Hydrogen Sulfide and Reactive Sulfur Species Impact Proteome <i>S</i>‑Sulfhydration and Global Virulence Regulation in <i>Staphylococcus aureus</i> Hui Peng Yixiang Zhang Lauren D. Palmer Thomas E. Kehl-Fie Eric P. Skaar Jonathan C. Trinidad David P. Giedroc 10.1021/acsinfecdis.7b00090.s002 https://acs.figshare.com/articles/dataset/Hydrogen_Sulfide_and_Reactive_Sulfur_Species_Impact_Proteome_i_S_i_Sulfhydration_and_Global_Virulence_Regulation_in_i_Staphylococcus_aureus_i_/5382367 Hydrogen sulfide (H<sub>2</sub>S) is thought to protect bacteria from oxidative stress, but a comprehensive understanding of its function in bacteria is largely unexplored. In this study, we show that the human pathogen <i>Staphylococcus aureus</i> (<i>S. aureus</i>) harbors significant effector molecules of H<sub>2</sub>S signaling, reactive sulfur species (RSS), as low molecular weight persulfides of bacillithiol, coenzyme A, and cysteine, and significant inorganic polysulfide species. We find that proteome <i>S</i>-sulfhydration, a post-translational modification (PTM) in H<sub>2</sub>S signaling, is widespread in <i>S. aureus</i>. RSS levels modulate the expression of secreted virulence factors and the cytotoxicity of the secretome, consistent with an <i>S</i>-sulfhydration-dependent inhibition of DNA binding by MgrA, a global virulence regulator. Two previously uncharacterized thioredoxin-like proteins, denoted TrxP and TrxQ, are <i>S</i>-sulfhydrated in sulfide-stressed cells and are capable of reducing protein hydrodisulfides, suggesting that this PTM is potentially regulatory in <i>S. aureus</i>. In conclusion, our results reveal that <i>S. aureus</i> harbors a pool of proteome- and metabolite-derived RSS capable of impacting protein activities and gene regulation and that H<sub>2</sub>S signaling can be sensed by global regulators to affect the expression of virulence factors. 2017-08-29 00:00:00 . aureu S . aureus virulence factors bacteria PTM uncharacterized thioredoxin-like proteins expression DNA H 2 S Global Virulence Regulation regulator Staphylococcus aureus Hydrogen sulfide impacting protein activities S . aureus harbors pathogen Staphylococcus aureus proteome reactive sulfur species RSS