Thwaites Michael J, Cecchini Matthew J, Dick Frederick A
London Regional Cancer Program, London, ON, Canada.
Methods Mol Biol. 2014;1170:449-61. doi: 10.1007/978-1-4939-0888-2_24.
The G1/S-phase restriction point is an important landmark in the mammalian cell division cycle. The key regulator of the G1/S transition is the retinoblastoma gene product (pRB). It prevents the transcription of genes required for S-phase progression by repressing E2F transcription factors. An increase in Cdk phosphorylation of pRB causes the release of E2F transcription factors and advancement into S phase. Here we describe two simple techniques used to assess pRB phosphorylation and E2F transcription during G1/S progression.
G1/S期限制点是哺乳动物细胞分裂周期中的一个重要标志。G1/S期转换的关键调节因子是视网膜母细胞瘤基因产物（pRB）。它通过抑制E2F转录因子来阻止S期进展所需基因的转录。pRB的细胞周期蛋白依赖性激酶（Cdk）磷酸化增加会导致E2F转录因子释放并进入S期。在这里，我们描述了两种用于评估G1/S期进展过程中pRB磷酸化和E2F转录的简单技术。
