Nucleocytoplasmic shuttling and mCRY-dependent inhibition of ubiquitylation of the mPER2 clock protein.
The core oscillator generating circadian rhythms in eukaryotes is composed of transcription--translation-based autoregulatory feedback loops in which clock gene products negatively affect their own expression. A key step in this mechanism involves the periodic nuclear accumulation of clock proteins following their mRNA rhythms after approximately 6 h delay. Nuclear accumulation of mPER2 is promoted by mCRY proteins. Here, using COS7 cells and mCry1/mCry2 double mutant mouse embryonic fibroblasts transiently expressing GFP-tagged (mutant) mPER2, we show that the protein shuttles between nucleus and cytoplasm using functional nuclear localization and nuclear export sequences. Moreover, we provide evidence that mCRY proteins prevent ubiquitylation of mPER2 and subsequent degradation of the latter protein by the proteasome system. Interestingly, mPER2 in turn prevents ubiquitylation and degradation of mCRY proteins. On the basis of these data we propose a model in which shuttling mPER2 is ubiquitylated and degraded by the proteasome unless it is retained in the nucleus by mCRY proteins.
|Keywords||*Biological Clocks, *Drosophila Proteins, *Eye Proteins, *Photoreceptors, Invertebrate, 3T3 Cells, Active Transport, Cell Nucleus, Animals, COS Cells, Cell Nucleus/metabolism, Cercopithecus aethiops, Cytoplasm/metabolism, Flavoproteins/genetics/*metabolism, Hela Cells, Humans, Mice, Nuclear Proteins/genetics/*metabolism, Nucleocytoplasmic Transport Proteins/genetics/*metabolism, Protein Sorting Signals, Recombinant Fusion Proteins/genetics/metabolism, Research Support, Non-U.S. Gov't, Transcription Factors, Ubiquitin/*metabolism|
|Persistent URL||dx.doi.org/10.1093/emboj/21.6.1301, hdl.handle.net/1765/13054|
Yagita, K., Tamanini, F., Yasuda, M., Hoeijmakers, J.H.J., van der Horst, G.T.J., & Okamura, H.. (2002). Nucleocytoplasmic shuttling and mCRY-dependent inhibition of ubiquitylation of the mPER2 clock protein.. EMBO Journal, 21(6), 1301–1314. doi:10.1093/emboj/21.6.1301