The dual role of DksA protein in the regulation of Escherichia coli pArgX promoter

Nucleic Acids Res. 2016 Dec 1;44(21):10316-10325. doi: 10.1093/nar/gkw912. Epub 2016 Oct 7.

Abstract

Gene expression regulation by the stringent response effector, ppGpp, is facilitated by DksA protein; however DksA and ppGpp can play independent roles in transcription. In Escherichia coli, the pArgX promoter which initiates the transcription of four tRNA genes was shown to be inhibited by ppGpp. Our studies on the role of DksA in pArgX regulation revealed that it can stimulate transcription by increasing the binding of RNA polymerase to the promoter and the productive transcription complex formation. However, when DksA is present together with ppGpp a severe down-regulation of promoter activity is observed. Our results indicate that DksA facilitates the effects of ppGpp to drive formation of inactive dead-end complexes formed by RNA polymerase at the ArgX promoter. In vivo, ppGpp-mediated regulation of pArgX transcription is dependent on DksA activity. The potential mechanisms of opposing pArgX regulation by ppGpp and DksA are discussed. pArgX is the first reported example of the promoter stimulated by DksA and inhibited by ppGpp in vitro when an overall inhibition occurs in the presence of both regulators. A dual role is thus proposed for DksA in the regulation of the pArgX promoter activity.

MeSH terms

  • DNA, Bacterial / genetics
  • DNA, Bacterial / metabolism
  • DNA-Directed RNA Polymerases / metabolism
  • Escherichia coli / genetics*
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / genetics*
  • Escherichia coli Proteins / metabolism*
  • Gene Expression Regulation, Bacterial*
  • Promoter Regions, Genetic*
  • Protein Binding
  • Transcription Initiation, Genetic
  • Transcription, Genetic

Substances

  • DNA, Bacterial
  • Escherichia coli Proteins
  • dksA protein, E coli
  • DNA-Directed RNA Polymerases