In vitro evidence that RNA polymerase acetylation and acetyl Phosphate-Dependent CpxR phosphorylation affect CpxP transcription regulation

Bruno P. Lima, Christopher W. Lennon, Wilma Ross, Richard L. Gourse, Alan J. Wolfe

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

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 φ 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.

Original languageEnglish (US)
Article numberfnw011
JournalFEMS Microbiology Letters
Volume363
Issue number5
DOIs
StatePublished - Feb 8 2016

Bibliographical note

Funding Information:
We thank members of theWolfe and Gourse laboratories for critical discussion of the work presented here. This work was supported by NIH grant GM066130 and Loyola University Chicago Potts Foundation award LU11200 awarded to AJW and NIH GM37048 to RLG.

Publisher Copyright:
© FEMS 2016.

Keywords

  • Acetyl-phosphate
  • CpxAR
  • CpxP
  • In vitro transcription
  • Protein acetylation
  • RNA polymerase

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