title: The chaperone ClpX stimulates expression of Staphylococcus aureus protein A by Rot dependent and independent pathways.
abstract: The Clp ATPases (Hsp100) constitute a family of closely related proteins that have protein reactivating and remodelling activities typical of molecular chaperones. In Staphylococcus aureus the ClpX chaperone is essential for virulence and for transcription of spa encoding Protein A. The present study was undertaken to elucidate the mechanism by which ClpX stimulates expression of Protein A. For this purpose, we prepared antibodies directed against Rot, an activator of spa transcription, and demonstrated that cells devoid of ClpX contain three-fold less Rot than wild-type cells. By varying Rot expression from an inducible promoter we showed that expression of Protein A requires a threshold level of Rot. In the absence of ClpX the Rot content is reduced below this threshold level, hence, explaining the substantially reduced Protein A expression in the clpX mutant. Experiments addressed at pinpointing the role of ClpX in Rot synthesis revealed that ClpX is required for translation of Rot. Interestingly, translation of the spa mRNA was, like the rot mRNA, enhanced by ClpX. These data demonstrate that ClpX performs dual roles in regulating Protein A expression, as ClpX stimulates transcription of spa by enhancing translation of Rot, and that ClpX additionally is required for full translation of the spa mRNA. The current findings emphasize that ClpX has a central role in fine-tuning virulence regulation in S. aureus.
authors.affiliation: Department of Veterinary Disease Biology, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark.
authors.forename: Lotte
authors.initials: L
authors.lastname: Jelsbak
authors.forename: Hanne
authors.initials: H
authors.lastname: Ingmer
authors.forename: Lukás
authors.initials: L
authors.lastname: Valihrach
authors.forename: Marianne Thorup
authors.initials: MT
authors.lastname: Cohn
authors.forename: Mie H G
authors.initials: MH
authors.lastname: Christiansen
authors.forename: Birgitte H
authors.initials: BH
authors.lastname: Kallipolitis
authors.forename: Dorte
authors.initials: D
authors.lastname: Frees
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D013205:Staphylococcal Protein A / Q000235:genetics* / Q000378:metabolism; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D018832:Molecular Chaperones; D012097:Repressor Proteins; D013205:Staphylococcal Protein A; C408643:rot protein, Staphylococcus aureus; D000251:Adenosine Triphosphatases