Generation of the dominant-negative mutant of hArpNβ:: A component of human SWI/SNF chromatin remodeling complex
Generation of the dominant-negative mutant of hArpNβ:: A component of human SWI/SNF chromatin remodeling complex
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DOI：
10.1006/excr.2001.5355
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发表时间：
2001-11-15
影响因子：
3.7
通讯作者：
Kwon, H
中科院分区：
文献类型：
--
作者：
Choi, EY;Park, JA;Kwon, H
hArpN beta 是一种位于细胞核内的肌动蛋白相关蛋白，是人类 SWI/SNF 染色质重塑复合物的一个亚基。 hArpN beta 已被提议调节 hSWI/SNF 复合物的组装和活性。潜在的 ArpN 同源物与 β-肌动蛋白的序列比较表明，除了独特的 N 端短插入片段 MS(G/A)-(V/L)YGG 之外，ArpN 还具有不同的子结构域 Ib 和 IIb。由于 hArpN beta 的拟议功能需要两个以上不同但同时的操作表面，因此我们检查了 hArpN beta 的一个操作表面的破坏是否会导致显性失活表型。当在 HeLa 或 293T 细胞中过表达时，亚结构域 Ib 或 IIb 杂合体（其中 hArpN β 的亚结构域 Ib 或 IIb 分别被 β-肌动蛋白的亚结构域替换）显示对细胞存活没有影响。另一方面，hArpN beta N端缺失突变体的过度表达可能通过细胞凋亡过程导致细胞死亡。这些结果表明 hArpN beta 的正常功能对于人类细胞的存活至关重要。此外，他们还表明，在 hArpN beta 与功能必需的伴侣蛋白结合后，N 端短序列对于染色质重塑活性或 hSWI/SNF 复合物的组装是必不可少的。 (C) 2001 年学术出版社。
hArpN beta, an actin-related protein located within the nucleus, is a subunit of the human SWI/SNF chromatin remodeling complex. hArpN beta has been proposed to regulate the assembly and activity of the hSWI/SNF complex. Sequence comparisons of the potential ArpN homologs with beta -actin showed that the ArpNs have the divergent subdomains Ib and IIb in addition to the unique N-terminal short insert, MS(G/A)-(V/L)YGG. Since the proposed function of hArpN beta requires more than two distinct but concurrently operating surfaces, we examined whether the disruption of one operating surface of hArpN beta results in dominant-negative phenotype. When overexpressed in HeLa or 293T cells, the subdomain Ib or IIb hybrids, in which the subdomain Ib or IIb of hArpN beta was replaced with that of beta -actin, respectively, showed no effect on cell survival. On the other hand, the overexpression of the N-terminal deletion mutant of hArpN beta resulted in cell death probably through apoptotic process. These results indicate that the proper function of hArpN beta is essential for cell survival in human cells. Furthermore, they suggests the possibility that the N-terminal short sequence is indispensable for the chromatin remodeling activity or the assembly of the hSWI/SNF complex after the binding of hArpN beta with functionally essential partner proteins. (C) 2001 Academic Press.