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PMID: 11914350 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Differential recognition of surface proteins in Streptococcus pyogenes by two sortase gene homologs.

Journal of bacteriology ·Vol. 184 ·No. 8 ·2002-04-00 ·Pages 2181-91

Barnett TC, Scott JR

Abstract

The interaction of Streptococcus pyogenes (group A streptococcus [GAS]) with its human host requires several surface proteins. In this study, we isolated mutations in a gene required for the surface localization of protein F by transposon mutagenesis of the M6 strain JRS4. This gene (srtA) encodes a protein homologous to Staphylococcus aureus sortase, which covalently links proteins containing an LPXTG motif to the cell wall. The GAS srtA mutant was defective in anchoring the LPXTG-containing proteins M6, protein F, ScpA, and GRAB to the cell surface. This phenotype was complemented when a wild-type srtA gene was provided in trans. The surface localization of T6, however, was unaffected by the srtA mutation. The M1 genome sequence contains a second open reading frame with a motif characteristic of sortase proteins. Inactivation of this gene (designated srtB) in strain JRS4 affected the surface localization of T6 but not M6, protein F, ScpA, or GRAB. This phenotype was complemented by srtB in trans. An srtA probe hybridized with DNA from all GAS strains tested (M types 1, 3, 4, 5, 6, 18, 22, and 50 and nontypeable strain 64/14) and from streptococcal groups C and G, while srtB hybridized with DNA from only a few GAS strains. We conclude that srtA and srtB encode sortase enzymes required for anchoring different subsets of proteins to the cell wall. It seems likely that the multiple sortase homologs in the genomes of other gram-positive bacteria have a similar substrate-specific role.

MeSH Terms
Amino Acid Sequence Aminoacyltransferases/chemistry,genetics,physiology Bacterial Proteins Cell Wall/chemistry Cysteine Endopeptidases DNA Transposable Elements Molecular Sequence Data Mutation Streptococcus pyogenes/enzymology,genetics
Chemicals
Bacterial Proteins DNA Transposable Elements Aminoacyltransferases sortase A Cysteine Endopeptidases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Barnett Timothy C
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Scott June R
References (70)
70 references, click to expand
  1. Sortase-catalysed anchoring of surface proteins to the cell wall of Staphylococcus aureus.
    Mol Microbiol. 2001 Jun;40(5):1049-57 PMID: 11401711
  2. Unique regulation of SclB - a novel collagen-like surface protein of Streptococcus pyogenes.
    Mol Microbiol. 2001 Jun;40(6):1427-38 PMID: 11442840
  3. Generation and surface localization of intact M protein in Streptococcus pyogenes are dependent on sagA.
    Infect Immun. 2001 Nov;69(11):7029-38 PMID: 11598078
  4. Fba, a novel fibronectin-binding protein from Streptococcus pyogenes, promotes bacterial entry into epithelial cells, and the fba gene is positively transcribed under the Mga regulator.
    Mol Microbiol. 2001 Oct;42(1):75-86 PMID: 11679068
  5. Genomic localization of a T serotype locus to a recombinatorial zone encoding extracellular matrix-binding proteins in Streptococcus pyogenes.
    Infect Immun. 2002 Mar;70(3):1159-67 PMID: 11854196
  6. Peptidoglycan types of bacterial cell walls and their taxonomic implications.
    Bacteriol Rev. 1972 Dec;36(4):407-77 PMID: 4568761
  7. A role for trigger factor and an rgg-like regulator in the transcription, secretion and processing of the cysteine proteinase of Streptococcus pyogenes.
    EMBO J. 1998 Nov 2;17(21):6263-75 PMID: 9799235
  8. Serum opacity factor is a major fibronectin-binding protein and a virulence determinant of M type 2 Streptococcus pyogenes.
    Mol Microbiol. 1999 Apr;32(1):89-98 PMID: 10216862
  9. Protein GRAB of streptococcus pyogenes regulates proteolysis at the bacterial surface by binding alpha2-macroglobulin.
