Functional dissection of the catalytic mechanism of mammalian RNA polymerase II.
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.
Coulombe
Benoit
B
Gene Transcription Laboratory, Institute de recherche cliniques de Montréal, QC, Canada. benoit.coulombe@ircm.qc.ca
Langelier
Marie-France
MF
D000818:Animals; D002384:Catalysis; D006801:Humans; D009154:Mutation; D010445:Peptide Elongation Factors / Q000502:physiology; D012319:RNA Polymerase II / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D019473:Transcription Factors, TFII / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014158:Transcription, Genetic
D010445:Peptide Elongation Factors; D019473:Transcription Factors, TFII; D012319:RNA Polymerase II