title: Diversity of two-component systems: insights into the signal transduction mechanism by the  Staphylococcus aureus two-component system GraSR.
abstract: The response to cationic antimicrobial peptides (CAMPs) in Staphylococcus aureus relies on a two-component system (TCS), GraSR, an auxiliary protein GraX and an ATP-binding cassette (ABC) transporter, VraF/G. To understand the signal transduction mechanism by GraSR, we investigated the kinase activity of the cytoplasmic domain of histidine kinase GraS and the interaction with its cognate response regulator GraR. We also investigated interactions among the auxiliary protein GraX, GraS/R and the ATPase protein of the ABC transporter, VraF. We found that GraS lacks autophosphorylation activity, unlike a similar histidine kinase, BceS, of Bacillus subtilis. In addition, the interaction between GraS and GraR is very weak in comparison to the stronger interaction observed between BceS and its conjugated response regulator, BceR, suggesting that CAMP signaling may not flow directly from GraS to GraR. We found that the auxiliary protein GraX interacts with VraF and GraR, and requires the histidine phosphotransfer and dimerization domain of GraS to interact with this protein. Further, VraF requires the GraS region that connects the membrane-bound domain with the cytoplasmic domain of this protein for interaction with GraS. The interactions of GraX with GraS/R and VraF indicate that GraX may serve as a scaffold to bring these proteins in close proximity to GraS, plausibly to facilitate activation of GraS to ultimately transduce the signal to GraR.
authors.affiliation: Departments of Biology, York University, Toronto, Toronto, ON, M3J 1P3, Canada.
authors.forename: Uzma
authors.initials: U
authors.lastname: Muzamal
authors.affiliation: Department of Chemistry, York University, Toronto, Toronto, ON, M3J 1P3, Canada.
authors.forename: Daniel
authors.initials: D
authors.lastname: Gomez
authors.affiliation: Departments of Biology, York University, Toronto, Toronto, ON, M3J 1P3, Canada.
authors.forename: Fenika
authors.initials: F
authors.lastname: Kapadia
authors.affiliation: Departments of Biology, York University, Toronto, Toronto, ON, M3J 1P3, Canada ; Department of Chemistry, York University, Toronto, Toronto, ON, M3J 1P3, Canada.
authors.forename: Dasantila
authors.initials: D
authors.lastname: Golemi-Kotra
keywords: GraSR; Staphylococcus aureus; Two-component systems; cationic antimicrobial peptides; histidine kinases; response regulators