    J Biol Chem. 1999 May 28;274(22):15336-44 PMID: 10336419
  10. Staphylococcus aureus sortase, an enzyme that anchors surface proteins to the cell wall.
    Science. 1999 Jul 30;285(5428):760-3 PMID: 10427003
  11. Anchor structure of staphylococcal surface proteins. IV. Inhibitors of the cell wall sorting reaction.
    J Biol Chem. 1999 Aug 20;274(34):24316-20 PMID: 10446208
  12. Current knowledge of type-specific M antigens of group A streptococci.
    J Immunol. 1962 Sep;89:307-13 PMID: 14461914
  13. Environmental regulation of virulence in group A streptococci: transcription of the gene encoding M protein is stimulated by carbon dioxide.
    J Bacteriol. 1992 Sep;174(17):5693-701 PMID: 1512202
  14. Expression of protein F, the fibronectin-binding protein of Streptococcus pyogenes JRS4, in heterologous streptococcal and enterococcal strains promotes their adherence to respiratory epithelial cells.
    Infect Immun. 1992 Dec;60(12):5119-25 PMID: 1452345
  15. Identification of a lysin associated with a bacteriophage (A25) virulent for group A streptococci.
    J Bacteriol. 1981 Feb;145(2):696-703 PMID: 7007344
  16. Antiopsonic activity of fibrinogen bound to M protein on the surface of group A streptococci.
    J Clin Invest. 1982 Apr;69(4):1042-5 PMID: 7042754
  17. Cloning in Bacillus subtilis of an extremely thermostable alpha amylase: comparison with other cloned heatstable alpha amylases.
    Biochem Biophys Res Commun. 1984 Jul 18;122(1):175-83 PMID: 6331442
  18. Relationship of M protein genes in group A streptococci.
    Proc Natl Acad Sci U S A. 1985 Mar;82(6):1822-6 PMID: 3885219
  19. Production and biological properties of M-protein of Streptococcus equi.
    Res Vet Sci. 1985 Mar;38(2):184-8 PMID: 4001557
  20. Complete nucleotide sequence of type 6 M protein of the group A Streptococcus. Repetitive structure and membrane anchor.
    J Biol Chem. 1986 Feb 5;261(4):1677-86 PMID: 3511046
  21. Immunochemical localization and amino acid sequences of crossreactive epitopes within the group A streptococcal M6 protein.
    J Exp Med. 1986 Oct 1;164(4):1226-38 PMID: 2428914
  22. Conversion of an M- group A streptococcus to M+ by transfer of a plasmid containing an M6 gene.
    J Exp Med. 1986 Nov 1;164(5):1641-51 PMID: 3534131
  23. Biological and immunochemical identity of M protein on group G streptococci with M protein on group A streptococci.
    Infect Immun. 1987 Mar;55(3):502-6 PMID: 3546129
  24. Complete nucleotide sequence of the streptococcal C5a peptidase gene of Streptococcus pyogenes.
    J Biol Chem. 1990 Feb 25;265(6):3161-7 PMID: 2406246
  25. Sequence and structural characteristics of the trypsin-resistant T6 surface protein of group A streptococci.
    J Bacteriol. 1990 Jun;172(6):3310-7 PMID: 2188957
  26. Conservation of a hexapeptide sequence in the anchor region of surface proteins from gram-positive cocci.
    Mol Microbiol. 1990 Sep;4(9):1603-5 PMID: 2287281
  27. Characterization of the Lactococcus lactis lactose operon promoter: contribution of flanking sequences and LacR repressor to promoter activity.
    J Bacteriol. 1992 Apr;174(7):2273-80 PMID: 1372602
  28. Protein F, a fibronectin-binding protein, is an adhesin of the group A streptococcus Streptococcus pyogenes.
    Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6172-6 PMID: 1385871
  29. Group G streptococcal M protein exhibits structural features analogous to those of class I M protein of group A streptococci.
    Infect Immun. 1992 Sep;60(9):3689-96 PMID: 1500178
  30. Isolation and characterization of genetic expression and secretion signals from Enterococcus faecalis through the use of broad-host-range alpha-amylase probe vectors.
