Interaction of the CDK2-associated protein-1, p12DOC-1/CDK2AP1, with its homolog, p14DOC-1R
Interaction of the CDK2-associated protein-1, p12DOC-1/CDK2AP1, with its homolog, p14DOC-1R
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DOI：
10.1016/j.bbrc.2004.02.003
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发表时间：
2004-03-19
影响因子：
3.1
通讯作者：
Wong, DTW
中科院分区：
文献类型：
--
作者：
Buajeeb, W;Zhang, X;Wong, DTW
人DOC-1/CDK 2AP 1基因编码12 kDa的生长抑制蛋白（p12（DOC-1/CDK 2AP 1））。最近，p12（DOC-1/CDK 2AP 1）已被证明与细胞周期蛋白，包括CDK 2和DNA聚合酶α/引物。它负调节CDK 2活性并抑制DNA复制。因此，其他p12（DOC-1/CDK 2AP 1）相互作用蛋白的鉴定可能阐明其在细胞周期调控和癌变中的作用。本研究的目的是利用酵母双杂交系统鉴定p12（DOC-1/CDK 2AP 1）相互作用蛋白。以人p12（DOC-1/CDK 2AP 1）为诱饵，从肝脏cDNA文库中筛选出与人DOC-1 R转录本相同的cDNA克隆。p12（DOC-1/CDK 2AP 1）和p14（DOC-1 R）之间的相互作用在体外和细胞中得到验证。GST pull-down实验和免疫沉淀实验证实了两种蛋白之间的相互作用。在酵母双杂交系统中，利用一系列缺失突变体作为诱饵，将p12（DOC-1/CDK 2AP 1）与p14（DOC-1 R）相互作用的关键区域确定为20-25位氨基酸。p12（DOC-1/CDK 2AP 1）与其同源蛋白p14（DOC-1 R）结合。(C)2004年爱思唯尔公司All rights reserved.
Human DOC-1/CDK2AP1 gene encodes a growth suppressor protein of 12 kDa (p12(DOC-1/CDK2AP1)). Recently, p12(DOC-1/CDK2AP1) has been shown to associate with cell cycle proteins including CDK2 and DNA polymerase alpha/primase. It negatively regulates CDK2 activities and suppresses DNA replication. Therefore, identification of other p12(DOC-1/CDK2AP1) interacting proteins might clarify its role in the cell cycle regulation and carcinogenesis. The purpose of this study was to identify additional p12(DOC-1/CDK2AP1) interacting proteins using the yeast two-hybrid system. Using human p12(DOC-1/CDK2AP1) as a bait in a liver cDNA library screening, cDNA clones identical to human DOC-1R transcript were identified. The interaction between p12(DOC-1/CDK2AP1) and p14(DOC-1R) was verified in vitro and in cells. GST pull-down assay and immunoprecipitation experiments confirmed the interaction between the two proteins. The critical region for p12(DOC-1/CDK2AP1)'s interaction with p14(DOC-1R) was defined to amino acids 20-25 by using a series of deletion mutants as baits in the yeast two-hybrid system. Our data indicated that p12(DOC-1/CDK2AP1) could associate with its homologous protein, p14(DOC-1R). (C) 2004 Elsevier Inc. All rights reserved.