β-TrCP1-Mediated Degradation of PERIOD2 Is Essential for Circadian Dynamics

Abstract

Regulated degradation of circadian clock proteins is a crucial step for rhythm generation per se but also for establishing a normal circadian period. Here, the authors show that the F-box protein beta-transducin repeat containing protein 1 (β-TrCP1) as part of the E3 ubiquitin ligase complex is an essential component of the mammalian circadian oscillator. Down-regulation of endogenous β-TrCP1 as well as expression of a dominant-negative form both result in lengthening of the circadian period in oscillating fibroblasts. These phenotypes are due to an impaired degradation of PERIOD (PER) proteins, since expression of β-TrCP interaction-deficient PER2 variants—but not wild-type PER2—results in a dramatic stabilization of PER2 protein as well as in the disruption of circadian rhythmicity. Mathematical modeling conceptualizes the authors' findings and suggests that loss of sustained rhythmicity in cells with eliminated β-TrCP-mediated PER2 degradation is due to excessive nuclear repression, a prediction they verified experimentally.

Keywords

β-TrCP PERIOD2 protein degradation F-box protein CKIε mathematical modeling

References

Akashi M. , Tsuchiya Y. , Yoshino T. , and Nishida E. (2002) Control of intracellular dynamics of mammalian period proteins by casein kinase I epsilon (CKIepsilon) and CKIdelta in cultured cells . Mol Cell Biol 22: 1693-1703 .

Albrecht U. and Eichele G. (2003) The mammalian circadian clock . Curr Opin Genet Dev 13: 271-277 .

Balsalobre A. , Damiola F. , and Schibler U. (1998) A serum shock induces circadian gene expression in mammalian tissue culture cells . Cell 93: 929-937 .

Becker-Weimann S. , Wolf J. , Herzel H. , and Kramer A. (2004) Modeling feedback loops of the Mammalian circadian oscillator . Biophys J 87: 3023-3034 .

Brown SA , Fleury-Olela F. , Nagoshi E. , Hauser C. , Juge C. , Meier CA , Chicheportiche R. , Dayer JM , Albrecht U. , and Schibler U. (2005) The period length of fibroblast circadian gene expression varies widely among human individuals . PLoS Biol 3: e338 .

Coadou G. , Gharbi-Benarous J. , Megy S. , Bertho G. , Evrard-Todeschi N. , Segeral E. , Benarous R. , and Girault JP (2003) NMR studies of the phosphorylation motif of the HIV-1 protein Vpu bound to the F-box protein beta-TrCP . Biochemistry 42: 14741-14751 .

Doedel EJ (1981) AUTO: a program for the automatic bifurcation analysis of autonomous systems . Congressus Numerantium 30: 265-284 .

Eide EJ , Vielhaber EL , Hinz WA , and Virshup DM (2002) The circadian regulatory proteins BMAL1 and cryptochromes are substrates of casein kinase Iepsilon . J Biol Chem 277: 17248-17254 .

Eide EJ , Woolf MF , Kang H. , Woolf P. , Hurst W. , Camacho F. , Vielhaber EL , Giovanni A. , and Virshup DM (2005) Control of mammalian circadian rhythm by CKIepsilon-regulated proteasome-mediated PER2 degradation . Mol Cell Biol 25: 2795-2807 .

10.

Goodwin BC (1965) Oscillatory behavior in enzymatic control processes . Adv Enzyme Regul 3: 425-438 .

11.

Griffin EA Jr , Staknis D. , and Weitz CJ (1999) Light-independent role of CRY1 and CRY2 in the mammalian circadian clock . Science 286: 768-771 .

12.

Grima B. , Lamouroux A. , Chelot E. , Papin C. , Limbourg-Bouchon B. , and Rouyer F. (2002) The F-box protein slimb controls the levels of clock proteins period and timeless . Nature 420: 178-182 .

13.

Harms E. , Kivimae S. , Young MW , and Saez L. (2004) Posttranscriptional and posttranslational regulation of clock genes . J Biol Rhythms 19: 361-373 .

14.

He Q. , Cheng P. , Yang Y. , He Q. , Yu H. , and Liu Y. (2003) FWD1-mediated degradation of FREQUENCY in Neurospora establishes a conserved mechanism for circadian clock regulation . EMBO J 22: 4421-4430 .

15.

He Q. and Liu Y. (2005) Degradation of the Neurospora circadian clock protein FREQUENCY through the ubiquitin-proteasome pathway . Biochem Soc Trans 33: 953-956 .

16.

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 .

17.

Kume K. , Zylka MJ , Sriram S. , Shearman LP , Weaver DR , Jin X. , Maywood ES , Hastings MH , and Reppert SM (1999) mCRY1 and mCRY2 are essential components of the negative limb of the circadian clock feedback loop . Cell 98: 193-205 .

18.

