Phosphorylation of Bcl-2 protein by CDC2 kinase during G2/M phases and its role in cell cycle regulation

被引:101
作者
Furukawa, Y
Iwase, S
Kikuchi, J
Terui, Y
Nakamura, M
Yamada, H
Kano, Y
Matsuda, M
机构
[1] Jichi Med Sch, Ctr Mol Med, Div Mol Hematopoiesis, Minami Kawachi, Tochigi 3290498, Japan
[2] Jichi Med Sch, Dept Hematol, Minami Kawachi, Tochigi 3290498, Japan
[3] Jikei Univ, Sch Med, Dept Internal Med Aoto, Tokyo 1058461, Japan
[4] Jikei Univ, Sch Med, Dept Mol Genet, Inst DNA Med, Tokyo 1058461, Japan
[5] Tochigi Canc Ctr, Div Med Oncol, Utsunomiya, Tochigi 3200834, Japan
关键词
D O I
10.1074/jbc.M906893199
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Although it has been reported that Bcl-2 phosphorylation is associated with certain types of apoptosis, there is much controversy over the functional significance of and the kinases responsible for the phosphorylation. In this study, we examined whether Bcl-2 is phosphorylated by CDC2 kinase, a master regulator of G(2)/M transition in the eukaryotic cell cycle. When CDC2 was activated by okadaic acid in HL-60 cells, Bcl-2 phosphorylation was readily induced. The phosphorylation was correlated with the accumulation of cells in G(2)/M phases, but was not proportional to the level of apoptosis. Furthermore, we found that Bcl-2 was phosphorylated during G(2)/M phases of normal cell cycle. The ability of CDC2 to phosphorylate Bcl-2 was confirmed by in vitro kinase assay with a highly purified CDC2-cyclin B complex. Using synthetic peptides and mutant cell lines, we identified threonine 56, one of two consensus sites for CDC2 within the Bcl-2 sequence, as a residue phosphorylated by CDC2. Mutation at threonine 56 abrogated the cell cycle inhibitory effect of Bcl-2 without affecting anti-apoptotic function. These results suggest that two distinct functions of Bcl-2 (anti-apoptosis and cell cycle inhibition) are differentially regulated by post-translational mechanisms such as phosphorylation. CDC2-mediated phosphorylation of Bcl-2 may play some physiological roles in the negative regulatory events during mitosis.
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页码:21661 / 21667
页数:7
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