Ferreira Monica E, Prochasson Philippe, Berndt Kurt D, Workman Jerry L, Wright Anthony P H
School of Life Sciences, Södertörns Högskola, Huddinge, Sweden.
FEBS J. 2009 May;276(9):2557-65. doi: 10.1111/j.1742-4658.2009.06979.x. Epub 2009 Mar 18.
Interaction between acidic activation domains and the activator-binding domains of Swi1 and Snf5 of the yeast SWI/SNF chromatin remodeling complex has previously been characterized in vitro. Although deletion of both activator-binding domains leads to phenotypes that differ from the wild-type, their relative importance for SWI/SNF recruitment to target genes has not been investigated. In the present study, we used chromatin immunoprecipitation assays to investigate the individual and collective importance of the activator-binding domains for SWI/SNF recruitment to genes within the GAL regulon in vivo. We also investigated the consequences of defective SWI/SNF recruitment for target gene activation. We demonstrate that deletion of both activator-binding domains essentially abolishes galactose-induced SWI/SNF recruitment and causes a reduction in transcriptional activation similar in magnitude to that associated with a complete loss of SWI/SNF activity. The activator-binding domains in Swi1 and Snf5 make approximately equal contributions to the recruitment of SWI/SNF to each of the genes studied. The requirement for SWI/SNF recruitment correlates with GAL genes that are highly and rapidly induced by galactose.
酵母SWI/SNF染色质重塑复合物的酸性激活结构域与Swi1和Snf5的激活剂结合结构域之间的相互作用此前已在体外进行了表征。虽然两个激活剂结合结构域的缺失会导致与野生型不同的表型,但它们对SWI/SNF募集到靶基因的相对重要性尚未得到研究。在本研究中,我们使用染色质免疫沉淀测定法来研究激活剂结合结构域对体内SWI/SNF募集到GAL调控子内基因的个体和集体重要性。我们还研究了SWI/SNF募集缺陷对靶基因激活的影响。我们证明,两个激活剂结合结构域的缺失基本上消除了半乳糖诱导的SWI/SNF募集,并导致转录激活的降低,其幅度与与SWI/SNF活性完全丧失相关的降低相似。Swi1和Snf5中的激活剂结合结构域对SWI/SNF募集到每个研究基因的贡献大致相等。对SWI/SNF募集的需求与由半乳糖高度和快速诱导的GAL基因相关。