title: Promoter independent abortive transcription assays unravel functional interactions between TFIIB and RNA polymerase.
abstract: TFIIB-like general transcription factors are required for transcription initiation by all eukaryotic and archaeal RNA polymerases (RNAPs). TFIIB facilitates both recruitment and post-recruitment steps of initiation; in particular, TFIIB stimulates abortive initiation. X-ray crystallography of TFIIB-RNAP II complexes shows that the TFIIB linker region penetrates the RNAP active center, yet the impact of this arrangement on RNAP activity and underlying mechanisms remains elusive. Promoter-independent abortive initiation assays exploit the intrinsic ability of RNAP enzymes to initiate transcription from nicked DNA templates and record the formation of the first phosphodiester bonds. These assays can be used to measure the effect of transcription factors such as TFIIB and RNAP mutations on abortive transcription.
authors: Department of Life Sciences, Imperial College London, London, UK.
authors: Simone C
authors: SC
authors: Wiesler
authors: Finn
authors: F
authors: Werner
authors: Robert O J
authors: RO
authors: Weinzierl
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry*; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry; D017930:Genes, Reporter; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D017015:Methanococcales / Q000201:enzymology / Q000235:genetics; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017186:Titrimetry; D035581:Transcription Factor TFIIB / Q000737:chemistry*; D061785:Transcription Initiation, Genetic; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000737:chemistry*
chemical_list: D019843:Archaeal Proteins; D029968:Escherichia coli Proteins; D035581:Transcription Factor TFIIB; D014764:Viral Proteins; D008156:Luciferases; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases