Coisy Marjorie, Roure Virginie, Ribot Magali, Philips Alexandre, Muchardt Christian, Blanchard Jean-Marie, Dantonel Jean-Christophe
Institut de Génétique Moléculaire, CNRS, UMR 5535, 1919 Route de Mende, 34293 Montpellier cedex 5, France.
Mol Cell. 2004 Jul 2;15(1):43-56. doi: 10.1016/j.molcel.2004.06.022.
Cell cycle-dependent expression of cyclin A is controlled by transcriptional repression in early phase of the cell cycle. In this study, we directly examine the chromatin structure of the mouse cyclin A promoter through in vivo micrococcal nuclease footprinting. We describe here that cyclin A repression is associated with two positioned nucleosomes and that histones progressively lose DNA contact synchronously with gene activation. This particular nucleosomal organization is disrupted by mutations of the cyclin A bipartite repressor sequence. Moreover, the same sequence recruits the chromatin remodeling factor Brahma/SNF2alpha (Brm) onto the cyclin A promoter. Accordingly, cyclin A proximal promoter is not wrapped around nucleosomes and not repressed in quiescent cells lacking Brm. These results provide molecular explanations for the transcriptional repression state of cyclin A, as well as insights into the action of Brm chromatin remodeling factor as cell cycle regulator.
细胞周期蛋白A的细胞周期依赖性表达在细胞周期早期受转录抑制调控。在本研究中,我们通过体内微球菌核酸酶足迹法直接检测小鼠细胞周期蛋白A启动子的染色质结构。我们在此描述,细胞周期蛋白A的抑制与两个定位核小体相关,并且组蛋白随着基因激活同步逐渐失去与DNA的接触。细胞周期蛋白A二分抑制序列的突变会破坏这种特定的核小体组织。此外,相同序列将染色质重塑因子Brahma/SNF2alpha(Brm)募集到细胞周期蛋白A启动子上。因此,在缺乏Brm的静止细胞中,细胞周期蛋白A近端启动子不缠绕在核小体上且不被抑制。这些结果为细胞周期蛋白A的转录抑制状态提供了分子解释,也为Brm染色质重塑因子作为细胞周期调节因子的作用提供了见解。