title: Characterization of the sortase repertoire in Bacillus anthracis.
abstract: LPXTG proteins, present in most if not all Gram-positive bacteria, are known to be anchored by sortases to the bacterial peptidoglycan. More than one sortase gene is often encoded in a bacterial species, and each sortase is supposed to specifically anchor given LPXTG proteins, depending of the sequence of the C-terminal cell wall sorting signal (cwss), bearing an LPXTG motif or another recognition sequence. B. anthracis possesses three sortase genes. B. anthracis sortase deleted mutant strains are not affected in their virulence. To determine the sortase repertoires, we developed a genetic screen using the property of the gamma phage to lyse bacteria only when its receptor, GamR, an LPXTG protein, is exposed at the surface. We identified 10 proteins that contain a cell wall sorting signal and are covalently anchored to the peptidoglycan. Some chimeric proteins yielded phage lysis in all sortase mutant strains, suggesting that cwss proteins remained surface accessible in absence of their anchoring sortase, probably as a consequence of membrane localization of yet uncleaved precursor proteins. For definite assignment of the sortase repertoires, we consequently relied on a complementary test, using a biochemical approach, namely immunoblot experiments. The sortase anchoring nine of these proteins has thus been determined. The absence of virulence defect of the sortase mutants could be a consequence of the membrane localization of the cwss proteins.
authors.affiliation: Institut Pasteur, Unités Toxines et Pathogénie Bactérienne, Pathogénèse des Toxi-infections Bactériennes, Département de Microbiologie, Paris, France.
authors.forename: Willy
authors.initials: W
authors.lastname: Aucher
authors.forename: Sophie
authors.initials: S
authors.lastname: Davison
authors.forename: Agnès
authors.initials: A
authors.lastname: Fouet
mesh_terms: D020816:Amino Acid Motifs; D019881:Aminoacyltransferases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000881:Anthrax / Q000378:metabolism*; D001408:Bacillus anthracis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D010582:Bacteriophage lambda / Q000235:genetics / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D050506:Mutant Chimeric Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010446:Peptide Fragments / Q000276:immunology; D010457:Peptidoglycan / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000502:physiology*; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D050506:Mutant Chimeric Proteins; D010446:Peptide Fragments; D010457:Peptidoglycan; D021382:Protein Sorting Signals; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases
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