The role of {beta}-TrCP1 and {beta}-TrCP2 in circadian rhythm generation by mediating degradation of clock protein PER2.
The mammalian circadian clock proteins undergo a daily cycle of accumulation followed by phosphorylation and degradation. The mechanism by which clock proteins undergo degradation has not been fully understood. Circadian clock protein PERIOD2 (PER2) is shown to be the potential target of F-box protein beta-TrCP1, a component of ubiquitin E3 ligase. Here, we show that beta-TrCP2 as well as beta-TrCP1 target PER2 protein in vitro. We also identified beta-TrCP binding site (m2) of PER2 being recognized by both beta-TrCP1 and beta-TrCP2. Luciferase-PER2 fusion system revealed that m2 site was responsible for the stability of PER2. The role of beta-TrCP1 and beta-TrCP2 in circadian rhythm generation was analysed by real-time reporter assay revealing that siRNA-mediated suppressions of beta-TrCP1 and/or beta-TrCP2 attenuate circadian oscillations in NIH3T3 cell. beta-TrCP1-deficient mice, however, showed normal period length, light-induced phase-shift response in behaviour and normal expression of PER2, suggesting that beta-TrCP1 is dispensable for the central clock in the suprachiasmatic nucleus. Our study indicates that beta-TrCP1 and beta-TrCP2 were involved in the cell autonomous circadian rhythm generation in culture cells, although the role of beta-TrCP2 in the central clock in the suprachiasmatic nucleus remains to be elucidated.
Ohsaki
Kanae
K
Clock Cell Biology Research Group, Advanced Industrial Science and Technology, Ibaraki, Japan.
Oishi
Katsutaka
K
Kozono
Yuko
Y
Nakayama
Keiko
K
Nakayama
Keiichi I
KI
Ishida
Norio
N
D000818:Animals; D001683:Biological Clocks / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002940:Circadian Rhythm / Q000502:physiology*; D017930:Genes, Reporter; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D009043:Motor Activity / Q000502:physiology; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000378:metabolism*; D056950:Period Circadian Proteins; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D044844:beta-Transducin Repeat-Containing Proteins / Q000235:genetics / Q000378:metabolism*
D018797:Cell Cycle Proteins; C486774:Fbxw1b protein, mouse; D009687:Nuclear Proteins; C513312:Per2 protein, mouse; C513311:Per2 protein, rat; D056950:Period Circadian Proteins; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D014157:Transcription Factors; D044844:beta-Transducin Repeat-Containing Proteins