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对RSC58进行基因分析,RSC58编码酵母染色质重塑复合物的一个组分,并与转录因子Swi6相互作用。

Genetic analysis of RSC58, which encodes a component of a yeast chromatin remodeling complex, and interacts with the transcription factor Swi6.

作者信息

Taneda T, Kikuchi A

机构信息

Division of Molecular Mycology and Medicine, Center for Neural Disease and Cancer, Graduate School of Medicine, Nagoya University, Tsurumai-cho 65, Showa-ku, 466-8550 Nagoya, Japan.

出版信息

Mol Genet Genomics. 2004 May;271(4):479-89. doi: 10.1007/s00438-004-0999-3. Epub 2004 Mar 19.

Abstract

Before transcription can begin, the chromatin structure must be rearranged at the nucleosome level. In yeast the nucleosome remodeling complex RSC is involved in this process and it is essential for growth. Recent analysis of the RSC by mass spectrometry has suggested that the product of YLR033w, an essential gene of unknown function, is a novel component of the complex, and the gene has been renamed RSC58. Rsc58 is predicted to be 502 amino acids long. We have isolated five temperature-sensitive mutations in RSC58 and studied the cellular function of the gene. Our major findings are the following. (1) Two of the alleles have a frameshift mutation near the 3' end of the gene, in codons 482 and 485, respectively. The first mutation is associated with the more severe phenotype. This is compatible with the finding that removal of the C-terminal 25 residues of Rsc58 is lethal to cells. These results suggest that C-terminal region is essential for Rsc58 function. (2) RSC4, which codes for another member of the RSC, was found to be a multicopy suppressor of the phenotype of one of the temperature-sensitive mutants. (3) Two-hybrid analysis identified Swi6, a transcription factor, as a candidate interactor with Rsc58. An interaction between Rsc58 and Swi6 is also suggested by the fact that rsc58-ts Deltaswi6 double mutants show a more severe growth defect than either mutation alone. These results suggest the possibility that Rsc58 mediates between nucleosome remodeling and the initiation of transcription.

摘要

在转录开始之前,染色质结构必须在核小体水平上进行重排。在酵母中,核小体重塑复合物RSC参与了这一过程,并且对生长至关重要。最近通过质谱对RSC进行的分析表明,YLR033w(一个功能未知的必需基因)的产物是该复合物的一个新组分,该基因已被重新命名为RSC58。预测Rsc58由502个氨基酸组成。我们分离出了RSC58中的五个温度敏感突变,并研究了该基因的细胞功能。我们的主要发现如下:(1)其中两个等位基因在基因的3'端附近分别在密码子482和485处发生了移码突变。第一个突变与更严重的表型相关。这与以下发现一致,即去除Rsc58的C末端25个残基对细胞是致死的。这些结果表明C末端区域对Rsc58的功能至关重要。(2)编码RSC另一个成员的RSC4被发现是其中一个温度敏感突变体表型的多拷贝抑制子。(3)双杂交分析确定转录因子Swi6是与Rsc58相互作用的候选蛋白。rsc58-ts Deltaswi6双突变体表现出比单独任何一个突变更严重的生长缺陷,这一事实也表明Rsc58与Swi6之间存在相互作用。这些结果表明Rsc58可能在核小体重塑和转录起始之间起介导作用。

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