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Cis-Combination of the Classic per^S and per^L Mutations Results in Arrhythmic Drosophila with Ectopic Accumulation of Hyperphosphorylated PERIOD Protein

Graduate Program in Physiology and Neurobiology, Rutgers University, Center for Advanced Biotechnology and Medicine, Piscataway, NJ

Suzanne DiMassa

Rutgers University, Center for Advanced Biotechnology and Medicine, Piscataway, NJ

Eun Young Kim

Rutgers University, Center for Advanced Biotechnology and Medicine, Piscataway, NJ, Department of Life Science, Yonsei University, Wonju, Gangwon-do, South Korea

Kiho Bae

Department of Life Science, Yonsei University, Wonju, Gangwon-do, South Korea

Isaac Edery

Department of Molecular Biology and Biochemistry, Rutgers University, Center for Advanced Biotechnology and Medicine, Piscataway, NJ, Graduate Program in Physiology and Neurobiology, Rutgers University, Center for Advanced Biotechnology and Medicine, Piscataway, NJ, edery{at}cabm.rutgers.edu

The 1st circadian "clock" gene identified was the X-linked period (per) gene in Drosophila melanogaster. In the pioneering initial report, Konopka and Benzer (1971) characterized 3 alleles of per that shortened (per ^S; ~19 h), lengthened (per ^L; ~29 h), or abolished (per ⁰) circadian behavioral rhythms. They also showed that transheterozygotes carrying the per ^S and per ^L mutations exhibit robust behavioral rhythms with nearly normal periods of ~23 h, highlighting the semidominant nature of many clock mutants. In this study, per ⁰ flies bearing a doubly mutated per transgene that carries both the per ^S and per ^L alleles (per ⁰; per ^S/L) were analyzed for behavioral and molecular rhythms. Unlike singly mutated versions, the per ⁰;per ^S/L transgenic flies are arrhythmic in constant dark conditions and exhibit little, if any, entrainment to daily light—dark cycles. In a wildtype per ⁺ background, expression of per ^S/L abolishes behavioral rhythms, indicating that it functions in a transdominant negative fashion. Biochemical analysis of head extracts revealed that only hyperphosphorylated isoforms of the PERS/L protein are detected throughout a daily cycle, and the levels remain constant. Intriguingly, little if any PERS/L is observed in key pacemaker neurons that control daily activity rhythms, consistent with the notion that hyperphosphorylated isoforms of PER are unstable. Nonetheless, PERS/L is detected in ectopic cells in the brain, in which it exhibits an unusual localization, mainly staining the periphery of the nucleus. These results suggest that posttranslational mechanisms play a key role in limiting the accumulation of PER to specific cells. On a broader scope, our results indicate that the semidominant effects of period-altering alleles observed in trans are not necessarily preserved in the cis-configuration and that novel phenotypes can emerge.

Key Words: Drosophila • circadian rhythms • period protein • phosphorylation • perS • perL

