Simic Rajna, Lindstrom Derek L, Tran Hien G, Roinick Kelli L, Costa Patrick J, Johnson Alexander D, Hartzog Grant A, Arndt Karen M
Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260, USA.
EMBO J. 2003 Apr 15;22(8):1846-56. doi: 10.1093/emboj/cdg179.
Transcription in eukaryotes is influenced by the chromatin state of the template, and chromatin remodeling factors have well-documented roles in regulating transcription initiation by RNA polymerase (pol) II. Chromatin also influences transcription elongation; however, little is known about the role of chromatin remodeling factors in this process. Here, we present evidence that the Saccharomyces cerevisiae chromatin remodeling factor Chd1 functions during transcription elongation. First, we identified Chd1 in a two-hybrid screen for proteins that interact with Rtf1, a member of the Paf1 complex that associates with RNA pol II and regulates transcription elongation. Secondly, we show through co-immunoprecipitation studies that Chd1 also interacts with components of two essential elongation factors, Spt4-Spt5 and Spt16-Pob3. Thirdly, we demonstrate that deletion of CHD1 suppresses a cold-sensitive spt5 mutation that is also suppressed by defects in the Paf1 complex and RNA pol II. Finally, we demonstrate that Chd1, Rtf1 and Spt5 associate with actively transcribed regions of chromatin. Collectively, these findings suggest an important role for Chd1 and chromatin remodeling in the control of transcription elongation.
真核生物中的转录受模板染色质状态的影响,染色质重塑因子在调节RNA聚合酶(pol)II起始转录方面具有充分记录的作用。染色质也影响转录延伸;然而,关于染色质重塑因子在此过程中的作用知之甚少。在这里,我们提供证据表明酿酒酵母染色质重塑因子Chd1在转录延伸过程中发挥作用。首先,我们在一个双杂交筛选中鉴定出Chd1,该筛选针对与Rtf1相互作用的蛋白质,Rtf1是Paf1复合物的成员,与RNA pol II相关联并调节转录延伸。其次,我们通过免疫共沉淀研究表明Chd1也与两个必需延伸因子Spt4-Spt5和Spt16-Pob3的组分相互作用。第三,我们证明缺失CHD1可抑制冷敏感的spt5突变,Paf1复合物和RNA pol II中的缺陷也可抑制该突变。最后,我们证明Chd1、Rtf1和Spt5与染色质的活跃转录区域相关联。总的来说,这些发现表明Chd1和染色质重塑在转录延伸控制中具有重要作用。