Hinojos Cruz A D, Sharp Z Dave, Mancini Michael A
Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston TX 77030, USA.
Trends Endocrinol Metab. 2005 Jan-Feb;16(1):12-8. doi: 10.1016/j.tem.2004.11.006.
The development of live cell and biochemical analysis methods has led to an increase in our understanding of the dynamic regulation of transcription. Live single cell studies using photobleaching techniques indicate that many proteins have a high nuclear mobility. Pioneering work using promoter array systems based on the lac operon or the mouse mammary tumor virus promoter enabled the study of chromatin structure, promoter occupancy and protein-chromatin interaction dynamics in relation to transcription. Chromatin immunoprecipitation (ChIP)-based assays allow an exhaustive analysis of the temporal recruitment of proteins to an endogenous promoter and provide evidence of cyclic protein-protein and protein-promoter interactions. Although reflecting different timescales, both ChIP and live cell studies indicate a highly dynamic control of transcription that until now has gone undetected and unappreciated.
活细胞和生化分析方法的发展使我们对转录的动态调控有了更深入的了解。使用光漂白技术的活单细胞研究表明,许多蛋白质具有较高的核迁移率。基于乳糖操纵子或小鼠乳腺肿瘤病毒启动子的启动子阵列系统的开创性工作,使得人们能够研究与转录相关的染色质结构、启动子占据情况以及蛋白质-染色质相互作用动力学。基于染色质免疫沉淀(ChIP)的分析方法能够详尽地分析蛋白质向内源启动子的时间募集情况,并提供蛋白质-蛋白质和蛋白质-启动子循环相互作用的证据。尽管ChIP和活细胞研究反映的时间尺度不同,但二者均表明转录存在高度动态的调控,而这一点至今尚未被发现和重视。