Promoter Independent Abortive Transcription Assays Unravel Functional Interactions Between TFIIB and RNA Polymerase
File(s)Methods in Molecular Biology_977_2013.pdf (586.88 KB)
Accepted version
Author(s)
Wiesler, SC
Werner, F
Weinzierl, RO
Type
Journal Article
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.
Date Issued
2013
Citation
Methods Mol Biol, 2013, 977, pp.217-227
ISSN
1940-6029
Start Page
217
End Page
227
Journal / Book Title
Methods Mol Biol
Volume
977
Copyright Statement
© Springer Science+Business Media, LLC 2013
Identifier
http://www.ncbi.nlm.nih.gov/pubmed/23436365
Coverage Spatial
United States