动粒组蛋白折叠复合体的生化与结构分析

Biochemical and Structural Analysis of Kinetochore Histone-Fold Complexes.

作者信息

Nishino Tatsuya, Fukagawa Tatsuo

机构信息

Department of Molecular Genetics, National Institute of Genetics and Graduate University for Advanced Studies (SOKENDAI), 1111 Yata, Mishima, Shizuoka, 411-8540, Japan.

Department of Biological Science and Technology, Graduate School of Industrial Science and Technology, Tokyo University of Science, Katsushika-ku, Tokyo, 125-8585, Japan.

出版信息

Methods Mol Biol. 2016;1413:135-46. doi: 10.1007/978-1-4939-3542-0_9.

DOI:10.1007/978-1-4939-3542-0_9

PMID:27193847

Abstract

The kinetochore connects chromosomes to microtubules during mitosis and therefore plays an essential role in faithful chromosome segregation. It is built at the centromeric region of the chromosome and is comprised of many protein complexes. CENP-S, -T, -W, and -X are kinetochore components with histone-folds. These proteins play important roles in establishment of kinetochore chromatin. Similar to canonical histones, these kinetochore histone-fold proteins form heteromeric complexes (CENP-S/CENP-X complex and CENP-T/CENP-W complex) and bind DNA in sequence independent manner. In addition, they form a CENP-T-W-S-X heterotetrameric complex and bind DNA in a manner that is different from both CENP-S-X and CENP-T-W. To understand how kinetochores form and function it is necessary to characterize the components in detail. Here, we describe our approaches in purification and characterization of the kinetochore histone-fold complexes.

摘要

在有丝分裂过程中，动粒将染色体与微管相连，因此在准确的染色体分离中起着至关重要的作用。它在染色体的着丝粒区域构建，由许多蛋白质复合物组成。CENP-S、-T、-W和-X是具有组蛋白折叠结构的动粒组分。这些蛋白质在动粒染色质的建立中发挥重要作用。与经典组蛋白相似，这些动粒组蛋白折叠蛋白形成异源复合物（CENP-S/CENP-X复合物和CENP-T/CENP-W复合物），并以序列非依赖性方式结合DNA。此外，它们形成CENP-T-W-S-X异源四聚体复合物，并以不同于CENP-S-X和CENP-T-W的方式结合DNA。为了了解动粒如何形成和发挥功能，有必要详细表征其组分。在这里，我们描述了我们纯化和表征动粒组蛋白折叠复合物的方法。