References

• Bae K. and Edery I. (2006) Regulating a circadian clock's period, phase and amplitude by phosphorylation: insights from Drosophila . J Biochem ( Tokyo ) 140: 609-617 .[Abstract/Free Full Text]
• Baylies MK, Bargiello TA, Jackson FR, and Young MW (1987) Changes in abundance or structure of the per gene product can alter periodicity of the Drosophila clock . Nature 326: 390-392 .[CrossRef][Medline] [Order article via Infotrieve]
• Baylies MK, Vosshall LB, Sehgal A., and Young MW (1992) New short period mutations of the Drosophila clock gene per . Neuron 9: 575-581 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Chang DC and Reppert SM (2003) A novel C-terminal domain of Drosophila PERIOD inhibits dCLOCK:CYCLE-mediated transcription . Curr Biol 13: 758-762 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Curtin KD, Huang ZJ, and Rosbash M. (1995) Temporally regulated nuclear entry of the Drosophila period protein contributes to the circadian clock . Neuron 14: 365-372 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Darlington TK, Wager-Smith K., Ceriani MF, Staknis D., Gekakis N., Steeves TD, Weitz CJ, Takahashi JS, and Kay SA (1998) Closing the circadian loop: CLOCK-induced transcription of its own inhibitors per and tim . Science 280: 1599-1603 .[Abstract/Free Full Text]
• Edery I., Rutila JE, and Rosbash M. (1994a) Phase shifting of the circadian clock by induction of the Drosophila period protein . Science 263: 237-240 .[Abstract/Free Full Text]
• Edery I., Zwiebel LJ, Dembinska ME, and Rosbash M. (1994b) Temporal phosphorylation of the Drosophila period protein . Proc Natl Acad Sci U S A 91: 2260-2264 .[Abstract/Free Full Text]
• Gekakis N., Saez L., Delahaye-Brown AM, Myers MP, Sehgal A., Young MW, and Weitz CJ (1995) Isolation of timeless by PER protein interaction: defective interaction between timeless protein and long-period mutant PERL . Science 270: 811-815 .[Abstract/Free Full Text]
• Hall JC ( 2003) Genetics and molecular biology of rhythms in Drosophila and other insects . Adv Genet 48: 1-280 .[Web of Science][Medline] [Order article via Infotrieve]
• Hamblen-Coyle MJ, Wheeler DA, Rutila JE, Rosbash M., and Hall JC (1992) Behavior of period-altered circadian rhythm mutants of Drosophila in light:dark cycles (Diptera: Drosophilidae) . J Insect Beh 5: 417-446 .[CrossRef]
• Hardin PE (2005) The circadian timekeeping system of Drosophila . Curr Biol 15: R714-R722 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Hardin PE, Hall JC, and Rosbash M. (1990) Feedback of the Drosophila period gene product on circadian cycling of its messenger RNA levels . Nature 343: 536-540 .[CrossRef][Medline] [Order article via Infotrieve]
• Helfrich-Forster C. (2005) Neurobiology of the fruit fly's circadian clock . Genes Brain Behav 4: 65-76 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Houl JH, Yu W., Dudek SM, and Hardin PE (2006) Drosophila CLOCK is constitutively expressed in circadian oscillator and non-oscillator cells . J Biol Rhythms 21: 93-103 .[Abstract/Free Full Text]
• Huang ZJ, Edery I., and Rosbash M. (1993) PAS is a dimerization domain common to Drosophila period and several transcription factors . Nature 364: 259-262 .[CrossRef][Medline] [Order article via Infotrieve]
• Kaneko M. and Hall JC (2000) Neuroanatomy of cells expressing clock genes in Drosophila: transgenic manipulation of the period and timeless genes to mark the perikarya of circadian pacemaker neurons and their projections . J Comp Neurol 422: 66-94 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Kim EY and Edery I. (2006) Balance between DBT/CKIepsilon kinase and protein phosphatase activities regulate phosphorylation and stability of Drosophila CLOCK protein . Proc Natl Acad Sci U S A 103: 6178-6183 .[Abstract/Free Full Text]
• Ko HW, Jiang J., and Edery I. (2002) Role for Slimb in the degradation of Drosophila Period protein phosphorylated by Doubletime . Nature 420: 673-678 .[CrossRef][Medline] [Order article via Infotrieve]
• Konopka RJ and Benzer S. (1971) Clock mutants of Drosophila melanogaster . Proc Natl Acad Sci U S A 68: 2112-2116 .[Abstract/Free Full Text]
