Sortase A utilizes an ancillary protein anchor for efficient cell wall anchoring of pili in Streptococcus agalactiae.
Pili are putative virulence factors and promising vaccine candidates in Streptococcus agalactiae (group B Streptococcus [GBS]) infection, a leading cause of neonatal sepsis and meningitis. The genes necessary for pilus synthesis and assembly are clustered in pilus islands (PI). Each gene encodes three structural subunits (a backbone and two ancillary proteins) bearing a C-terminal LPXTG motif and two subfamily C sortases (SrtC) involved in covalent polymerization of the subunits. GBS strains also possess the conserved "housekeeping" sortase A (SrtA), but its role in pilus assembly is unclear. To address this issue, pilus expression and cell wall anchoring were analyzed in srtA deletion mutants. Loss of SrtA did not affect pilus polymerization. However, pilus expression on the cell surface was reduced, and pili accumulated in the culture supernatant. Furthermore, cell-associated pili could be readily released by detergent treatment, indicating that SrtA is involved in covalent anchoring of pili to the cell wall. When each of the genes comprising PI-2a was systematically deleted, only the absence of ancillary subunit GBS150 or the SrtC required for incorporation of GBS150 into pili mimicked the srtA mutant phenotype. Thus, from these data a model for GBS pilus assembly can be proposed in which PI sortases are responsible for polymerization of the pilus structure, while SrtA is required to covalently attach it to the cell wall, utilizing ancillary pilus subunit GBS150 as the anchor protein.
Novartis Vaccines, Via Fiorentina 1, 53100 Siena, Italy.
Angela H
AH
Nobbs
Roberto
R
Rosini
C Daniela
CD
Rinaudo
Domenico
D
Maione
Guido
G
Grandi
John L
JL
Telford
D019881:Aminoacyltransferases / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000378:metabolism*; D017353:Gene Deletion; D023061:Gene Order; D005810:Multigene Family; D013292:Streptococcus agalactiae / Q000235:genetics / Q000502:physiology*
D001426:Bacterial Proteins; C509822:sortase C; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases