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

Authors
  • Buck, Martin
  • Engl, Christoph
  • Joly, Nicolas
  • Jovanovic, Goran
  • Jovanovic, Milija
  • Lawton, Edward
  • McDonald, Christopher
  • Schumacher, Jörg
  • Waite, Christopher
  • Zhang, Nan
Type
Published Article
Journal
Methods in Molecular Biology
Publication Date
Jan 01, 2015
Volume
1276
Pages
53–79
Identifiers
DOI: 10.1007/978-1-4939-2392-2_4
PMID: 25665558
Source
Medline
License
Unknown

Abstract

Here we describe approaches and methods to assaying in vitro the major variant bacterial sigma factor, Sigma 54 (σ(54)), 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 σ(54)-dependent gene expression.

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