2 within New England plus Ron’s at Clarkson, such as it was).
So, Ronald J. Konopka saved our ass, in terms of us legitimately reporting (late 1984) that we had “cloned the period clock gene,” buttressed by identifying it at the level of bioassaying putative per DNA. I could never forget what he did for us as relative neophytes in the field, way back then. That was when chronobiology going molecular—molecular-genetic really—poised elements of the overall field to proceed toward making meaningful insights into “what is a circadian clock,” beyond and deeper than formalistic understandings (e.g., organisms manifest clock-controlled 24-h rhythms in the absence of daily earth cycles, and one can deeply analyze the “phase-shifting” effects of environmental stimuli on circadian rhythmicity).
So, Ronald J. Konopka set said field on a path potentially to “crack the clock case.” He took his first steps along it during the late 1960s and continued to traverse it well into the 1980s. During the former micro-epoch, his period mutants, whose phenogenetics would not quit, actually opened the investigative pathway. This was so even though it took more than a dozen years to make Ron’s first-blush accomplishments proceed to the “concrete” level. (What is the period gene in addition to “a locus” that can mutate to cause fascinating rhythm phenotypes?)
That I and colleagues were able to “go molecular” was materially contributed to by Ron’s actual lab work, underpinned by his expertise and optimistic generosity. This was near the beginning of a ~10-year period during which chronobiology began to take off, at least in part because of the molecular-genetic augment. As the second half of the 1990s unfolded, the case cracking in question spread out and deepened into matters rhythmic related to many other kinds of organisms (even into the microbial world as it dips below the eukaryotic/prokaryotic barrier). In this regard, it is difficult to avoid inserting some words as to the specific consequences of per going molecular. It had to do if there would be potential for full understanding of the deal. Yet it could never have gone had RJK not pointed to the gene via his spectacular mutations, followed by increasingly high-res determination of where period is located. This allowed for it (as for other famous genes in biogenetic history) to be isolated, quintessentially according to “where within the genome” (no a priori clue as to what per encodes). Owing to the premature end of his career, Konopka did not participate in secondary and tertiary analyses of “what’s the per product doing?” This points to the fact that that gene is expressed (RNA, protein) in a way that defines a daily molecular rhythm—the same kind of temporally modulated expression patterns manifested by several subsequently identified clock genes (microbes → → mammals). These discoveries about “cycling” molecules were made in advance of figuring out just what a given clock protein (final gene product) is doing at the cellular level, although much is known in that arena, too, by now.
None of this would have happened, absent what Ron Konopka pushed first. See why I claim how truly seminal a paper was Konopka and Benzer (1971)? The findings presented back then were not destined to be so crucially prescient. But it worked out. And all of this stemmed from RJK’s hard work, intellectual courage, and rhythm-rooted motivation. I myself have never interacted with anyone who was “so good” at both chronobiology and high-level genetics.
I will close by putting forth, with no compunction, admiration-based words of the large-scale variety . . . about The Kapusta Kid (RIP). To start, however, with something of a sour note: that he died recently will understandably lead to some contemporary shrugs. After all, his groundbreaking work began to be performed almost 50 years ago. Then, during the second half of that time, Ron disappeared into too-early retirement. Yet in light of how the chronobiological enterprise has molecularly exploded over the course of the ~2 most recent decades: that one can get into this field and sustain a career within it goes back to one key guy. Oh so many of us (fruit fly folk and large numbers of others investigating on behalf of noninvertebrate “systems”) would barely have had jobs had it not been for what Ronald J. Konopka formulated and dramatically got done as of two-thirds into the previous century. Had he not so innovated and achieved, late 20th/early 21st-century careers of many would not have been as successful and interesting as they seem to have been. We might not even have been able to support our basic existences (families, mortgage payments, etc.)—again, if The Great Ron Konopka had not been willing and quintessentially able to accomplish when he did do. More than incidentally, that was at a time when chronobiology was an occult quirky kind of basket case in the minds of many. Immediately after the first rhythm meeting I attended during 1983, a high-level organizer addressed the assembled chrono-folk to scold us about the “last such meeting we will support, if recently displayed rhythm-related folderol doesn’t get better, soon.”
Yet, with thanks significantly to now-departed Ronald J. Konopka, the “clock field” in question started materially to mature. Then it soared upward to where it is now: circadian clocks are no longer an obscure mystery or miracle, whether or not such biological timers are important components of just about every life form here on our planet. They are, because these features of life thereon are as crucial for organismal “lifestyles” as can be, let alone being an enduringly interesting aspect of life per se (perhaps I should have closed with “per se”).