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Control of Mammalian Circadian Rhythm by CKIε-Regulated Proteasome-Mediated PER2 Degradation
Open accessAbstract
The mammalian circadian regulatory proteins PER1 and PER2 undergo a daily cycle of accumulation followed by phosphorylation and degradation. Although phosphorylation-regulated proteolysis of these inhibitors is postulated to be essential for the function of the clock, inhibition of this process has not yet been shown to alter mammalian circadian rhythm. We have developed a cell-based model of PER2 degradation. Murine PER2 (mPER2) hyperphosphorylation induced by the cell-permeable protein phosphatase inhibitor calyculin A is rapidly followed by ubiquitination and degradation by the 26S proteasome. Proteasome-mediated degradation is critically important in the circadian clock, as proteasome inhibitors cause a significant lengthening of the circadian period in Rat-1 cells. CKIepsilon (casein kinase Iepsilon) has been postulated to prime PER2 for degradation. Supporting this idea, CKIepsilon inhibition also causes a significant lengthening of circadian period in synchronized Rat-1 cells. CKIepsilon inhibition also slows the degradation of PER2 in cells. CKIepsilon-mediated phosphorylation of PER2 recruits the ubiquitin ligase adapter protein beta-TrCP to a specific site, and dominant negative beta-TrCP blocks phosphorylation-dependent degradation of mPER2. These results provide a biochemical mechanism and functional relevance for the observed phosphorylation-degradation cycle of mammalian PER2. Cell culture-based biochemical assays combined with measurement of cell-based rhythm complement genetic studies to elucidate basic mechanisms controlling the mammalian clock.
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Metadata
- Authors
- Eide, E. J., Woolf, M. F., Kang, H., Woolf, P., Hurst, W., Camacho, F., Vielhaber, E. L., Giovanni, A., & Virshup, D. M.
- Affiliations
- Huntsman Cancer Institute (US)
- University of Michigan (US)
- Aventis Pharmaceuticals, Bridgewater, New Jersey
- University of Utah (US)
- Corresponding author
- Virshup, David M.
- Venue
- Molecular and Cellular Biology
- Publisher
- Informa UK Limited
- Published
- 2005-04-01
- Volume
- 25
- Issue
- 7
- Pages
- 2795-2807
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- PMC
- PMC1061645
- OpenAlex
- W1990361088
- Cited by
- 495
- References
- 54
- Citations by year
- 2026: 10 · 2025: 24 · 2024: 14 · 2023: 16 · 2022: 26 · 2021: 22 · 2020: 33 · 2019: 25 · 2018: 21 · 2017: 22 · 2016: 23 · 2015: 24 · 2014: 24 · 2013: 23 · 2012: 20
- FWCI
- 10.27
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Molecular and Cellular Biology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Huntsman Cancer Foundation
- National Institutes of Health
- National Cancer Institute (P04CA42104)
- National Institute of General Medical Sciences (R01 GM60387, R01GM060387)
- Topics
- Life Sciences › Neuroscience › Endocrine and Autonomic Systems › Circadian rhythm and melatonin
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Light effects on plants
- Keywords
- circadian clock, casein kinase I epsilon, proteasome-mediated proteolysis, ubiquitination, calyculin A, Rat-1 cells
- MeSH terms
- Animals, Cell Line, Circadian Rhythm, Gene Expression Regulation, Humans, Nuclear Proteins, Phosphorylation, Protein Binding, Transcription Factors, Cell Cycle Proteins, Ubiquitin-Protein Ligases, beta-Transducin Repeat-Containing Proteins, Proteasome Endopeptidase Complex, Casein Kinase 1 epsilon, Mice, Rats, Period Circadian Proteins
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