ClpC affects the intracellular survival capacity of Staphylococcus aureus in non-professional phagocytic cells

Sci Rep. 2019 Nov 7;9(1):16267. doi: 10.1038/s41598-019-52731-3.

Abstract

Invasion and persistence of bacteria within host cells requires that they adapt to life in an intracellular environment. This adaptation induces bacterial stress through events such as phagocytosis and enhanced nutrient-restriction. During stress, bacteria synthesize a family of proteins known as heat shock proteins (HSPs) to facilitate adaptation and survival. Previously, we determined the Staphylococcus aureus HSP ClpC temporally alters bacterial metabolism and persistence. This led us to hypothesize that ClpC might alter intracellular survival. Inactivation of clpC in S. aureus strain DSM20231 significantly enhanced long-term intracellular survival in human epithelial (HaCaT) and endothelial (EA.hy926) cell lines, without markedly affecting adhesion or invasion. This phenotype was similar across a genetically diverse collection of S. aureus isolates, and was influenced by the toxin/antitoxin encoding locus mazEF. Importantly, MazEF alters mRNA synthesis and/or stability of S. aureus virulence determinants, indicating ClpC may act through the mRNA modulatory activity of MazEF. Transcriptional analyses of total RNAs isolated from intracellular DSM20231 and isogenic clpC mutant cells identified alterations in transcription of α-toxin (hla), protein A (spa), and RNAIII, consistent with the hypothesis that ClpC negatively affects the intracellular survival of S. aureus in non-professional phagocytic cells, via modulation of MazEF and Agr.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Adhesion
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Cytotoxicity, Immunologic
  • Heat-Shock Proteins / genetics*
  • Heat-Shock Proteins / metabolism
  • Host-Pathogen Interactions* / genetics
  • Host-Pathogen Interactions* / immunology
  • Humans
  • Microbial Viability / immunology
  • Mutation
  • Phagocytes / immunology*
  • Phagocytes / metabolism
  • Phagocytes / microbiology*
  • Staphylococcal Infections / genetics*
  • Staphylococcal Infections / immunology*
  • Staphylococcal Infections / microbiology
  • Staphylococcus aureus / physiology*
  • Transcriptional Activation
  • Virulence

Substances

  • Bacterial Proteins
  • ClpC protein, Bacteria
  • Heat-Shock Proteins