Molecular Cloning and Functional Analysis of CDK Variant Expressed in Breast Cancer Tissues

我们从breast cancer tissues， and investigated its function and the possibility of similar variant in other CDK family中克隆了一个T-loop deletion variant（CDC 2）。CDC 2 ΔT-HA在带有cyclin B1和/或21 CDK抑制剂的Hela细胞中临时协同表示。蛋白质-蛋白质相互作用是由Yi D135 Yie D1 S免疫吸收（IP）或IP-西方研究的，以及免疫复合体的基因活动是由历史-H1激酶试验研究的。Although CDC2ΔT possesses the conserved cyclin binding site and the ATP binding site， it is unable to complex with either Cyclin B1 nor p21 and lacks histon H1 kinase activity。结果表明，T回路并不只是在保持免费CDK在其不活跃状态下发挥关键作用，但也在促进循环素绑定的作用下使CDK激活。通过对CDK家族中的其他变体进行额外的研究，我们克隆了CDK 2变体，在其T-环的C-末端部分中总共有34种氨基酸。（CDK 2 ΔT，#AB012305）细胞周期或钙生成中这些变异体的生物学意义在目前是不可预测的。CDC2 and CDK2 may elacidate the role of these variants。

We have cloned a T-loop deletion variant CDC2(CDC2ΔT : described as ΔCDC2 in the application form for this grant) from breast cancer tissues, and investigated its function and the possibility of similar variant in other CDK family. CDC2ΔT-HA was transiently co-expressed in Hela cells with cyclin B1 and/or 21 CDK inhibitor. The protein-protein interaction was studied by ィイD135ィエD1S-immunoprecipitation(IP) or IP-western, and kianse activity of the immunocomplex was studied by Histo-H1 kinase assay. Although CDC2ΔT possesses the conserved cyclin binding site and the ATP binding site, it is unable to complex with either Cyclin B1 nor p21 and lacks histon H1 kinase activity. The results indicate that the T-loop not only play a key role in keeping a free CDK in its inactive state but also in facilitating CDK activation by promoting cyclin binding. Through additional study to investigate other variant in CDK family, we cloned a CDK2 variant, which lacks 34 amino acids in the C-terminal portion of its T-loop. (CDK2ΔT, #AB012305) The biological significance of these variants in cell cycle or carcinogenesis is unclear at present. Further investigation of detail function of each domain of CDC2 and CDK2 may elacidate the role of these variants.