Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, 108 Althouse Laboratory, Pennsylvania State University, University Park, PA 16802-1014, USA.
Biochem Soc Trans. 2012 Apr;40(2):351-6. doi: 10.1042/BST20110734.
The nucleosome repeating unit of chromatin is the target of chromatin enzymes and factors that regulate gene activity in a eukaryotic cell. How the nucleosome is recognized by chromatin enzymes and factors is poorly understood, even though such interaction is fundamental to gene regulation and chromatin biology. My laboratory recently determined the structural basis for how the RCC1 (regulator of chromosome condensation 1) chromatin factor binds to the nucleosome, including the first atomic crystal structure of a chromatin protein complexed with the nucleosome core particle. I describe here how we developed and investigated structural models for RCC1 binding to the nucleosome using biochemical methods and how we crystallized the 300 kDa complex of RCC1 with the nucleosome core particle. This article highlights the contributions made by key laboratory members and explains our thinking and rationale during the discovery process.
染色质的核小体重复单元是调节真核细胞基因活性的染色质酶和因子的作用靶点。尽管这种相互作用是基因调控和染色质生物学的基础,但核小体如何被染色质酶和因子识别仍知之甚少。我的实验室最近确定了 RCC1(染色体凝聚调节因子 1)染色质因子与核小体结合的结构基础,包括第一个与核小体核心颗粒结合的染色质蛋白复合物的原子晶体结构。我在这里描述了我们如何使用生化方法开发和研究 RCC1 与核小体结合的结构模型,以及我们如何结晶出 RCC1 与核小体核心颗粒的 300 kDa 复合物。本文重点介绍了实验室关键成员的贡献,并解释了我们在发现过程中的思考和基本原理。