Active establishment of centromeric CENP-A chromatin by RSF complex.

Active establishment of centromeric CENP-A chromatin by RSF complex.
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DOI:
10.1083/jcb.200903088
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发表时间:
2009-05-04
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Yoda K
Yoda K
中科院分区:
其他
文献类型:
--
作者:
Perpelescu M;Nozaki N;Obuse C;Yang H;Yoda K

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着丝粒是与子细胞同等DNA分离所需的染色体结构,包括含有着丝粒蛋白A (CENP-A)组蛋白的特殊核小体,它为着丝粒染色质组装提供了基础。着丝粒蛋白成分的发现正在取得进展,但有关其建立和维持的知识仍然有限。以前,我们使用抗cenp - a天然染色质免疫沉淀,分离了间期着丝粒复合体(ICEN)。在ICEN组分中,发现了重塑和间隔因子(RSF)复合物的亚基,即RSF -1和SNF2h蛋白。本文描述了RSF复合物与着丝粒结构和功能的关系,证明了其在HeLa细胞中对着丝粒核心染色质上CENP-A的维持需要。RSF复合物在g1中期与CENP-A染色质相互作用。rf -1缺失诱导着丝粒CENP-A丢失,纯化的RSF复合物在体外重组并间隔CENP-A核小体。根据这些数据,我们提出RSF复合体是一个积极支持CENP-A染色质组装的新因子。
Centromeres are chromosomal structures required for equal DNA segregation to daughter cells, comprising specialized nucleosomes containing centromere protein A (CENP-A) histone, which provide the basis for centromeric chromatin assembly. Discovery of centromere protein components is progressing, but knowledge related to their establishment and maintenance remains limited. Previously, using anti-CENP-A native chromatin immunoprecipitation, we isolated the interphase–centromere complex (ICEN). Among ICEN components, subunits of the remodeling and spacing factor (RSF) complex, Rsf-1 and SNF2h proteins, were found. This paper describes the relationship of the RSF complex to centromere structure and function, demonstrating its requirement for maintenance of CENP-A at the centromeric core chromatin in HeLa cells. The RSF complex interacted with CENP-A chromatin in mid-G1. Rsf-1 depletion induced loss of centromeric CENP-A, and purified RSF complex reconstituted and spaced CENP-A nucleosomes in vitro. From these data, we propose the RSF complex as a new factor actively supporting the assembly of CENP-A chromatin.
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