CodY regulates thiol peroxidase expression as part of the pneumococcal defense mechanism against H2 O2 stress

Barak Hajaj, Hasan Yesilkaya, Sulman Shafeeq, Xiangyun Zhi, Rachel Benisty, Shiran Tchalah, Oscar P. Kuipers, Nurith Porat

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Streptococcus pneumoniae is a facultative anaerobic pathogen. Although it maintains fermentative metabolism, during aerobic growth pneumococci produce high levels of H2 O2 , which can have adverse effects on cell viability and DNA, and influence pneumococcal interaction with its host. The pneumococcus is unusual in its dealing with toxic reactive oxygen species (ROS) in that it neither has catalase nor the global regulators of peroxide stress resistance. Previously, we identified pneumococcal thiol peroxidase (TpxD) as the key enzyme for enzymatic removal of H2 O2 , and showed that TpxD synthesis is up-regulated upon exposure to H2 O2 . This study aimed to reveal the mechanism controlling TpxD expression under H2 O2 stress. We hypothesize that H2 O2 activates a transcription factor which in turn up-regulates tpxD expression. Microarray analysis revealed a pneumococcal global transcriptional response to H2 O2 . Mutation of tpxD abolished H2 O2 -mediated response to high H2 O2 levels, signifying the need for an active TpxD under oxidative stress conditions. Bioinformatic tools, applied to search for a transcription factor modulating tpxD expression, pointed toward CodY as a potential candidate. Indeed, a putative 15-bp consensus CodY binding site was found in the proximal region of tpxD-coding sequence. Binding of CodY to this site was confirmed by EMSA, and genetic engineering techniques demonstrated that this site is essential for TpxD up-regulation under H2 O2 stress. Furthermore, tpxD expression was reduced in a ΔcodY mutant. These data indicate that CodY is an activator of tpxD expression, triggering its up-regulation under H2 O2 stress. In addition we show that H2 O2 specifically oxidizes the 2 CodY cysteines. This oxidation may trigger a conformational change in CodY, resulting in enhanced binding to DNA. A schematic model illustrating the contribution of TpxD and CodY to pneumococcal global transcriptional response to H2 O2 is proposed.

Original languageEnglish
Article number210
JournalFrontiers in Cellular and Infection Microbiology
Volume7
Issue numberMAY
DOIs
StatePublished - 24 May 2017
Externally publishedYes

Keywords

  • CodY
  • Global gene regulation
  • H O
  • Streptococcus pneumoniae
  • TpxD

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Microbiology (medical)
  • Infectious Diseases

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