title: In vitro and in vivo methodologies for studying the Sigma 54-dependent transcription.
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.
authors: Department of Life Sciences, Imperial College London, South Kensington Campus, London, SW7 2AZ, UK, m.buck@imperial.ac.uk.
authors: Martin
authors: M
authors: Buck
authors: Christoph
authors: C
authors: Engl
authors: Nicolas
authors: N
authors: Joly
authors: Goran
authors: G
authors: Jovanovic
authors: Milija
authors: M
authors: Jovanovic
authors: Edward
authors: E
authors: Lawton
authors: Christopher
authors: C
authors: McDonald
authors: Jörg
authors: J
authors: Schumacher
authors: Christopher
authors: C
authors: Waite
authors: Nan
authors: N
authors: Zhang
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000302:isolation & purification; D001483:Base Sequence; D002855:Chromatography, Thin Layer; D018983:DNA Footprinting / Q000379:methods; D004268:DNA-Binding Proteins / Q000302:isolation & purification; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D066298:In Vitro Techniques; D008958:Models, Molecular; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D051877:RNA Polymerase Sigma 54 / Q000737:chemistry / Q000378:metabolism*; D015534:Trans-Activators / Q000302:isolation & purification; D014157:Transcription Factors / Q000302:isolation & purification; D014158:Transcription, Genetic / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C085856:HrpR protein, Pseudomonas syringae; C098954:PspF protein, E coli; D015534:Trans-Activators; D014157:Transcription Factors; C497751:rpoN protein, E coli; C060789:hrpS protein, Pseudomonas syringae; D051877:RNA Polymerase Sigma 54; D000251:Adenosine Triphosphatases