Abstract
Protein phosphorylation has a key role in modulating the stabilities of circadian clock proteins in a manner specific to the time of day1. A conserved feature of animal clocks is that Period (Per) proteins undergo daily rhythms in phosphorylation and levels2,3, events that are crucial for normal clock progression4,5,6,7. Casein kinase Iε (CKIε) has a prominent role in regulating the phosphorylation and abundance of Per proteins in animals8. This was first shown in Drosophila with the characterization of Doubletime (Dbt), a homologue of vertebrate casein kinase Iε4,6. However, it is not clear how Dbt regulates the levels of Per. Here we show, using a cell culture system, that Dbt promotes the progressive phosphorylation of Per, leading to the rapid degradation of hyperphosphorylated isoforms by the ubiquitin–proteasome pathway. Slimb, an F-box/WD40-repeat protein functioning in the ubiquitin–proteasome pathway9,10 interacts preferentially with phosphorylated Per and stimulates its degradation. Overexpression of slimb or expression in clock cells of a dominant-negative version of slimb disrupts normal rhythmic activity in flies. Our findings suggest that hyperphosphorylated Per is targeted to the proteasome by interactions with Slimb.
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Acknowledgements
We thank S. Kay for the pAct-per and pAct-tim constructs, A. Seghal for the pCasper-hs-Ub/HA construct, J. Hall for the tim-gal4-27 and tim-gal4-62 lines, and M. Young for the tim-(UAS)-gal4 line. The work was supported by the Searle Scholar Program and the UTSW Endowed Scholar Program to J.J., and by a grant from the National Institutes of Health to I.E.
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Ko, H., Jiang, J. & Edery, I. Role for Slimb in the degradation of Drosophila Period protein phosphorylated by Doubletime. Nature 420, 673–678 (2002). https://doi.org/10.1038/nature01272
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DOI: https://doi.org/10.1038/nature01272
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