The antivirulent Staphylococcal sRNA SprC regulates CzrB efflux pump to adapt its response to zinc toxicity

RNA. 2024 Oct 16;30(11):1451-1464. doi: 10.1261/rna.080122.124.

Abstract

Bacterial regulatory RNAs (sRNAs) are important players to control gene expression. In Staphylococcus aureus, SprC is an antivirulent trans-acting sRNA known to base-pair with the major autolysin atl mRNA, preventing its translation. Using MS2-affinity purification coupled with RNA sequencing, we looked for its sRNA-RNA interactome and identified 14 novel mRNA targets. In vitro biochemical investigations revealed that SprC binds two of them, czrB and deoD, and uses a single accessible region to regulate its targets, including Atl translation. Unlike Atl regulation, the characterization of the SprC-czrB interaction pinpointed a destabilization of the czrAB cotranscript, leading to a decrease of the mRNA level that impaired CzrB zinc efflux pump expression. On a physiological standpoint, we showed that SprC expression is detrimental to combat against zinc toxicity. In addition, phagocyctosis assays revealed a significant, but moderate, increase of czrB mRNA levels in a sprC-deleted mutant, indicating a functional link between SprC and czrB upon internalization in macrophages, and suggesting a role in resistance to both oxidative and zinc bursts. Altogether, our data uncover a novel pathway in which SprC is implicated, highlighting the multiple strategies used by S. aureus to balance virulence using an RNA regulator.

Keywords: SprC; Staphylococcus aureus; czrB; sRNA; zinc.

MeSH terms

  • Bacterial Proteins* / genetics
  • Bacterial Proteins* / metabolism
  • Gene Expression Regulation, Bacterial*
  • Macrophages / drug effects
  • Macrophages / metabolism
  • Macrophages / microbiology
  • RNA, Bacterial* / genetics
  • RNA, Bacterial* / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Small Untranslated / genetics
  • RNA, Small Untranslated / metabolism
  • Staphylococcus aureus* / genetics
  • Staphylococcus aureus* / metabolism
  • Staphylococcus aureus* / pathogenicity
  • Virulence / genetics
  • Zinc* / metabolism
  • Zinc* / toxicity

Substances

  • Zinc
  • Bacterial Proteins
  • RNA, Bacterial
  • RNA, Messenger
  • RNA, Small Untranslated