Journal
METHODS
Volume 86, Issue -, Pages 19-26Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymeth.2015.04.033
Keywords
RNA polymerase; Transcription initiation; Transcription regulation; Fluorescence spectroscopy; RNA polymerase promoter interactions
Funding
- National Institutes of Health [R01 GM59295]
- Molecular and Cell Biology Program grant from the Russian Academy of Sciences Presidium
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The molecular details of formation of transcription initiation complex upon the interaction of bacterial RNA polymerase (RNAP) with promoters are not completely understood. One way to address this problem is to understand how RNAP interacts with different parts of promoter DNA. A recently developed fluorometric RNAP molecular beacon assay allows one to monitor the RNAP interactions with various unlabeled DNA probes and quantitatively characterize partial RNAP-promoter interactions. This paper focuses on methodological aspects of application of this powerful assay to study the mechanism of transcription initiation complex formation by Escherichia coli RNA polymerase sigma(70) holoenzyme and its regulation by bacterial and phage encoded factors. (C) 2015 Elsevier Inc. All rights reserved.
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