In vitro and in vivo methodologies for studying the Sigma 54-dependent transcription.

Martin Buck, Christoph Engl, Nicolas Joly, Goran Jovanovic, Edward Lawton, Christopher McDonald, Jorg Schumacher, Christopher J. Waite, Nan Zhang

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)

Abstract

Here we describe approaches and methods to assaying in vitro the major variant bacterial sigma factor, Sigma 54 (σ54), in a purified system. We include the complete transcription system, binding interactions between σ54 and its activators, as well as the self-assembly and the critical ATPase activity of the cognate activators which serve to remodel the closed promoter complexes. We also present in vivo methodologies that are used to study the impact of physiological processes, metabolic states, global signalling networks, and cellular architecture on the control of σ54-dependent gene expression.
Original languageEnglish
Title of host publicationBacterial Transcriptional Control: methods and protocols
EditorsIrina Artsimovitch, Thomas J Santangelo
Place of PublicationNew York
PublisherSpringer
Pages53-79
Volume1276
ISBN (Electronic)9781493923922
ISBN (Print)9781493923915
DOIs
Publication statusPublished - 28 Jan 2015

Publication series

NameMethods in Molecular Biology
PublisherSpringer New York
ISSN (Print)1064-3745

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  • Cite this

    Buck, M., Engl, C., Joly, N., Jovanovic, G., Lawton, E., McDonald, C., Schumacher, J., Waite, C. J., & Zhang, N. (2015). In vitro and in vivo methodologies for studying the Sigma 54-dependent transcription. In I. Artsimovitch, & T. J. Santangelo (Eds.), Bacterial Transcriptional Control: methods and protocols (Vol. 1276, pp. 53-79). (Methods in Molecular Biology). Springer. https://doi.org/10.1007/978-1-4939-2392-2_4