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

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

1 Scopus citations


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 (US)
Title of host publicationBacterial Transcriptional Control
Subtitle of host publicationMethods and Protocols
EditorsIrina Artsimovitch, Thomas J. Santangelo
PublisherSpringer New York
Number of pages27
ISBN (Electronic)9781493923922
ISBN (Print)9781493923915
StatePublished - Feb 9 2015

Publication series

NameMethods in Molecular Biology
PublisherSpringer Link


  • 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)


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