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首页> 外文期刊>Molecular and Cellular Biology >Drosophila Paf1 Modulates Chromatin Structure at Actively Transcribed Genes
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Drosophila Paf1 Modulates Chromatin Structure at Actively Transcribed Genes

机译：果蝇Paf1调节活跃转录基因的染色质结构。

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The Paf1 complex in yeast has been reported to influence a multitude of steps in gene expression through interactions with RNA polymerase II (Pol II) and chromatin-modifying complexes; however, it is unclear which of these many activities are primary functions of Paf1 and are conserved in metazoans. We have identified and characterized the Drosophila homologs of three subunits of the yeast Paf1 complex and found striking differences between the yeast and Drosophila Paf1 complexes. We demonstrate that although Drosophila Paf1, Rtf1, and Cdc73 colocalize broadly with actively transcribing, phosphorylated Pol II, and all are recruited to activated heat shock genes with similar kinetics; Rtf1 does not appear to be a stable part of the Drosophila Paf1 complex. RNA interference (RNAi)-mediated depletion of Paf1 or Rtf1 leads to defects in induction of Hsp70 RNA, but tandem RNAi-chromatin immunoprecipitation assays show that loss of neither Paf1 nor Rtf1 alters the density or distribution of phosphorylated Pol II on the active Hsp70 gene. However, depletion of Paf1 reduces trimethylation of histone H3 at lysine 4 in the Hsp70 promoter region and significantly decreases the recruitment of chromatin-associated factors Spt6 and FACT, suggesting that Paf1 may manifest its effects on transcription through modulating chromatin structure.
机译：据报道，酵母中的Paf1复合物通过与RNA聚合酶II（Pol II）和染色质修饰复合物的相互作用影响基因表达的多个步骤。但是，尚不清楚这些活动中的哪些是Paf1的主要功能，并且在后生动物中是否保守。我们已经鉴定并表征了酵母Paf1复合体的三个亚基的果蝇同源物，并发现了酵母与果蝇 Paf1复合体之间的显着差异。我们证明，尽管果蝇Paf1，Rtf1和Cdc73与活跃转录的磷酸化Pol II广泛共定位，并且全部被募集到具有相似动力学的活化热激基因。 Rtf1似乎不是果蝇Paf1复合体的稳定部分。 RNA干扰（RNAi）介导的Paf1或Rtf1耗竭会导致 Hsp70 RNA的诱导缺陷，但是串联RNAi-染色质免疫沉淀测定表明Paf1和Rtf1的丢失均不会改变磷酸化的密度或分布主动 Hsp70 基因上的Pol II。然而，Paf1的消耗减少了 Hsp70 启动子区域中赖氨酸4处组蛋白H3的三甲基化，并显着降低了染色质相关因子Spt6和FACT的募集，这表明Paf1可能通过调节来显示其对转录的影响染色质结构。

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