title: The srhSR gene pair from Staphylococcus aureus: genomic and proteomic approaches to the identification and characterization of gene function.
abstract: Systematic analysis of the entire two-component signal transduction system (TCSTS) gene complement of Staphylococcus aureus revealed the presence of a putative TCSTS (designated SrhSR) which shares considerable homology with the ResDE His-Asp phospho-relay pair of Bacillus subtilis. Disruption of the srhSR gene pair resulted in a dramatic reduction in growth of the srhSR mutant, when cultured under anaerobic conditions, and a 3-log attenuation in growth when analyzed in the murine pyelonephritis model. To further understand the role of SrhSR, differential display two-dimensional gel electrophoresis was used to analyze the cell-free extracts derived from the srhSR mutant and the corresponding wild type. Proteins shown to be differentially regulated were identified by mass spectrometry in combination with protein database searching. An srhSR deletion led to changes in the expression of proteins involved in energy metabolism and other metabolic processes including arginine catabolism, xanthine catabolism, and cell morphology. The impaired growth of the mutant under anaerobic conditions and the dramatic changes in proteins involved in energy metabolism shed light on the mechanisms used by S. aureus to grow anaerobically and indicate that the staphylococcal SrhSR system plays an important role in the regulation of energy transduction in response to changes in oxygen availability. The combination of proteomics, bio-informatics, and microbial genetics employed here represents a powerful set of techniques which can be applied to the study of bacterial gene function.
authors.affiliation: Anti-infectives Research, GlaxoSmithKline Pharmaceuticals Research and Development, Collegeville, Pennsylvania 19426, USA. John_Throup-1@sbphrd.com
authors.forename: J P
authors.initials: JP
authors.lastname: Throup
authors.forename: F
authors.initials: F
authors.lastname: Zappacosta
authors.forename: R D
authors.initials: RD
authors.lastname: Lunsford
authors.forename: R S
authors.initials: RS
authors.lastname: Annan
authors.forename: S A
authors.initials: SA
authors.lastname: Carr
authors.forename: J T
authors.initials: JT
authors.lastname: Lonsdale
authors.forename: A P
authors.initials: AP
authors.lastname: Bryant
authors.forename: D
authors.initials: D
authors.lastname: McDevitt
authors.forename: M
authors.initials: M
authors.lastname: Rosenberg
authors.forename: M K
authors.initials: MK
authors.lastname: Burnham
mesh_terms: D000595:Amino Acid Sequence; D001224:Aspartic Acid; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D019992:Databases as Topic; D017353:Gene Deletion; D005798:Genes, Bacterial; D023281:Genomics; D006639:Histidine; D000071677:Histidine Kinase; D007700:Kinetics; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D019151:Peptide Library; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics; D020543:Proteome; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction / Q000235:genetics; D013211:Staphylococcus aureus / Q000235:genetics* / Q000254:growth & development
chemical_list: D001426:Bacterial Proteins; D019151:Peptide Library; D020543:Proteome; C435497:SrhS protein, Staphylococcus aureus; D001224:Aspartic Acid; D006639:Histidine; D011494:Protein Kinases; D000071677:Histidine Kinase