Genetic, enzymatic, and pathogenic studies of the iron superoxide dismutase of Campylobacter jejuni

E. C. Pesci, D. L. Cottle, C. L. Pickett

Research output: Contribution to journalArticlepeer-review

117 Scopus citations

Abstract

Campylobacter jejuni is a microaerobic bacterium that produces an acute, self-limiting, watery or bloody diarrhea in humans. Little is known about how C. jejuni causes disease or even what specific capabilities it requires for survival in vivo. The enzyme, superoxide dismutase (SOD), which catalyzes the breakdown of superoxide radicals to hydrogen peroxide and dioxygen is one of the bacterial cell's major defense mechanisms against oxidative damage. A PCR-based search for sod genes in C. jejuni 81-176 revealed that this bacterium contained at least one sod gene. We cloned and sequenced a sod gene from 81-176 and determined that its predicted protein product was most similar to that of FeSODs (sodB genes). Transcriptional analysis indicated that this gene is monocistronic and may be transcribed from a σ70-like promoter. Nondenaturing polyacrylamide gels stained to reveal SOD activities, accompanied by inhibition studies, demonstrated that C. jejuni produces five electrophoretically distinct bands of SOD activity, all of which appeared to be FcSODs. Analysis of an 81-176 sodB strain revealed that all of these FeSOD activities may be products of the one sodB gene that we cloned. The expression and enzymatic activity of the respective sodB and FeSOD produced by both C. jejuni and Helicobacter pylori were examined in Escherichia coli. Both genes were expressed in E. coli, and the proteins produced were enzymatically active. Finally, the ability of the 81-176 sodB strain to survive INT-407 cell invasion was found to be significantly decreased (12- fold) compared with that of the parent, suggesting a potential role for SodB in C. jejuni intracellular survival.

Original languageEnglish
Pages (from-to)2687-2694
Number of pages8
JournalInfection and Immunity
Volume62
Issue number7
StatePublished - 1994

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

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