不同的负延伸因子构象调节 RNA 聚合酶 II 启动子近端暂停。
Distinct negative elongation factor conformations regulate RNA polymerase II promoter-proximal pausing.
机构信息
Department of Biology, Massachusetts Institute of Technology, Building 68, 31 Ames St., Cambridge, MA 02139, USA.
Department of Biology, Massachusetts Institute of Technology, Building 68, 31 Ames St., Cambridge, MA 02139, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA.
出版信息
Mol Cell. 2024 Apr 4;84(7):1243-1256.e5. doi: 10.1016/j.molcel.2024.01.023. Epub 2024 Feb 23.
Metazoan gene expression regulation involves pausing of RNA polymerase (Pol II) in the promoter-proximal region of genes and is stabilized by DSIF and NELF. Upon depletion of elongation factors, NELF appears to accompany elongating Pol II past pause sites; however, prior work indicates that NELF prevents Pol II elongation. Here, we report cryoelectron microscopy structures of Pol II-DSIF-NELF complexes with NELF in two distinct conformations corresponding to paused and poised states. The paused NELF state supports Pol II stalling, whereas the poised NELF state enables transcription elongation as it does not support a tilted RNA-DNA hybrid. Further, the poised NELF state can accommodate TFIIS binding to Pol II, allowing for Pol II reactivation at paused or backtracking sites. Finally, we observe that the NELF-A tentacle interacts with the RPB2 protrusion and is necessary for pausing. Our results define how NELF can support pausing, reactivation, and elongation by Pol II.
后生动物基因表达调控涉及 RNA 聚合酶(Pol II)在基因启动子近端区域的暂停,并由 DSIF 和 NELF 稳定。在伸长因子耗尽的情况下,NELF 似乎伴随着延伸的 Pol II 越过暂停位点;然而,之前的工作表明 NELF 阻止 Pol II 的延伸。在这里,我们报告了 Pol II-DSIF-NELF 复合物的冷冻电子显微镜结构,其中 NELF 处于两种不同的构象,分别对应于暂停和准备状态。暂停的 NELF 状态支持 Pol II 的停顿,而准备的 NELF 状态能够支持转录延伸,因为它不支持倾斜的 RNA-DNA 杂交。此外,准备的 NELF 状态可以容纳 TFIIS 与 Pol II 的结合,从而允许 Pol II 在暂停或回溯位点重新激活。最后,我们观察到 NELF-A 触须与 RPB2 突出部相互作用,这对于暂停是必要的。我们的结果定义了 NELF 如何支持 Pol II 的暂停、重新激活和延伸。