title: Two gene determinants are differentially involved in the biogenesis of Fap1 precursors in Streptococcus parasanguis.
abstract: Mature Fap1, a 200-kDa fimbria-associated adhesin, is required for fimbrial biogenesis and biofilm formation in Streptococcus parasanguis. Fap1-like proteins are found in the genomes of many streptococcal and staphylococcal species. Fap1 is a serine-rich glycoprotein modified by O-linked glycan moieties. In this study, we identified a seven-gene cluster including secY2, orf1, orf2, orf3, secA2, gtf1, and gtf2 that is localized immediately downstream of fap1. The lower G+C contents and the presence of a putative transposase element suggest that this gene cluster was horizontally transferred from other bacteria and represents a genomic island. At least two genes in this island mediated Fap1 biogenesis. Mutation of a glucosyltransferase (Gtf1) gene led to accumulation of a Fap1 precursor, which had no detectable glycan moieties. Inactivation of a gene coding for an accessory Sec protein (SecY2) resulted in expression of a distinct Fap1 precursor, which reacted with one glycan-specific Fap1 antibody but not with another glycan-specific antibody. Furthermore, partially glycosylated Fap1 was detected on the cell surface and in the culture supernatant. These data suggest that SecY2 has a role in complete glycosylation of Fap1 and imply that SecY2 is not the only translocation channel for the Fap1 precursor and that alternative secretion machinery exists. Together, Gtf1 and SecY2 are involved in biogenesis of two distinct Fap1 precursors in S. parasanguis. Discovery of the effect of an accessory Sec protein on Fap1 glycosylation suggests that Fap1 secretion and glycosylation are coupled during Fap1 biogenesis.
authors.affiliation: Department of Pediatric Dentistry, Schools of Dentistry, University of Alabama at Birmingham, Birmingham, AL 35294, USA. hwu@uab.edu
authors.forename: Hui
authors.initials: H
authors.lastname: Wu
authors.forename: Su
authors.initials: S
authors.lastname: Bu
authors.forename: Peter
authors.initials: P
authors.lastname: Newell
authors.forename: Qiang
authors.initials: Q
authors.lastname: Chen
authors.forename: Paula
authors.initials: P
authors.lastname: Fives-Taylor
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D036781:Fimbriae Proteins / Q000096:biosynthesis*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial / Q000235:genetics*; D044404:Genomic Islands; D006031:Glycosylation; D005810:Multigene Family / Q000235:genetics; D013291:Streptococcus / Q000235:genetics* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; C403708:fap1 protein, Streptococcus; C067118:GTF-I protein, Streptococcus; D036781:Fimbriae Proteins