Goubin F, Ducommun B
Laboratoire de Pharmacologie et de Toxicologie Fondamentales, CNRS, Université Paul Sabatier, Toulouse III, France.
Oncogene. 1995 Jun 15;10(12):2281-7.
Members of the recently discovered family of cyclin-dependent kinases inhibitors (CKIs) appear to play an essential regulatory role in the control of cell proliferation. To investigate the molecular basis of the interaction between these proteins and the cyclin-dependent kinases (CDKs), we performed a systematic mutagenesis of the CKI family member p21Cip1 using the alanine-scanning strategy. We have examined the interaction between in vitro translated human cdk2, cyclins A and D1, purified proliferating cell nuclear antigen (PCNA) and a set of human p21Cip1 mutants fused to glutathione S-transferase. Independent domains that are required for the interaction with cdk2 and with PCNA have been identified. The cdk2 binding domain is located in the N-terminal part of the protein, between residues 45 and 60, a region that is fully conserved in the p27Kip1 inhibitor. A PCNA binding region was localised to the C-terminus of the protein, between residues 142 and 163. These findings define protein motifs that are highly conserved between members of the CKI family and that are likely to play an essential function in the regulation of the G1/S transition.
最近发现的细胞周期蛋白依赖性激酶抑制剂（CKIs）家族成员似乎在细胞增殖控制中发挥着重要的调节作用。为了研究这些蛋白质与细胞周期蛋白依赖性激酶（CDKs）之间相互作用的分子基础，我们使用丙氨酸扫描策略对CKI家族成员p21Cip1进行了系统诱变。我们检测了体外翻译的人cdk2、细胞周期蛋白A和D1、纯化的增殖细胞核抗原（PCNA）与一组与谷胱甘肽S-转移酶融合的人p21Cip1突变体之间的相互作用。已经确定了与cdk2和PCNA相互作用所需的独立结构域。cdk2结合结构域位于蛋白质的N端部分，在第45至60位残基之间，该区域在p27Kip1抑制剂中完全保守。PCNA结合区域定位于蛋白质的C端，在第142至163位残基之间。这些发现定义了CKI家族成员之间高度保守的蛋白质基序，这些基序可能在G1/S期转换的调节中发挥重要作用。
