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Home , Amita Sehgal, Michael Rosbash

Flies come through again,

Amita Sehgal

J Clin Invest. 2018;128(1):108-109. https://doi.org/10.1172/JCI97839.

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Jeffrey Hall, Michael Rosbash, and Michael Young have been awarded the 2017 in or Medicine for elucidating the molecular mechanism of the circadian (~24-hour) . Starting with the cloning of the period (per) gene, these investigators proceeded to demonstrate how endogenous clocks are composed of transcription-translation feedback loops (TTFLs) that drive rhythms in much of physiology. Beyond their fundamental significance for understanding normal physiology, the identification of clock mechanisms has led to a deeper appreciation of the health consequences of circadian disruption. Importantly, the Nobel prize–winning discoveries were made using a model, underscoring yet again the power of the fruit fly for biomedical research. Most physiological processes and behaviors display changes over a 24-hour period in synchrony with the day-night . Sleep, body temperature, metabolic activity, and secretion of many hormones, such as cortisol and insulin, represent but a few of the processes that are different in the morning versus the evening. These rhythms are driven by endogenous clocks that have a periodicity of approximately 24 hours, hence the name “circadian” from “circa” (about) and “dies” (day). Circadian rhythms are clearly an integral part of physiology, and disruptions in them are associated with a myriad of disorders, making it important to understand the mechanisms that generate them. A genetic basis for circadian clocks Although daily […]

Find the latest version: https://jci.me/97839/pdf VIEWPOINT The Journal of Clinical Investigation

Flies come through again, period

Amita Sehgal Program, Howard Hughes Medical Institute (HHMI), Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

Jeffrey Hall, Michael Rosbash, and above, usually does not precisely match the A mechanistic understanding Michael Young have been awarded the periodicity of the environment, providing of the clock 2017 Nobel Prize in Physiology or Medi- support for an internal rhythm–­generating The first insights into the clock mecha- cine for elucidating the molecular mech- process. A genetic basis for clocks was nism came from the finding, in the Hall anism of the circadian (~24-hour) clock. demonstrated by Bunning, whose work with and Rosbash laboratories, that per Starting with the cloning of the period bean plants in the 1930s demonstrated that (PER) and mRNA are expressed cyclically (per) gene, these investigators proceeded circadian period is heritable. Subsequently, (Figure 1). The surprise was that cycling of to demonstrate how endogenous clocks a historic screen for relevant genetic factors the mRNA appeared to depend on the pro- are composed of transcription-translation was conducted by Ron Konopka, a graduate tein, such that it was altered or lost in the feedback loops (TTFLs) that drive rhythms student in the laboratory of Seymour Ben- per missense or null mutants, respectively, in much of physiology. Beyond their fun- zer at Caltech. Konopka mutagenized fruit and rescue by functional PER could even damental significance for understanding flies (Drosophila melanogaster) and assayed drive cycling of the nonsense mutation– normal physiology, the identification of for X chromosome mutations that altered containing per01 transcript (4). This obser- clock mechanisms has led to a deeper circadian rhythms. He succeeded in identi- vation led to the model that per functions appreciation of the health consequences fying a fly line that had a short period (~19 in a feedback loop that could be an integral of circadian disruption. Importantly, the hours), a second with a long period (~29 part of the clock. Indeed, discovery of the Nobel prize–winning discoveries were hours), and yet another that lacked rhythms second gene (tim) made using a Drosophila melanogaster altogether. These mutations mapped to in the Young laboratory (5) substantiated model, underscoring yet again the power the same gene, which Konopka termed the the feedback loop mechanism and pro- of the fruit fly for biomedical research. period (per) gene (1). vided the basis for a clock model in which Most physiological processes and Isolation of the per gene awaited the the per and tim genes are coregulated and behaviors display changes over a 24-hour efforts of the trio that was recognized this cyclically expressed, and the two period in synchrony with the day-night fall. Using newly developed molecular negatively regulate transcription of both cycle. Sleep, body temperature, metabolic tools in the early 1980s, Jeffrey Hall and mRNAs. PER and TIM interact directly activity, and secretion of many hormones, Michael Rosbash of and display mutual regulation, most nota- such as cortisol and insulin, represent but and Michael Young of Rockefeller Univer- bly in ensuring nuclear expression of the a few of the processes that are different sity began the search for the per gene. Both heterodimer. Within the nucleus, they in the morning versus the evening. These groups localized per to a small region on the negatively regulate their own expression rhythms are driven by endogenous clocks X chromosome, and following extensive by inhibiting the activity of their tran- that have a periodicity of approximately 24 analysis of candidate transcripts, they suc- scriptional activators CLOCK (CLK) and hours, hence the name “circadian” from ceeded in pinpointing the relevant gene (2, CYCLE (CYC), discovered subsequently “circa” (about) and “dies” (day). Circadi- 3). Initial analysis of per did not provide reli- by the Hall and Rosbash groups. Timely an rhythms are clearly an integral part of able answers as to how it might be involved expression and activity of PER and TIM physiology, and disruptions in them are in timekeeping, but its isolation was nev- are critical for the appropriate regulation associated with a myriad of disorders, ertheless an enormous breakthrough, not of transcription and also depend on con- making it important to understand the only from a circadian point of view, but also trolled phosphorylation events. The Ros- mechanisms that generate them. from a behavioral biology perspective. Phe- bash laboratory showed that clock proteins notypes of the per mutants were found to are phosphorylated rhythmically, and the A genetic basis for circadian result from point mutations that led to ami- Young laboratory identified clock kinas- clocks no acid substitutions in the case of the short es, the first and best known being casein Although daily rhythms that persist in the and long alleles and to a nonsense codon kinase 1ε, which they termed doubletime absence of light-dark cycles had been noted that truncated the protein in the arrhythmic (dbt) and which phosphorylates and desta- in the 1700s, the idea that they are generat- allele. Thus, per became a paradigm for a bilizes PER. In fact, TIM counteracts the ed by endogenous clocks was not accepted behavioral gene, one in which even a single effect of DBT to stabilize PER. Synchrony until the 20th century. The circadian period nucleotide change could profoundly and of the clock with light, a key feature of cir- varies from species to species and, as noted specifically affect behavior. cadian clocks, is also accounted for by this clock mechanism, specifically by light-­ mediated degradation of the TIM protein

