CDK4 protein is degraded by anaphase-promoting complex/cyclosome in mitosis and reaccumulates in early G1 phase to initiate a new cell cycle in HeLa cells

Huabo Chen; Xiaowei Xu; Guopeng Wang; Boyan Zhang; Gang Wang; Guangwei Xin; Junjun Liu; Qing Jiang; Hongyin Zhang; Chuanmao Zhang

doi:10.1074/jbc.M116.773226

CDK4 protein is degraded by anaphase-promoting complex/cyclosome in mitosis and reaccumulates in early G₁ phase to initiate a new cell cycle in HeLa cells

J Biol Chem. 2017 Jun 16;292(24):10131-10141. doi: 10.1074/jbc.M116.773226. Epub 2017 Apr 26.

Authors

Affiliations

¹ From the Ministry of Education Key Laboratory of Cell Proliferation and Differentiation and the State Key Laboratory of Membrane Biology, College of Life Sciences, Peking University, Beijing 100871, China.
² the Department of Biological Sciences, California State Polytechnic University, Pomona, California 91768.
³ From the Ministry of Education Key Laboratory of Cell Proliferation and Differentiation and the State Key Laboratory of Membrane Biology, College of Life Sciences, Peking University, Beijing 100871, China, [email protected].
⁴ the Cancer Research Center, Peking University Hospital, Peking University, Beijing 100871, China, and.
⁵ From the Ministry of Education Key Laboratory of Cell Proliferation and Differentiation and the State Key Laboratory of Membrane Biology, College of Life Sciences, Peking University, Beijing 100871, China, [email protected].

Abstract

CDK4 regulates G₁/S phase transition in the mammalian cell cycle by phosphorylating retinoblastoma family proteins. However, the mechanism underlying the regulation of CDK4 activity is not fully understood. Here, we show that CDK4 protein is degraded by anaphase-promoting complex/cyclosome (APC/C) during metaphase-anaphase transition in HeLa cells, whereas its main regulator, cyclin D1, remains intact but is sequestered in cytoplasm. CDK4 protein reaccumulates in the following G₁ phase and shuttles between the nucleus and the cytoplasm to facilitate the nuclear import of cyclin D1. Without CDK4, cyclin D1 cannot enter the nucleus. Point mutations that disrupt CDK4 and cyclin D1 interaction impair the nuclear import of cyclin D1 and the activity of CDK4. RNAi knockdown of CDK4 also induces cytoplasmic retention of cyclin D1 and G₀/G₁ phase arrest of the cells. Collectively, our data demonstrate that CDK4 protein is degraded in late mitosis and reaccumulates in the following G₁ phase to facilitate the nuclear import of cyclin D1 for activation of CKD4 to initiate a new cell cycle in HeLa cells.

Keywords: cell cycle; cyclin D1; cyclin-dependent kinase (CDK); protein degradation; retinoblastoma protein (pRb, RB).

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Active Transport, Cell Nucleus
Anaphase-Promoting Complex-Cyclosome / metabolism*
Animals
Cell Line
Cyclin D1 / chemistry
Cyclin D1 / genetics
Cyclin D1 / metabolism*
Cyclin-Dependent Kinase 4 / antagonists & inhibitors
Cyclin-Dependent Kinase 4 / chemistry
Cyclin-Dependent Kinase 4 / genetics
Cyclin-Dependent Kinase 4 / metabolism*
Enzyme Induction
Enzyme Stability
G1 Phase*
Green Fluorescent Proteins / chemistry
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
HeLa Cells
Humans
Mice
Mitosis*
Neoplasm Proteins / chemistry
Neoplasm Proteins / genetics
Neoplasm Proteins / metabolism
Nuclear Localization Signals / chemistry
Nuclear Localization Signals / genetics
Nuclear Localization Signals / metabolism
Peptide Fragments / antagonists & inhibitors
Peptide Fragments / chemistry
Peptide Fragments / metabolism
Point Mutation
Protein Stability
Protein Transport
Proteolysis
RNA Interference
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / metabolism

Substances

CCND1 protein, human
Neoplasm Proteins
Nuclear Localization Signals
Peptide Fragments
Recombinant Fusion Proteins
Cyclin D1
Green Fluorescent Proteins
Anaphase-Promoting Complex-Cyclosome
CDK4 protein, human
Cyclin-Dependent Kinase 4