Zhang Mengchen, Yao Feng, Qin Tong, Hou Lin, Zou Xiangyang
College of Life Sciences, Liaoning Normal University, Dalian 116081, China.
College of Life Sciences, Liaoning Normal University, Dalian 116081, China.
Gene. 2017 Apr 15;608:28-40. doi: 10.1016/j.gene.2017.01.011. Epub 2017 Jan 21.
Proper control of the cellular processes requires a variety of regulatory proteins that are involved in the cell cycle, proliferation and apoptosis. Cyclin-dependent kinase inhibitor (CKI) negatively regulates transcription and arrests the cell cycle in G phase. KIP2 is a member of CKI family, which could inhibit proliferation by tight-binding with several cyclin-CDK complexes. During the embryonic development of the brine shrimp, Artemia sinica, KIP2 plays a key role in the cell cycle regulation, but the specific mechanisms remain unknown. Herein, the 1023bp full-length cDNA of kip2 from A. sinica was cloned. The mRNA expression patterns of As-kip2, As-carp-1 in different development stages and pattern of As-kip2 under environmental stresses were investigated. In situ hybridization of As-kip2 mRNA and immunofluorescence of As-CARP-1 protein showed no tissue or organ specificity. Furthermore, western blotting showed the expressions levels of As-KIP2, As-E2F1, As-p53, As-cyclin E, As-SODD protein, and pattern of As-KIP2 under environmental stresses. Our research revealed that As-KIP2 plays crucial role in the restarting process of diapause embryo in Artemia sinica.
细胞过程的适当调控需要多种参与细胞周期、增殖和凋亡的调节蛋白。细胞周期蛋白依赖性激酶抑制剂（CKI）对转录起负调控作用，并使细胞周期停滞在G期。KIP2是CKI家族的成员之一，它可以通过与几种细胞周期蛋白-CDK复合物紧密结合来抑制增殖。在卤虫中华卤虫的胚胎发育过程中，KIP2在细胞周期调控中起关键作用，但具体机制尚不清楚。在此，克隆了中华卤虫kip2的1023bp全长cDNA。研究了As-kip2、As-carp-1在不同发育阶段的mRNA表达模式以及As-kip2在环境胁迫下的表达模式。As-kip2 mRNA的原位杂交和As-CARP-1蛋白的免疫荧光显示没有组织或器官特异性。此外，蛋白质印迹法显示了As-KIP2、As-E2F1、As-p53、As-细胞周期蛋白E、As-SODD蛋白的表达水平以及As-KIP2在环境胁迫下的表达模式。我们的研究表明，As-KIP2在中华卤虫滞育胚胎的重启过程中起关键作用。
