Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
J Biol Chem. 2010 Oct 29;285(44):33671-9. doi: 10.1074/jbc.M110.140764. Epub 2010 Aug 23.
The Paf1 complex (Paf1C) affects RNA polymerase II transcription by coordinating co-transcriptional chromatin modifications and helping recruit mRNA 3' end processing factors. Paf1C cross-links to transcribed genes, but not downstream of the cleavage and polyadenylation site, suggesting that it may interact with the nascent mRNA. Paf1C purified from Saccharomyces cerevisiae binds RNA in vitro, as do the purified Leo1 and Rtf1 subunits of the complex. In vivo cross-linking and immunoprecipitation of RNA associated with Paf1C (RNA-IP) show that Leo1, but not Rtf1, is necessary for the complex to bind RNA. Cells lacking Leo1 have reduced Paf1C recruitment as well as decreased levels of histone H3 and trimethylated H3 Lys(4) within transcribed chromatin. Together, these results suggest that association of Paf1C with RNA stabilizes its localization at actively transcribed regions where it influences chromatin structure.
Paf1 复合物 (Paf1C) 通过协调共转录染色质修饰并帮助招募 mRNA 3'端加工因子来影响 RNA 聚合酶 II 转录。Paf1C 与转录基因交联,但不在切割和多聚腺苷酸化位点的下游,表明它可能与新生 mRNA 相互作用。从酿酒酵母中纯化的 Paf1C 在体外结合 RNA,复合物的纯化 Leo1 和 Rtf1 亚基也是如此。与 Paf1C(RNA-IP)相关的体内交联和免疫沉淀显示,Leo1 而不是 Rtf1 对于复合物结合 RNA 是必需的。缺乏 Leo1 的细胞中,Paf1C 的募集减少,以及转录染色质中组蛋白 H3 和三甲基化 H3 Lys(4)的水平降低。这些结果表明,Paf1C 与 RNA 的结合稳定了其在活跃转录区域的定位,在这些区域,它影响染色质结构。