    Gene. 1992 Sep 1;118(1):21-30 PMID: 1511881
  31. Streptococcal C5a peptidase is a highly specific endopeptidase.
    Infect Immun. 1992 Dec;60(12):5219-23 PMID: 1452354
  32. Introduction of the emm6 gene into an emm-deleted strain of Streptococcus pyogenes restores its ability to resist phagocytosis.
    Res Microbiol. 1992 Jul-Aug;143(6):549-58 PMID: 1475516
  33. Role of M protein in pharyngeal colonization by group A streptococci in rats.
    Infect Immun. 1993 Jun;61(6):2277-83 PMID: 8500869
  34. Studies of the receptor for phage A25 in group A streptococci: the role of peptidoglycan in reversible adsorption.
    J Exp Med. 1977 Mar 1;145(3):578-93 PMID: 233903
  35. Nucleotide substitutions and small-scale insertion produce size and antigenic variation in group A streptococcal M1 protein.
    Mol Microbiol. 1993 May;8(5):981-91 PMID: 8355619
  36. Simultaneous production and purification of Bacillus subtilis alpha-amylase.
    Appl Biochem Biotechnol. 1993 Mar;38(3):269-76 PMID: 7690533
  37. Cell wall sorting signals in surface proteins of gram-positive bacteria.
    EMBO J. 1993 Dec;12(12):4803-11 PMID: 8223489
  38. Use of the Escherichia coli beta-glucuronidase (gusA) gene as a reporter gene for analyzing promoters in lactic acid bacteria.
    Appl Environ Microbiol. 1994 Feb;60(2):587-93 PMID: 8135517
  39. Protein F: an adhesin of Streptococcus pyogenes binds fibronectin via two distinct domains.
    Mol Microbiol. 1993 Dec;10(5):1049-55 PMID: 7934855
  40. Expression of the Arp protein, a member of the M protein family, is not sufficient to inhibit phagocytosis of Streptococcus pyogenes.
    Infect Immun. 1995 Jan;63(1):345-8 PMID: 7806375
  41. Proteolytic cleavage and cell wall anchoring at the LPXTG motif of surface proteins in gram-positive bacteria.
    Mol Microbiol. 1994 Oct;14(1):115-21 PMID: 7830549
  42. C5a peptidase alters clearance and trafficking of group A streptococci by infected mice.
    Infect Immun. 1996 Feb;64(2):503-10 PMID: 8550199
  43. CotM of Bacillus subtilis, a member of the alpha-crystallin family of stress proteins, is induced during development and participates in spore outer coat formation.
    J Bacteriol. 1997 Mar;179(6):1887-97 PMID: 9068633
  44. Cell wall anchoring of the Streptococcus pyogenes M6 protein in various lactic acid bacteria.
    J Bacteriol. 1997 May;179(9):3068-72 PMID: 9139932
  45. comYA, a gene similar to comGA of Bacillus subtilis, is essential for competence-factor-dependent DNA transformation in Streptococcus gordonii.
    J Bacteriol. 1997 May;179(10):3122-6 PMID: 9150204
  46. Intranasal immunization with C5a peptidase prevents nasopharyngeal colonization of mice by the group A Streptococcus.
    Infect Immun. 1997 Jun;65(6):2080-7 PMID: 9169735
  47. Affinity purification and characterization of a fibrinogen-binding protein complex which protects mice against lethal challenge with Streptococcus equi subsp. equi.
    Microbiology. 1998 Apr;144 ( Pt 4):993-1003 PMID: 9579073
  48. Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis.
    Mol Microbiol. 1998 May;28(3):449-61 PMID: 9632250
  49. Use of the lactococcal nisA promoter to regulate gene expression in gram-positive bacteria: comparison of induction level and promoter strength.
    Appl Environ Microbiol. 1998 Aug;64(8):2763-9 PMID: 9687428
  50. Molecular analysis of the role of the group A streptococcal cysteine protease, hyaluronic acid capsule, and M protein in a murine model of human invasive soft-tissue infection.
    J Clin Invest. 1998 Aug 1;102(3):550-60 PMID: 9691092
  51. Group A streptococcal isolate 64/14 expresses surface plasmin-binding structures in addition to Plr.
    Res Microbiol. 1997 Sep-Oct;148(7):559-72 PMID: 9765841
  52. Anchor structure of staphylococcal surface proteins. II. Cooh-terminal structure of muramidase and amidase-solubilized surface protein.
