1. Institut de Neurobiologie Alfred Fessard (NGI, CNRS UPR 2216) and Centre de Génétique Moléculaire (CNRS UPR 2167), Centre National de la Recherche Scientifique, av. de la terrasse, 91198 Gif-sur-Yvette, France

Correspondence to: François Rouyer¹ Correspondence and requests for materials should be addressed to F.R. (e-mail: Email: rouyer@iaf.cnrs-gif.fr).

Abstract

The Drosophila circadian clock is driven by daily fluctuations of the proteins Period and Timeless, which associate in a complex and negatively regulate the transcription of their own genes^{1, 2}. Protein phosphorylation has a central role in this feedback loop, by controlling Per stability in both cytoplasmic and nuclear compartments^{3, 4, 5, 6} as well as Per/Tim nuclear transfer^{7, 8}. However, the pathways regulating degradation of phosphorylated Per and Tim are unknown. Here we show that the product of the slimb (slmb) gene⁹—a member of the F-box/WD40 protein family of the ubiquitin ligase SCF complex that targets phosphorylated proteins for degradation^{10, 11, 12, 13}—is an essential component of the Drosophila circadian clock. slmb mutants are behaviourally arrhythmic, and can be rescued by targeted expression of Slmb in the clock neurons. In constant darkness, highly phosphorylated forms of the Per and Tim proteins are constitutively present in the mutants, indicating that the control of their cyclic degradation is impaired. Because levels of Per and Tim oscillate in slmb mutants maintained in light:dark conditions, light- and clock-controlled degradation of Per and Tim do not rely on the same mechanisms.