Madsen P, Celis J E
FEBS Lett. 1985 Nov 25;193(1):5-11. doi: 10.1016/0014-5793(85)80068-5.
The sequence of cyclin (proliferating cell nuclear antigen, PCNA), antigen staining throughout the cell cycle of African green monkey kidney cells (BS-C-1) has been determined by indirect immunofluorescence using PCNA autoantibodies specific for this protein. Patterns of cyclin staining observed between the beginning of S-phase and maximum DNA synthesis are similar to those reported in human AMA cells [(1985) Proc. Natl. Acad. Sci. USA 82, 3262-3266], while those detected thereafter are significantly different; the most striking feature being the continuous staining of the nucleoli up to or very near the S/G2 border of the cell cycle. Using [3H]thymidine autoradiography and indirect immunofluorescence of the same cells we show a remarkable correlation between cyclin antigen distribution and topographical patterns of DNA synthesis. In addition, we present evidence showing that DNase I treatment of Triton-extracted monolayers abolishes cyclin antigen staining but does not result in a substantial release of this protein. Taken together the above observations argue for a role of cyclin in some aspect of DNA replication.
利用针对该蛋白的增殖细胞核抗原(PCNA)自身抗体,通过间接免疫荧光法确定了非洲绿猴肾细胞(BS-C-1)整个细胞周期中细胞周期蛋白(增殖细胞核抗原,PCNA)的抗原染色序列。在S期开始至DNA合成达到最大值之间观察到的细胞周期蛋白染色模式与人类AMA细胞中报道的模式相似[(1985年)《美国国家科学院院刊》82卷,3262 - 3266页],而此后检测到的模式则有显著差异;最显著的特征是核仁持续染色直至细胞周期的S/G2边界或非常接近该边界。使用[3H]胸腺嘧啶核苷放射自显影术和对相同细胞的间接免疫荧光法,我们展示了细胞周期蛋白抗原分布与DNA合成的拓扑模式之间存在显著相关性。此外,我们提供的证据表明,用脱氧核糖核酸酶I处理经曲拉通提取的单层细胞可消除细胞周期蛋白抗原染色,但不会导致该蛋白大量释放。综合上述观察结果,表明细胞周期蛋白在DNA复制的某些方面发挥作用。