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首页> 外文期刊>Molecular and Cellular Biology >Differential Requirement of SAGA Components for Recruitment of TATA-Box-Binding Protein to Promoters In Vivo
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Differential Requirement of SAGA Components for Recruitment of TATA-Box-Binding Protein to Promoters In Vivo

机译：SAGA组件对TATA盒结合蛋白在体内向启动子的募集的差异要求

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The multisubunit Saccharomyces cerevisiae SAGA (Spt-Ada-Gcn5-acetyltransferase) complex is required to activate transcription of a subset of RNA polymerase II-dependent genes. However, the contribution of each SAGA component to transcription activation is relatively unknown. Here, using a formaldehyde-based in vivo cross-linking and chromatin immunoprecipitation assay, we have systematically analyzed the role of SAGA components in the recruitment of TATA-box binding protein (TBP) to SAGA-dependent promoters. We show that recruitment of TBP is diminished at a number of SAGA-dependent promoters in ada1Δ, spt7Δ, and spt20Δ null mutants, consistent with previous biochemical data suggesting that these components maintain the integrity of the SAGA complex. We also find that Spt3p is generally required for TBP binding to SAGA-dependent promoters, consistent with biochemical and genetic experiments, suggesting that Spt3p interacts with and recruits TBP to the core promoter. By contrast, Spt8p, which has been proposed to be required for the interaction between Spt3p and TBP, is required for TBP binding at only a subset of SAGA-dependent promoters. Ada2p and Ada3p are both required for TBP recruitment to Gcn5p-dependent promoters, supporting previous biochemical data that Ada2p and Ada3p are required for the histone acetyltransferase activity of Gcn5p. Finally, our results suggest that TBP-associated-factor components of SAGA are differentially required for TBP binding to SAGA-dependent promoters. In summary, we show that SAGA-dependent promoters require different combinations of SAGA components for TBP recruitment, revealing a complex combinatorial network for transcription activation in vivo.
机译：需要多亚基啤酒酵母SAGA（Spt-Ada-Gcn5-乙酰基转移酶）复合物来激活RNA聚合酶II依赖性基因的一个子集的转录。但是，每个SAGA组件对转录激活的贡献是相对未知的。在这里，我们使用基于甲醛的体内交联和染色质免疫沉淀测定法，系统地分析了SAGA组分在TATA盒结合蛋白（TBP）募集到SAGA依赖性启动子中的作用。我们显示，在 ada1 Δ， spt7 Δ和 spt20 Δnull突变体的许多SAGA依赖性启动子上，TBP的募集减少了，与以前的生化数据一致，表明这些成分保持了SAGA复合物的完整性。我们还发现，与生物化学和遗传实验一致，TBP结合SAGA依赖性启动子通常需要Spt3p，这表明Spt3p与TBP相互作用并将其招募到核心启动子上。相比之下，已经提议Spt3p和TBP之间的相互作用所必需的Spt8p仅在SAGA依赖性启动子的一个子集上需要TBP结合。 TBP募集至Gcn5p依赖性启动子均需要Ada2p和Ada3p，从而支持以前的生化数据，即Gcn5p的组蛋白乙酰转移酶活性需要Ada2p和Ada3p。最后，我们的结果表明，SABP的TBP相关因子组分是TBP与SAGA依赖的启动子结合的差异所需的。总之，我们表明依赖SAGA的启动子需要不同的SAGA组件组合来进行TBP募集，从而揭示了体内转录激活的复杂组合网络。

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