Peptidyl-prolyl cis/trans isomerases (PPIases) are enzymes that assist in protein folding around proline-peptide bonds, and often possess chaperone activity. Staphylococcus aureus encodes three PPIases; PrsA, PpiB and Trigger factor (TF). Previous work by our group demonstrated a role for both PrsA and PpiB in S. aureus, however, TF remains largely unstudied. Here, we identify a role for TF in S. aureus biofilm formation, and demonstrate cooperation between TF and the cytoplasmic PPIase PpiB. Mutation of the tig gene (encoding TF) leads to reduced biofilm development in vitro but no significant attenuation of virulence in a mouse model of infection. To investigate if TF possesses chaperone activity, we analyzed the ability of a tig mutant to survive acid and basic stress. While there was no significant decrease in a tig mutant, a ppiB/tig double mutant exhibited a significant decrease in cell viability after acid and base challenge. We then demonstrate that a ppiB/tig double mutant has exacerbated phenotypes in vitro and in vivo when compared to either single mutant. Finally, in vivo immunoprecipitation of epitope tagged PpiB reveals that PpiB interacts with four times the number of proteins when TF is absent from the cell, suggesting it may be compensating for the loss of TF. Interestingly, the only proteins found to interact with TF are TF itself, FnBPB and the chaperone protein ClpB. Collectively, these results support the first phenotype for S. aureus TF and demonstrate a greater network of cooperation between chaperone proteins in Staphylococcus aureus.
IMPORTANCE S. aureus encodes a large number of virulence factors that aid the bacterium in survival and pathogenesis. These virulence factors have a wide variety of functions, however, they must all be properly secreted in order to be functional. Bacterial chaperone proteins often assist in secretion by trafficking proteins to secretion machinery or assisting in proper protein folding. Here, we report that the S. aureus chaperone Trigger factor (TF) contributes to biofilm formation and cooperates with the chaperone PpiB to regulate S. aureus virulence processes. These data highlight the first known role for TF in S. aureus, and suggest that S. aureus chaperone proteins may be involved in a greater regulatory network in the cell.