Lee C. , Etchegaray JP , Cagampang FR , Loudon AS , and Reppert SM (2001) Posttranslational mechanisms regulate the mammalian circadian clock . Cell 107: 855-867 .

19.

Leloup JC and Goldbeter A. (2004) Modeling the mammalian circadian clock: sensitivity analysis and multiplicity of oscillatory mechanisms . J Theor Biol 230: 541-562 .

20.

Merrow M. , Spoelstra K. , and Roenneberg T. (2005) The circadian cycle: daily rhythms from behaviour to genes . EMBO Rep 6: 930-935 .

21.

Nagoshi E. , Saini C. , Bauer C. , Laroche T. , Naef F. , and Schibler U. (2004) Circadian gene expression in individual fibroblasts: cell-autonomous and self-sustained oscillators pass time to daughter cells . Cell 119: 693-705 .

22.

O'Gorman S. , Fox DT , and Wahl GM ( 1991) Recombinasemediated gene activation and site-specific integration in mammalian cells . Science 251: 1351-1355 .

23.

Pando MP , Morse D. , Cermakian N. , and Sassone-Corsi P. (2002) Phenotypic rescue of a peripheral clock genetic defect via SCN hierarchical dominance . Cell 110: 107-117 .

24.

Price JL , Blau J. , Rothenfluh A. , Abodeely M. , Kloss B. , and Young MW (1998) double-time is a novel Drosophila clock gene that regulates PERIOD protein accumulation . Cell 94: 83-95 .

25.

Rena G. , Bain J. , Elliott M. , and Cohen P. (2004) D4476, a cellpermeant inhibitor of CK1, suppresses the site-specific phosphorylation and nuclear exclusion of FOXO1a . EMBO Rep 5: 60-65 .

26.

Ripperger JA and Schibler U. (2006) Rhythmic CLOCK-BMAL1 binding to multiple E-box motifs drives circadian Dbp transcription and chromatin transitions . Nat Genet 38: 369-374 .

27.

Schlosser A. , Vanselow JT , and Kramer A. (2005) Mapping of phosphorylation sites by a multi-protease approach with specific phosphopeptide enrichment and NanoLC-MS/MS analysis . Anal Chem 77: 5243-5250 .

28.

Shearman LP , Sriram S. , Weaver DR , Maywood ES , Chaves I. , Zheng B. , Kume K. , Lee CC , van der Horst GT , Hastings MH , et al (2000) Interacting molecular loops in the mammalian circadian clock . Science 288: 1013-1019 .

29.

Shirogane T. , Jin J. , Ang XL , and Harper JW (2005) SCFbeta-TRCP controls clock-dependent transcription via casein kinase 1-dependent degradation of the mammalian period-1 (Per1) protein . J Biol Chem 280: 26863-26872 .

30.

Toh KL , Jones CR , He Y. , Eide EJ , Hinz WA , Virshup DM , Ptacek LJ , and Fu YH (2001) An hPer2 phosphorylation site mutation in familial advanced sleep phase syndrome . Science 291: 1040-1043 .

31.

Ueda HR , Hayashi S. , Chen W. , Sano M. , Machida M. , Shigeyoshi Y. , Iino M. , and Hashimoto S. (2005) System-level identification of transcriptional circuits underlying mammalian circadian clocks . Nat Genet 37: 187-192 .

32.

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 .

33.

Vielhaber E. , Eide E. , Rivers A. , Gao ZH , and Virshup DM (2000) Nuclear entry of the circadian regulator mPER1 is controlled by mammalian casein kinase I epsilon . Mol Cell Biol 20: 4888-4899 .

34.

Watanabe N. , Arai H. , Nishihara Y. , Taniguchi M. , Watanabe N. , Hunter T. , and Osada H. (2004) M-phase kinases induce phospho-dependent ubiquitination of somatic Wee1 by SCFbeta-TrCP . Proc Natl Acad Sci U S A 101: 4419-4424 .

35.

Wolf J. , Becker-Weimann S. , and Heinrich R. (2005) Analysing the robustness of cellular rhythms . Syst Biol (Stevenage) 2: 35-41 .

36.

Wu G. , Xu G. , Schulman BA , Jeffrey PD , Harper JW , and Pavletich NP (2003) Structure of a beta-TrCP1-Skp1-beta-catenin complex: destruction motif binding and lysine specificity of the SCF(beta-TrCP1) ubiquitin ligase . Mol Cell 11: 1445-1456 .

37.

Yagita K. , Tamanini F. , Der Horst GT , and Okamura H. (2001) Molecular mechanisms of the biological clock in cultured fibroblasts . Science 292: 278-281 .

38.

Zhao WN , Malinin N. , Yang FC , Staknis D. , Gekakis N. , Maier B. , Reischl S. , Kramer A. , and Weitz CJ (2007) CIPC is a mammalian circadian clock protein without invertebrate homologues . Nat Cell Biol 9: 268-275 .