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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.

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