title: SalB inactivation modulates culture supernatant exoproteins and affects autolysis and viability in Enterococcus faecalis OG1RF.
abstract: The culture supernatant fraction of an Enterococcus faecalis gelE mutant of strain OG1RF contained elevated levels of the secreted antigen SalB. Using differential fluorescence gel electrophoresis (DIGE) the salB mutant was shown to possess a unique complement of exoproteins. Differentially abundant exoproteins were identified using matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Stress-related proteins including DnaK, Dps family protein, SOD, and NADH peroxidase were present in greater quantity in the OG1RF salB mutant culture supernatant. Moreover, several proteins involved in cell wall synthesis and cell division, including d-Ala-d-Lac ligase and EzrA, were present in reduced quantity in OG1RF salB relative to the parent strain. The salB mutant displayed reduced viability and anomalous cell division, and these phenotypes were exacerbated in a gelE salB double mutant. An epistatic relationship between gelE and salB was not identified with respect to increased autolysis and cell morphological changes observed in the salB mutant. SalB was purified as a six-histidine-tagged protein to investigate peptidoglycan hydrolytic activity; however, activity was not evident. High-pressure liquid chromatography (HPLC) analysis of reduced muropeptides from peptidoglycan digested with mutanolysin revealed that the salB mutant and OG1RF were indistinguishable.
authors.affiliation: Institute of Integrative Biology, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Jayendra
authors.initials: J
authors.lastname: Shankar
authors.forename: Rachel G
authors.initials: RG
authors.lastname: Walker
authors.forename: Mark C
authors.initials: MC
authors.lastname: Wilkinson
authors.forename: Deborah
authors.initials: D
authors.lastname: Ward
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: Horsburgh
mesh_terms: D000942:Antigens, Bacterial; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001433:Bacteriolysis; D003001:Cloning, Molecular; D013293:Enterococcus faecalis / Q000145:classification* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D050296:Microbial Viability; D009154:Mutation; D010457:Peptidoglycan / Q000235:genetics / Q000378:metabolism; D020543:Proteome / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013312:Stress, Physiological
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D010457:Peptidoglycan; D020543:Proteome