Conflict of interest: The author has declared that no conflict of interest exists. that consequently affects PER and the Reference information: J Clin Invest. 2018;128(1):108–109. https://doi.org/10.1172/JCI97839. entire loop (reviewed in ref. 6).

108 jci.org Volume 128 Number 1 January 2018 The Journal of Clinical Investigation VIEWPOINT

Figure 1. The molecular clock in Drosophila. In the major feedback loop of the Drosophila clock, the PER and TIM proteins inhibit activity of their transcriptional activators CLK and CYC at a specific time of day. Cycles of PER and TIM expression and activity are maintained through rhythmic transcription as well as through timely phosphorylation, degradation, and nuclear expression of the two proteins. Phosphorylation of PER by DBT (CK1ε) destabilizes it, an effect that is countered by TIM expression in the early night. Entrainment of the loop to light involves degradation of the TIM protein in response to signals transmitted by the (CRY) photoreceptor. In addition, the visual system can also entrain the clock. The figure is focused on the early findings that led to the awarding of this prize and does not include additional components, including interlocked loops, that are now known.

Importantly, a negative feedback loop again first made in flies by the prize winners, Address correspondence to: Amita Seh- consisting of two coregulated genes is now that clock genes are expressed not only in gal, 10-136 Smilow Research Center, the prevailing model for all animal clocks, the brain, but also in other organs, has led 3400 Civic Center Blvd., Bldg. 421, Phil- and most of the molecular components are to the concept of a circadian system that adelphia, Pennsylvania 19104-5158, USA. also conserved from flies to mammals. In consists of multiple synchronized clocks Phone: 215.573.2985; Email: amita@ fact, the Clock gene was identified first in across the organism. In any given organ, as pennmedicine.upenn.edu. mice by the laboratory of Joseph Takahashi many as 15% to 20% of the transcripts may 1. Konopka RJ, Benzer S. Clock mutants of Dro- (7), and in the past two decades molecular cycle under clock control, and they are typ- sophila melanogaster. Proc Natl Acad Sci U S A. circadian research in Drosophila and mam- ically specific for the functional output of 1971;68(9):2112–2116. mals has moved in parallel. The signifi- that organ. Loss of synchrony among body 2. Zehring WA, et al. P-element transformation cance of the fly work cannot, however, be clocks, or of endogenous clocks to the envi- with period locus DNA restores rhythmicity to overstated. The molecular lesion in the first ronment, has deleterious physiological con- mutant, arrhythmic Drosophila melanogaster. Cell. 1984;39(2 pt 1):369–376. characterized human circadian disorder sequences. Given that circadian desynchro- 3. Bargiello TA, Jackson FR, Young MW. Res- was traced to the Per2 gene (mammals have ny/disruption — caused by jet lag, shift work, toration of circadian behavioural rhythms 3 period genes), specifically to phosphory- constant exposure to light, or even mistimed by gene transfer in Drosophila. Nature. lation of PER2 by casein kinase 1ε (8), and feeding — is quite common in modern soci- 1984;312(5996):752–754. subsequent human studies have also been ety, it could be a significant contributor to 4. Hardin PE, Hall JC, Rosbash M. Feedback of the Drosophila period gene product on circadian informed by clock mechanisms identified pathological conditions, which is supported cycling of its messenger RNA levels. Nature. in Drosophila. by findings of circadian abnormalities during 1990;343(6258):536–540. aging and in metabolic, cardiovascular, neu- 5. Sehgal A, Price JL, Man B, Young MW. Loss Conclusions rological, neuropsychiatric, and other dis- of circadian behavioral rhythms and per RNA Our current understanding of clocks now eases. Recognition of the circadian field by oscillations in the Drosophila mutant timeless. Science. 1994;263(5153):1603–1606. involves many added layers of complexi- the Nobel Assembly at Karolinska Institute is 6. Dubowy C, Sehgal A. Circadian rhythms and ty, including additional connected loops, timely and will undoubtedly focus attention sleep in Drosophila melanogaster. . but the basic principle described above on these daily cycles and their importance 2017;205(4):1373–1397. can explain circadian regulation in gener- for health and disease pathogenesis. 7. Vitaterna MH, et al. Mutagenesis and mapping al. Transcriptional effects of clock proteins of a mouse gene, Clock, essential for circadian behavior. Science. 1994;264(5159):719–725. are not restricted to their own mRNAs, but Acknowledgments 8. Toh KL, et al. An hPer2 phosphorylation site apply to many genes, thereby driving rhyth- Work in the laboratory is supported by mutation in familial advanced sleep phase syn- mic expression of those genes. The finding, NIH grant R37NS048471. drome. Science. 2001;291(5506):1040–1043.

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