title: Both GtfA and GtfB are required for SraP glycosylation in Staphylococcus aureus.
abstract: Staphylococcus aureus has been shown to bind to human platelets through a variety of surface molecules, including serine-rich adhesin for platelets (SraP). The SraP mutant strain of S. aureus is significantly impaired in its ability to initiate infection compared with the wild strain. SraP is a cell wall-anchored, glycosylated protein. A previous study revealed that SecY2, Asp1, Asp2, Asp3, and SecA2 in the SraP operon were required for the efficient transport of glycosylated SraP from the cytoplasm to the bacterial cell surface. However, no glycosyltransferase (Gtf) was found to be involved in the glycosylation of SraP. In this study, SraP was found in all of the 55 clinical isolates of S. aureus using a real-time polymerase chain reaction assay. Sequence and phylogenetic analysis showed that GtfA and GtfB in the SraP operon were highly conserved in most of these clinical isolates. Conserved domains analysis revealed that both GtfA and GtfB contained a GT1_GtfA-like domain. Structural homology analysis inferred that they are both Gtfs. We then constructed an in vivo glycosylation system in Escherichia coli using SraP1–743 as the substrate and GtfA and GtfB as the Gtfs. Using this system, we found that GtfA and GtfB were the Gtfs that transferred the N-acetylglucosamine-containing oligosaccharides to the recombinant SraP1–743. Deletion of either one or both of the Gtfs abolished the glycosylation of SraP. In summary, GtfA and GtfB in the SraP operon are highly conserved in most clinical isolates of S. aureus, and both GtfA and GtfB are required for SraP glycosylation.
authors.forename: Yirong
authors.initials: Y
authors.lastname: Li
authors.forename: Xiang
authors.initials: X
authors.lastname: Huang
authors.forename: Jingjing
authors.initials: J
authors.lastname: Li
authors.forename: Ji
authors.initials: J
authors.lastname: Zeng
authors.forename: Fan
authors.initials: F
authors.lastname: Zhu
authors.forename: Wen
authors.initials: W
authors.lastname: Fan
authors.forename: Lihua
authors.initials: L
authors.lastname: Hu
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D017124:Conserved Sequence; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D009876:Operon; D010802:Phylogeny; D011487:Protein Conformation; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D013211:Staphylococcus aureus / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D005964:Glucosyltransferases