Armache Karim-Jean, Mitterweger Simone, Meinhart Anton, Cramer Patrick
Gene Center, University of Munich (LMU), Department of Chemistry and Biochemistry, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.
J Biol Chem. 2005 Feb 25;280(8):7131-4. doi: 10.1074/jbc.M413038200. Epub 2004 Dec 9.
We determined the x-ray structure of the RNA polymerase (Pol) II subcomplex Rpb4/7 at 2.3 A resolution, combined it with a previous structure of the 10-subunit polymerase core, and refined an atomic model of the complete 12-subunit Pol II at 3.8-A resolution. Comparison of the complete Pol II structure with structures of the Pol II core and free Rpb4/7 shows that the core-Rpb4/7 interaction goes along with formation of an alpha-helix in the linker region of the largest Pol II subunit and with folding of the conserved Rpb7 tip loop. Details of the core-Rpb4/7 interface explain facilitated Rpb4/7 dissociation in a temperature-sensitive Pol II mutant and specific assembly of Pol I with its Rpb4/7 counterpart, A43/14. The refined atomic model of Pol II serves as the new reference structure for analysis of the transcription mechanism and enables structure solution of complexes of the complete enzyme with additional factors and nucleic acids by molecular replacement.
我们确定了RNA聚合酶(Pol)II亚复合物Rpb4/7在2.3埃分辨率下的X射线结构,将其与先前的10亚基聚合酶核心结构相结合,并在3.8埃分辨率下优化了完整的12亚基Pol II的原子模型。将完整的Pol II结构与Pol II核心结构及游离的Rpb4/7结构进行比较,结果表明核心与Rpb4/7的相互作用伴随着最大的Pol II亚基连接区α螺旋的形成以及保守的Rpb7末端环的折叠。核心与Rpb4/7界面的细节解释了温度敏感型Pol II突变体中Rpb4/7解离的促进作用以及Pol I与其对应物A43/14的特异性组装。优化后的Pol II原子模型为转录机制分析提供了新的参考结构,并能够通过分子置换解析完整酶与其他因子及核酸复合物的结构。