Nagaich Akhilesh K, Walker Dawn A, Wolford Ron, Hager Gordon L
Laboratory of Receptor Biology and Gene Expression, Building 41, B602, 41 Library Drive, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Mol Cell. 2004 Apr 23;14(2):163-74. doi: 10.1016/s1097-2765(04)00178-9.
An ultrafast UV laser crosslinking assay has provided novel insights into the progression of the SWI/SNF-mediated chromatin-remodeling reaction and transcription factor binding in real time. We demonstrate site-specific crosslinking between the glucocorticoid receptor (GR), the hSWI/SNF chromatin-remodeling complex, and the mouse mammary tumor virus (MMTV) promoter assembled in an array of correctly positioned nucleosomes. GR first demonstrates rapid binding to the promoter and then is actively displaced from the template during the remodeling reaction. This displacement reaction requires the hSWI/SNF complex and ATP, is specific to the nucleoprotein template, and is accompanied by a core histone rearrangement. The hSWI/SNF complex associates with random positions on the chromatin template in the absence of GR but is recruited specifically to the B/C region when GR is included. These results indicate that enhancement of hSWI/SNF-mediated factor accessibility, a hallmark of chromatin remodeling, is in some cases transient, reversible, and periodic.
一种超快紫外激光交联测定法为实时了解SWI/SNF介导的染色质重塑反应和转录因子结合的进程提供了新的见解。我们展示了糖皮质激素受体(GR)、hSWI/SNF染色质重塑复合物与组装在一系列正确定位核小体中的小鼠乳腺肿瘤病毒(MMTV)启动子之间的位点特异性交联。GR首先快速结合到启动子上,然后在重塑反应过程中从模板上被主动置换。这种置换反应需要hSWI/SNF复合物和ATP,对核蛋白模板具有特异性,并伴随着核心组蛋白重排。在没有GR的情况下,hSWI/SNF复合物与染色质模板上的随机位置结合,但当加入GR时,它会特异性地募集到B/C区域。这些结果表明,hSWI/SNF介导的因子可及性增强是染色质重塑的一个标志,在某些情况下是短暂的、可逆的和周期性的。