WAF1 retards S-phase progression primarily by inhibition of cyclin-dependent kinases
WAF1 retards S-phase progression primarily by inhibition of cyclin-dependent kinases
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DOI：
10.1128/mcb.17.8.4877
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发表时间：
1997-08-01
影响因子：
5.3
通讯作者：
Howard, BH
中科院分区：
文献类型：
--
作者：
Ogryzko, VV;Wong, P;Howard, BH
p(2WAF1/CIP1/sdi1)基因产物(WAF1)在体外抑制DNA复制(J. Chen, P. Jackson, M. Kirschner, and A. Dutta, Nature 374:386-388, 1995; S. Waga, G. Hannon, D. Beach, and B. Stillman, Nature 369:574-578, 1994)，但体内研究WAF1 的抗增殖活性尚未解决因 S 期进展的潜在抑制而导致的 G(1) 期停滞。在这里，我们证明 WAF1 表达升高可以阻碍体内复制 DNA 合成。 WAF1介导的抑制作用可以被细胞周期蛋白A、细胞周期蛋白E或猿猴病毒40小t抗原拮抗，而转染细胞中WAF1蛋白水平没有降低，增殖细胞核抗原（PCNA）过表达对于拮抗WAF1作用既不是必要的也不是充分的，负责细胞周期蛋白依赖性激酶（CDK）相互作用的WAF1 N末端结构域的表达与全长 WAF1，而 PCNA 结合 C 末端表现出适度的活性。我们得出的结论是，哺乳动物细胞中的 S 期进展依赖于持续的细胞周期蛋白和 CDK 活性，并且 WAF1 主要通过细胞周期蛋白和 CDK 依赖性途径影响 S 期。
The p(2WAF1/CIP1/sdi1) gene product (WAF1) inhibits DNA replication in vitro (J. Chen, P. Jackson, M. Kirschner, and A. Dutta, Nature 374:386-388, 1995; S. Waga, G. Hannon, D. Beach, and B. Stillman, Nature 369:574-578, 1994), but in vivo studies on the antiproliferative activity of WAF1 have not resolved G(1)-phase arrest from potential inhibition of S-phase progression. Here, we demonstrate that elevated WAF1 expression can retard replicative DNA synthesis in vivo. The WAF1-mediated inhibitory effect could be antagonized by cyclin A, cyclin E, or the simian virus 40 small-t antigen with no decrease in the levels of WAF1 protein in transfected cells, Proliferating-cell nuclear antigen (PCNA) overexpression was neither necessary nor sufficient to antagonize WAF1 action, Expression of the N-terminal domain of WAF1, responsible for cyclin-dependent kinase (CDK) interaction, had the same effect as full-length WAF1, while the PCNA binding C terminus exhibited modest activity. We conclude that S-phase progression in mammalian cells is dependent on continuing cyclin and CDK activity and that WAF1 affects S phase primarily through cyclin- and CDK-dependent pathways.