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人类着丝粒蛋白B诱导核小体在α-卫星序列上的翻译定位。

Human centromere protein B induces translational positioning of nucleosomes on alpha-satellite sequences.

作者信息

Tanaka Yoshinori, Tachiwana Hiroaki, Yoda Kinya, Masumoto Hiroshi, Okazaki Tsuneko, Kurumizaka Hitoshi, Yokoyama Shigeyuki

机构信息

Protein Research Group, RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045, Japan.

出版信息

J Biol Chem. 2005 Dec 16;280(50):41609-18. doi: 10.1074/jbc.M509666200. Epub 2005 Sep 23.

Abstract

The human centromere proteins A (CENP-A) and B (CENP-B) are the fundamental centromere components of chromosomes. CENP-A is the centromere-specific histone H3 variant, and CENP-B specifically binds a 17-base pair sequence (the CENP-B box), which appears within every other alpha-satellite DNA repeat. In the present study, we demonstrated centromere-specific nucleosome formation in vitro with recombinant proteins, including histones H2A, H2B, H4, CENP-A, and the DNA-binding domain of CENP-B. The CENP-A nucleosome wraps 147 base pairs of the alpha-satellite sequence within its nucleosome core particle, like the canonical H3 nucleosome. Surprisingly, CENP-B binds to nucleosomal DNA when the CENP-B box is wrapped within the nucleosome core particle and induces translational positioning of the nucleosome without affecting its rotational setting. This CENP-B-induced translational positioning only occurs when the CENP-B box sequence is settled in the proper rotational setting with respect to the histone octamer surface. Therefore, CENP-B may be a determinant for translational positioning of the centromere-specific nucleosomes through its binding to the nucleosomal CENP-B box.

摘要

人类着丝粒蛋白A(CENP-A)和B(CENP-B)是染色体的基本着丝粒成分。CENP-A是着丝粒特异性组蛋白H3变体,而CENP-B特异性结合一个17碱基对序列(CENP-B盒),该序列出现在每隔一个的α卫星DNA重复序列中。在本研究中,我们用重组蛋白在体外证明了着丝粒特异性核小体的形成,这些重组蛋白包括组蛋白H2A、H2B、H4、CENP-A以及CENP-B的DNA结合结构域。与典型的H3核小体一样,CENP-A核小体在其核小体核心颗粒内包裹147个碱基对的α卫星序列。令人惊讶的是,当CENP-B盒包裹在核小体核心颗粒内时,CENP-B会与核小体DNA结合,并诱导核小体的平移定位,而不影响其旋转设置。这种CENP-B诱导的平移定位仅在CENP-B盒序列相对于组蛋白八聚体表面处于适当的旋转设置时才会发生。因此,CENP-B可能通过与核小体CENP-B盒结合而成为着丝粒特异性核小体平移定位的决定因素。

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