权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Constitution and Function of human SWI/SNF Complex : Implication in supression of carcionogenesis

人类 SWI/SNF 复合物的构成和功能：抑制癌发生的意义

基本信息

批准号：
14580657
负责人：
FURUKAWA Takako
金额：
$ 1.22万
依托单位：
Faculty of Medical Sciences, University of Fukui
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580657/
关键词：
chromatin remodeling SWI SNF Osa chromatin remodeling complex 転写調節

项目摘要

We isolated two related but distinct cDNA clones, hOsa-1 and hOsa-2, the 1argest component of human SWI/SNF complex. They had ARID (AT-rich interaction domain) and two high homology region to drosophila Osa. Co-immunoprecipitation study revealed that these C-terminal homology regions were important for the interaction with BRG and BRM, ATP-dependent enzyme componens of SWI/SNF. In cultured cells hOsa stimulated transcription by steroid hormone receptors. ARID lacking mutant did not stimulate the transcription, suggesting the importance of this domain in SWI/SNF function. We established HeLa cell lines which express either wild type or a mutant missing the ARID in Flag-tagged form under the control of tetracycline administration. Putative human SWI/SNF components, Osa, BRG, BAF155, were found in the high molecular weight fractions when the nuclear extracts were separated by glycerol gradient sedimentation. Distribution of BAF170 was different between the extracts from wild type and the mutant expressing cells. There might be some interaction between BAF170 and ARID of Osa-2. Silver staining of the complexes with wild type and the mutant Osa-2 immunopurified from the high molecular weight sedimentation fractions showed quite similar pattern, suggesting the presence of ARID has little effect on the complex formation.

我们分离了两个相关但不同的cDNA克隆，hOsa-1和hOsa-2，它们是人SWI/SNF复合物的最大组分。它们具有ARID（AT-rich interaction domain）和两个与果蝇Osa高度同源的区域。免疫共沉淀研究表明，这些C-末端同源区域对于SWI/SNF的ATP依赖性酶组分BRG和BRM的相互作用是重要的。在培养的细胞中，hOsa通过类固醇激素受体刺激转录。ARID缺失突变体不能刺激转录，提示该结构域在SWI/SNF功能中的重要性。我们建立了HeLa细胞系，表达野生型或突变体缺失的ARID在标记的形式下四环素管理的控制下。当用甘油梯度沉降法分离核提取物时，在高分子量组分中发现了推定的人SWI/SNF组分Osa、BRG、BAF 155。BAF 170在野生型和突变型表达细胞提取物中的分布不同。BAF 170与Osa-2的ARID可能存在一定的相互作用。从高分子量沉降组分中免疫纯化的野生型和突变型Osa-2的复合物的银染色显示出非常相似的模式，表明ARID的存在对复合物的形成几乎没有影响。