title: Salmonella enterica serovar Typhimurium RamA, intracellular oxidative stress response, and bacterial virulence.
abstract: Escherichia coli and Salmonella enterica serovar Typhimurium have evolved genetic systems, such as the soxR/S and marA regulons, to detoxify reactive oxygen species, like superoxide, which are formed as by-products of metabolism. Superoxide also serves as a microbicidal effector mechanism of the host's phagocytes. Here, we investigate whether regulatory genes other than soxR/S and marA are active in response to oxidative stress in Salmonella and may function as virulence determinants. We identified a bacterial gene, which was designated ramA (342 bp) and mapped at 13.1 min on the Salmonella chromosome, that, when overexpressed on a plasmid in E. coli or Salmonella, confers a pleiotropic phenotype characterized by increased resistance to the redox-cycling agent menadione and to multiple unrelated antibiotics. The ramA gene is present in Salmonella serovars but is absent in E. coli. The gene product displays 37 to 52% homology to the transcriptional activators soxR/S and marA and 80 to 100% identity to a multidrug resistance gene in Klebsiella pneumoniae and Salmonella enterica serovar Paratyphi A. Although a ramA soxR/S double null mutant is highly susceptible to intracellular superoxide generated by menadione and displays decreased Mn-superoxide dismutase activity, intracellular survival of this mutant within macrophage-like RAW 264.7 cells and in vivo replication in the spleens in Ityr mice are not affected. We concluded that despite its role in the protective response of the bacteria to oxidative stress in vitro, the newly identified ramA gene, together with soxR/S, does not play a role in initial replication of Salmonella in the organs of mice.
authors.affiliation: Department of Infectious Diseases, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
authors.forename: Tahar
authors.initials: T
authors.lastname: van der Straaten
authors.forename: Laurence
authors.initials: L
authors.lastname: Zulianello
authors.forename: Angela
authors.initials: A
authors.lastname: van Diepen
authors.forename: Donald L
authors.initials: DL
authors.lastname: Granger
authors.forename: Riny
authors.initials: R
authors.lastname: Janssen
authors.forename: Jaap T
authors.initials: JT
authors.lastname: van Dissel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D051777:AraC Transcription Factor; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics; D008264:Macrophages / Q000382:microbiology; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D018384:Oxidative Stress; D018085:Regulon; D012097:Repressor Proteins; D012486:Salmonella typhimurium / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics; D014774:Virulence
chemical_list: D051777:AraC Transcription Factor; C497371:AraC protein, E coli; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C080002:MarA protein, E coli; C579436:RamA protein, Salmonella enterica; D012097:Repressor Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D013481:Superoxides; C070724:SoxR protein, Bacteria; D013482:Superoxide Dismutase