Abstract
Cyclin D1 regulates G1 progression. Its transcriptional regulation is well understood. However, the mechanism underlying cyclin D1 ubiquitination and its subsequent degradation is not yet clear. We report that cyclin D1 undergoes increased degradation in the cytoplasm during S phase in a variety of cancer cells. This is mediated by phosphorylation at Thr286 through the activity of the Ras/Raf/MEK/ERK cascade and the F-box protein FBXW8, which is an E3 ligase. The majority of FBXW8 is expressed in the cytoplasm during G1 and S phase. In contrast, cyclin D1 accumulates in the nucleus during G1 phase and exits into the cytoplasm in S phase. Increased cyclin D1 degradation is linked to association with FBXW8 in the cytoplasm, and enhanced phosphorylation of cyclin D1 through sustained ERK1/2 signaling. Depletion of FBXW8 caused a significant accumulation of cyclin D1, as well as sequestration of CDK1 in the cytoplasm. This resulted in a severe reduction of cell proliferation. These effects could be rescued by constitutive nuclear expression of cyclin D1-T286A. Thus, FBXW8 plays an essential role in cancer cell proliferation through proteolysis of cyclin D1. It may present new opportunities to develop therapies targeting destruction of cyclin D1 or its regulator E3 ligase selectively.
MeSH Terms
Amino Acid Sequence
Amino Acid Substitution
Animals
Base Sequence
Binding Sites
Cell Cycle
Cell Line, Tumor
Cell Proliferation
Cyclin D1/chemistry,genetics,metabolism
F-Box Proteins/antagonists & inhibitors,genetics,metabolism
Humans
In Vitro Techniques
MAP Kinase Signaling System
Mice
Multiprotein Complexes
Mutagenesis, Site-Directed
NIH 3T3 Cells
Proteasome Endopeptidase Complex/metabolism
Protein Stability
Protein Structure, Tertiary
RNA, Small Interfering/genetics
Recombinant Fusion Proteins/chemistry,genetics,metabolism
Threonine/chemistry
Ubiquitin/metabolism
Chemicals
CCND1 protein, human
Ccnd1 protein, mouse
F-Box Proteins
FBXW8 protein, human
Fbxw8 protein, mouse
Multiprotein Complexes
RNA, Small Interfering
Recombinant Fusion Proteins
Ubiquitin
Cyclin D1
Threonine
Proteasome Endopeptidase Complex
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Okabe Hiroshi
Department of Pathology, School of Medicine, University of California San Francisco, San Francisco, California, United States of America; UCSF Comprehensive Cancer Center, School of Medicine, University of California San Francisco, San Francisco, California, United States of America.
Lee Sang-Hyun
Phuchareon Janyaporn
Albertson Donna G
McCormick Frank
Tetsu Osamu
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