Promoter discrimination by the related transcriptional activators MarA and SoxS: differential regulation by differential binding

Mol Microbiol. 2000 Feb;35(3):623-34. doi: 10.1046/j.1365-2958.2000.01732.x.

Abstract

MarA and SoxS are closely related proteins ( approximately 45% identical) that transcriptionally activate a common set of unlinked genes, resulting in multiple antibiotic and superoxide resistance in Escherichia coli. Both proteins bind as monomers to a 20 bp degenerate asymmetric recognition sequence, the 'marbox', located upstream of the promoter. However, the proteins differ widely in the extents to which they activate particular promoters, with the consequence that overexpression of SoxS leads to greater superoxide resistance than does overexpression of MarA. This 'discrimination' between activators by promoters was demonstrated in vivo, using promoters fused to lacZ, and in vitro, using purified RNA polymerase, promoter DNA and MarA or SoxS. The marbox was found to be a critical element in discrimination by in vivo and in vitro assays of hybrid promoters containing the marbox from one gene and the core promoter from another. Furthermore, by sequential mutation of its marbox, a promoter that discriminated 35-fold in favour of SoxS was converted into one that did not discriminate. The relative activation of a promoter by MarA or SoxS was paralleled by the relative binding of the two activators to the promoter's marbox as assayed by band shift experiments. Thus, differential recognition of closely related marbox sequences by the closely related activators is the primary basis for promoter discrimination. Discrimination enables the cell to customize its response to the stresses that trigger synthesis of the activators.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / drug effects
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Base Sequence
  • Binding Sites
  • DNA-Binding Proteins / drug effects
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Drug Resistance, Microbial
  • Escherichia coli / drug effects
  • Escherichia coli / physiology
  • Escherichia coli Proteins*
  • Gene Expression Regulation, Bacterial*
  • Isopropyl Thiogalactoside / pharmacology
  • Methylphenazonium Methosulfate / pharmacology
  • Microbial Sensitivity Tests
  • Molecular Sequence Data
  • Nalidixic Acid / pharmacology
  • Norfloxacin / pharmacology
  • Promoter Regions, Genetic*
  • Substrate Specificity
  • Superoxides / metabolism
  • Superoxides / pharmacology
  • Trans-Activators / genetics
  • Trans-Activators / metabolism
  • Transcription Factors / drug effects
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Vitamin K / pharmacology

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • DNA-Binding Proteins
  • Escherichia coli Proteins
  • MarA protein, E coli
  • Rob protein, E coli
  • Trans-Activators
  • Transcription Factors
  • Superoxides
  • Vitamin K
  • SoxS protein, E coli
  • Methylphenazonium Methosulfate
  • Isopropyl Thiogalactoside
  • Nalidixic Acid
  • Norfloxacin