    J Biol Chem. 1998 Oct 30;273(44):29135-42 PMID: 9786922
  53. Purification and characterization of sortase, the transpeptidase that cleaves surface proteins of Staphylococcus aureus at the LPXTG motif.
    Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12424-9 PMID: 10535938
  54. Anchoring of surface proteins to the cell wall of Staphylococcus aureus. Sortase catalyzed in vitro transpeptidation reaction using LPXTG peptide and NH(2)-Gly(3) substrates.
    J Biol Chem. 2000 Mar 31;275(13):9876-81 PMID: 10734144
  55. Sortase, a universal target for therapeutic agents against gram-positive bacteria?
    Proc Natl Acad Sci U S A. 2000 May 9;97(10):5013-5 PMID: 10805759
  56. Staphylococcus aureus sortase mutants defective in the display of surface proteins and in the pathogenesis of animal infections.
    Proc Natl Acad Sci U S A. 2000 May 9;97(10):5510-5 PMID: 10805806
  57. Pathogenesis of group A streptococcal infections.
    Clin Microbiol Rev. 2000 Jul;13(3):470-511 PMID: 10885988
  58. A role for the fibrinogen-binding regions of streptococcal M proteins in phagocytosis resistance.
    Mol Microbiol. 2000 Sep;37(6):1318-26 PMID: 10998165
  59. SclA, a novel collagen-like surface protein of Streptococcus pyogenes.
    Infect Immun. 2000 Nov;68(11):6370-7 PMID: 11035747
  60. Identification and characterization of the scl gene encoding a group A Streptococcus extracellular protein virulence factor with similarity to human collagen.
    Infect Immun. 2000 Dec;68(12):6542-53 PMID: 11083763
  61. Inactivation of the srtA gene in Streptococcus gordonii inhibits cell wall anchoring of surface proteins and decreases in vitro and in vivo adhesion.
    Infect Immun. 2001 Jan;69(1):75-80 PMID: 11119491
  62. Streptococcus pyogenes sclB encodes a putative hypervariable surface protein with a collagen-like repetitive structure.
    Microbiology. 2001 Feb;147(Pt 2):419-29 PMID: 11158359
  63. Bacterial determinants of persistent throat colonization and the associated immune response in a primate model of human group A streptococcal pharyngeal infection.
    Cell Microbiol. 2000 Aug;2(4):283-92 PMID: 11207585
  64. Identification and characterization of a second extracellular collagen-like protein made by group A Streptococcus: control of production at the level of translation.
    Infect Immun. 2001 Mar;69(3):1729-38 PMID: 11179350
  65. An embarrassment of sortases - a richness of substrates?
    Trends Microbiol. 2001 Mar;9(3):97-102 PMID: 11239768
  66. Prevention and elimination of upper respiratory colonization of mice by group A streptococci by using a bacteriophage lytic enzyme.
    Proc Natl Acad Sci U S A. 2001 Mar 27;98(7):4107-12 PMID: 11259652
  67. Spa contributes to the virulence of type 18 group A streptococci.
    Infect Immun. 2001 May;69(5):2943-9 PMID: 11292710
  68. Complete genome sequence of an M1 strain of Streptococcus pyogenes.
    Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4658-63 PMID: 11296296
  69. Improved pattern for genome-based screening identifies novel cell wall-attached proteins in gram-positive bacteria.
    Infect Immun. 2001 Jun;69(6):4019-26 PMID: 11349071
  70. Structure of sortase, the transpeptidase that anchors proteins to the cell wall of Staphylococcus aureus.
    Proc Natl Acad Sci U S A. 2001 May 22;98(11):6056-61 PMID: 11371637
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.184.8.2181-2191.2002
Published
2002-04-00
Pages
2181-91
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC134975
Subset
IM
Grants
NIAID NIH HHS · R37 AI020723 · United States
NIAID NIH HHS · R37-AI20723 · United States
External Links
PubMed source DOI: 10.1128/JB.184.8.2181-2191.2002 PMC Full Text View journal details
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