Castro A, Jaumot M, Vergés M, Agell N, Bachs O
Departament de Biologia Cellular, Facultat de Medicina, Universitat de Barcelona, Spain.
Biochem Biophys Res Commun. 1994 Jun 30;201(3):1072-8. doi: 10.1006/bbrc.1994.1814.
The expression and intracellular localization of cyclin A and cdk2 have been analyzed in rat liver cells proliferatively activated in vivo by a partial hepatectomy. Western blot analysis revealed that cyclin A started to increase during G1 (at 6 h after hepatectomy) reaching maximal levels during S phase (at 18 h). Cdk2 began to increase during late G1 (at 12 h) peaking also at 18 h. At the latter time cyclin A was mainly localized in the microsomal fraction, although it was also present in cytosol, plasma membrane and nucleus. Active cyclin/cdks complexes containing cyclin A and cdk2 were obtained by precipitation with p13-Sepharose after solubilization of microsomes with triton X-100. The presence of active cyclin A/cdk2 complexes in microsomes was confirmed by immunoprecipitation experiments with anti-cdk2 antibodies. These results suggest a putative role of cyclin A/cdk2 during S phase which would be related with microsomal function.
通过部分肝切除术在体内增殖激活的大鼠肝细胞中,对细胞周期蛋白A(cyclin A)和细胞周期蛋白依赖性激酶2(cdk2)的表达及细胞内定位进行了分析。蛋白质免疫印迹分析显示,细胞周期蛋白A在G1期(肝切除术后6小时)开始增加,在S期(18小时)达到最高水平。cdk2在G1期末期(12小时)开始增加,同样在18小时达到峰值。在18小时时,细胞周期蛋白A主要定位于微粒体部分,不过在胞质溶胶、质膜和细胞核中也有存在。在用曲拉通X-100溶解微粒体后,通过p13-琼脂糖凝胶沉淀获得了含有细胞周期蛋白A和cdk2的活性细胞周期蛋白/细胞周期蛋白依赖性激酶复合物。用抗cdk2抗体进行免疫沉淀实验证实了微粒体中存在活性细胞周期蛋白A/cdk2复合物。这些结果表明细胞周期蛋白A/cdk2在S期具有假定作用,这可能与微粒体功能有关。