Coulombe Benoit, Langelier Marie-France
Gene Transcription Laboratory, Institute de recherche cliniques de Montréal, QC, Canada.
Biochem Cell Biol. 2005 Aug;83(4):497-504. doi: 10.1139/o05-061.
High resolution X-ray crystal structures of multisubunit RNA polymerases (RNAP) have contributed to our understanding of transcriptional mechanisms. They also provided a powerful guide for the design of experiments aimed at further characterizing the molecular stages of the transcription reaction. Our laboratory used tandem-affinity peptide purification in native conditions to isolate human RNAP II variants that had site-specific mutations in structural elements located strategically within the enzyme's catalytic center. Both in vitro and in vivo analyses of these mutants revealed novel features of the catalytic mechanisms involving this enzyme.
多亚基RNA聚合酶（RNAP）的高分辨率X射线晶体结构有助于我们理解转录机制。它们还为旨在进一步表征转录反应分子阶段的实验设计提供了有力指导。我们实验室在天然条件下使用串联亲和肽纯化法，分离出在酶催化中心内战略性定位的结构元件中具有位点特异性突变的人RNAP II变体。对这些突变体的体外和体内分析揭示了涉及该酶的催化机制的新特征。
