Wang Xiaomei, Nagl Norman G, Wilsker Deborah, Van Scoy Michael, Pacchione Stephen, Yaciuk Peter, Dallas Peter B, Moran Elizabeth
Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, Philadelphia, PA 19140, USA.
Biochem J. 2004 Oct 15;383(Pt 2):319-25. doi: 10.1042/BJ20040524.
p270 (ARID1A) is a member of the ARID family of DNA-binding proteins and a subunit of human SWI/SNF-related complexes, which use the energy generated by an integral ATPase subunit to remodel chromatin. ARID1B is an independent gene product with an open reading frame that is more than 60% identical with p270. We have generated monoclonal antibodies specific for either p270 or ARID1B to facilitate the investigation of ARID1B and its potential interaction with human SWI/SNF complexes in vivo. Immunocomplex analysis provides direct evidence that endogenous ARID1B is associated with SWI/SNF-related complexes and indicates that p270 and ARID1B, similar to the ATPase subunits BRG1 and hBRM, are alternative, mutually exclusive subunits of the complexes. The ARID-containing subunits are not specific to the ATPases. Each associates with both BRG1 and hBRM, thus increasing the number of distinct subunit combinations known to be present in cells. Analysis of the panels of cell lines indicates that ARID1B, similar to p270, has a broad tissue distribution. The ratio of p270/ARID1B in typical cells is approx. 3.5:1, and BRG1 is distributed proportionally between the two ARID subunits. Analysis of DNA-binding behaviour indicates that ARID1B binds DNA in a non-sequence-specific manner similar to p270.
p270(ARID1A)是DNA结合蛋白ARID家族的成员,也是人类SWI/SNF相关复合物的一个亚基,该复合物利用一个完整的ATP酶亚基产生的能量来重塑染色质。ARID1B是一种独立的基因产物,其开放阅读框与p270的同源性超过60%。我们已经制备了针对p270或ARID1B的单克隆抗体,以促进对ARID1B及其在体内与人类SWI/SNF复合物潜在相互作用的研究。免疫复合物分析提供了直接证据,表明内源性ARID1B与SWI/SNF相关复合物有关联,并表明p270和ARID1B与ATP酶亚基BRG1和hBRM类似,是该复合物的可替代、相互排斥的亚基。含ARID的亚基并非ATP酶所特有。每个亚基都与BRG1和hBRM结合,从而增加了已知存在于细胞中的不同亚基组合的数量。对一系列细胞系的分析表明,ARID1B与p270类似,具有广泛的组织分布。典型细胞中p270/ARID1B的比例约为3.5:1,并且BRG1在两个ARID亚基之间按比例分布。对DNA结合行为的分析表明,ARID1B以类似于p270的非序列特异性方式结合DNA。