title: In vitro evidence that RNA Polymerase acetylation and acetyl phosphate-dependent CpxR phosphorylation affect cpxP transcription regulation.
abstract: The central metabolite acetyl phosphate (acP) has long been proposed to influence transcription regulation by directly transferring its phosphoryl group to a number of response regulators in many bacterial species. Here, we provide in vitro evidence for this proposition and demonstrate, using an in vitro transcription system, that acP-dependent phosphorylation of aspartate 51 of CpxR induces transcription of one of its regulon members in E. coli, cpxP. We also used this in vitro transcription system to extend our previously reported in vivo data that hypothesized that acetylation of RNA polymerase (RNAP) influences acP-dependent cpxP transcription, using glutamine as a genetic mimic for acetylated arginine 291 of the carboxy-terminal domain of RNAP α subunit. The data we present here lend strong support to the hypothesis that acP has a direct effect on transcription regulation in E. coli via phosphorylation of CpxR, and that RNAP acetylation can modulate this response.
authors.affiliation: Department of Microbiology and Immunology, Stritch School of Medicine, Health Sciences Division, Loyola University Chicago, Maywood, IL 60153, USA.
authors.forename: Bruno P
authors.initials: BP
authors.lastname: Lima
authors.affiliation: Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Christopher W
authors.initials: CW
authors.lastname: Lennon
authors.affiliation: Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Wilma
authors.initials: W
authors.lastname: Ross
authors.affiliation: Department of Bacteriology, University of Wisconsin-Madison, Madison, WI 53706, USA.
authors.forename: Richard L
authors.initials: RL
authors.lastname: Gourse
authors.affiliation: Department of Microbiology and Immunology, Stritch School of Medicine, Health Sciences Division, Loyola University Chicago, Maywood, IL 60153, USA awolfe@luc.edu.
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Wolfe
mesh_terms: D000107:Acetylation; D001426:Bacterial Proteins / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D010755:Organophosphates / Q000737:chemistry*; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000235:genetics; D011499:Protein Processing, Post-Translational; D018085:Regulon
chemical_list: D001426:Bacterial Proteins; C491730:CpxP protein, E coli; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D010755:Organophosphates; C085504:CpxR protein, Bacteria; C011632:acetyl phosphate; D012321:DNA-Directed RNA Polymerases; C113275:RNA polymerase alpha subunit
keywords: CpxAR; In vitro transcription; RNA polymerase; acetyl-phosphate; cpxP; protein acetylation