• Konopka RJ, Pittendrigh C., and Orr D. (1989) Reciprocal behaviour associated with altered homeostasis and photosensitivity of Drosophila clock mutants . J Neurogenet 6: 1-10 .[Medline] [Order article via Infotrieve]
• Lee C., Bae K., and Edery I. (1998) The Drosophila CLOCK protein undergoes daily rhythms in abundance, phosphorylation, and interactions with the PER-TIM complex . Neuron 21: 857-867 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Lyons LC, Darlington TK, Hao H., Houl J., Kay SA, and Hardin PE (2000) Specific sequences outside the E-box are required for proper per expression and behavioral rescue . J Biol Rhythms 15: 472-482 .[Abstract/Free Full Text]
• Majercak J., Chen WF, and Edery I. (2004) Splicing of the period gene 3'-terminal intron is regulated by light, circadian clock factors, and phospholipase C . Mol Cell Biol 24: 3359-3372 .[Abstract/Free Full Text]
• Marrus SB, Zeng H., and Rosbash M. (1996) Effect of constant light and circadian entrainment of perS flies: evidence for light-mediated delay of the negative feedback loop in Drosophila . Embo J 15: 6877-6886 .[Web of Science][Medline] [Order article via Infotrieve]
• Myers MP, Wager-Smith K., Rothenfluh-Hilfiker A., and Young MW (1996) Light-induced degradation of TIMELESS and entrainment of the Drosophila circadian clock . Science 271: 1736-1740 .[Abstract]
• Rothenfluh A., Abodeely M., and Young MW (2000) Short-period mutations of per affect a double-time-dependent step in the Drosophila circadian clock . Curr Biol 10: 1399-1402 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Rutila JE, Edery I., Hall JC, and Rosbash M. (1992) The analysis of new short-period circadian rhythm mutants suggests features of D. melanogaster period gene function . J Neurogenet 8: 101-113 .[Web of Science][Medline] [Order article via Infotrieve]
• Sidote D., Majercak J., Parikh V., and Edery I. (1998) Differential effects of light and heat on the Drosophila circadian clock proteins PER and TIM . Mol Cell Biol 18: 2004-2013.
• Tomioka K., Sakamoto M., Harui Y., Matsumoto N., and Matsumoto A. (1998) Light and temperature cooperate to regulate the circadian locomotor rhythm of wild type and period mutants of Drosophila melanogaster . J Insect Physiol 44: 587-596 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Vanselow K., Vanselow JT, Westermark PO, Reischl S., Maier B., Korte T., Herrmann A., Herzel H., Schlosser A., and Kramer A. (2006) Differential effects of PER2 phosphorylation: molecular basis for the human familial advanced sleep phase syndrome (FASPS) . Genes Dev 20: 2660-2672 .[Abstract/Free Full Text]
• Wheeler DA, Hamblen-Coyle MJ, Dushay MS, and Hall JC (1993) Behavior in light-dark cycles of Drosophila mutants that are arrhythmic, blind, or both . J Biol Rhythms 8: 67-94 .[Abstract/Free Full Text]
• Yildiz O., Doi M., Yujnovsky I., Cardone L., Berndt A., Hennig S., Schulze S., Urbanke C., Sassone-Corsi P., and Wolf E. (2005) Crystal structure and interactions of the PAS repeat region of the Drosophila clock protein PERIOD . Mol Cell 17: 69-82 .[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
• Yu Q., Jacquier AC, Citri Y., Hamblen M., Hall JC, and Rosbash M. (1987) Molecular mapping of point mutations in the period gene that stop or speed up biological clocks in Drosophila melanogaster . Proc Natl Acad Sci U S A 84: 784-788 .[Abstract/Free Full Text]
• Zeng H., Qian Z., Myers MP, and Rosbash M. (1996) A light-entrainment mechanism for the Drosophila circadian clock . Nature 380: 129-135 .[CrossRef][Medline] [Order article via Infotrieve]
• Zerr DM, Hall JC, Rosbash M., and Siwicki KK (1990) Circadian fluctuations of period protein immunoreactivity in the CNS and the visual system of Drosophila . J Neurosci 10: 2749-2762 .[Abstract]
• Zordan M., Osterwalder N., Rosato E., and Costa R. (2001) Extra ocular photic entrainment in Drosophila melanogaster . J Neurogenet 15: 97-116 .[Web of Science][Medline] [Order article via Infotrieve]

Journal of Biological Rhythms, Vol. 22, No. 6, 488-501 (2007)
DOI: 10.1177/0748730407306929


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