Kimura Masashi, Uchida Chiharu, Takano Yukihiko, Kitagawa Masatoshi, Okano Yukio
Department of Molecular Pathobiochemistry, Gifu University School of Medicine, Tsukasamachi-40, Gifu 500-8705, Japan.
Biochem Biophys Res Commun. 2004 Apr 9;316(3):930-6. doi: 10.1016/j.bbrc.2004.01.178.
Aurora B is an important regulator of mitosis, and its mRNA and protein levels are tightly regulated during the cell cycle. In this study, we cloned the 5' flanking region of the human aurora B gene and characterized its promoter activity. Two major transcription initiation sites were identified by primer extension. aurora B promoter activity was upregulated during M phase, and its cell cycle-dependent element (CDE) and cell cycle-gene homology region (CHR) upstream of the transcription initiation sites regulated the cell cycle-dependent promoter activity. Several CDE-binding protein complexes were identified using the electrophoretic mobility shift assay. Using the biotin-streptavidin pull-down assay, binding of E2F-1, E2F-4, and DP-2, but not of DP-1, to the CDE was detected. These results demonstrate that aurora B mRNA level is regulated by CDE-CHR and that a subset of E2F/DP family proteins binds to the CDE.
极光激酶B是有丝分裂的重要调节因子,其mRNA和蛋白质水平在细胞周期中受到严格调控。在本研究中,我们克隆了人类极光激酶B基因的5'侧翼区域,并对其启动子活性进行了表征。通过引物延伸鉴定出两个主要转录起始位点。极光激酶B启动子活性在M期上调,其转录起始位点上游的细胞周期依赖性元件(CDE)和细胞周期基因同源区域(CHR)调节细胞周期依赖性启动子活性。使用电泳迁移率变动分析鉴定了几种CDE结合蛋白复合物。使用生物素-链霉亲和素下拉分析,检测到E2F-1、E2F-4和DP-2与CDE结合,但未检测到DP-1与CDE结合。这些结果表明,极光激酶B mRNA水平受CDE-CHR调控,并且E2F/DP家族蛋白的一个子集与CDE结合。