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

Martin Buck; Christoph Engl; Nicolas Joly; Goran Jovanovic; Milija Jovanovic; Edward Lawton; Christopher McDonald; Jörg Schumacher; Christopher Waite; Nan Zhang

doi:10.1007/978-1-4939-2392-2_4

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

Martin Buck, Christoph Engl, Nicolas Joly, Goran Jovanovic, Milija Jovanovic, Edward Lawton, Christopher McDonald, Jörg Schumacher, Christopher Waite, Nan Zhang

Houston Methodist

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

1 Scopus citations

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 language	English (US)
Title of host publication	Bacterial Transcriptional Control
Subtitle of host publication	Methods and Protocols
Editors	Irina Artsimovitch, Thomas J. Santangelo
Publisher	Springer New York
Pages	53-79
Number of pages	27
Volume	1276
ISBN (Electronic)	9781493923922
ISBN (Print)	9781493923915
DOIs	https://doi.org/10.1007/978-1-4939-2392-2_4
State	Published - Feb 9 2015

Publication series

Name	Methods in Molecular Biology
Publisher	Springer Link

Keywords

AAA+ proteins
ATPase
Bacterial enhancer binding proteins
Open and closed promoter complexes
RNA polymerase
Transcription activation
σ

ASJC Scopus subject areas

Immunology and Microbiology(all)
Biochemistry, Genetics and Molecular Biology(all)

Access to Document

10.1007/978-1-4939-2392-2_4

Cite this

Buck, M., Engl, C., Joly, N., Jovanovic, G., Jovanovic, M., Lawton, E., McDonald, C., Schumacher, J., Waite, C., & 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 New York. https://doi.org/10.1007/978-1-4939-2392-2_4

In vitro and in vivo methodologies for studying the sigma 54-dependent transcription. / Buck, Martin; Engl, Christoph; Joly, Nicolas et al.
Bacterial Transcriptional Control: Methods and Protocols. ed. / Irina Artsimovitch; Thomas J. Santangelo. Vol. 1276 Springer New York, 2015. p. 53-79 (Methods in Molecular Biology).

Research output: Chapter in Book/Report/Conference proceeding › Chapter (peer-reviewed) › peer-review

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

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

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