Genetic loci of Streptococcus mitis that mediate binding to human platelets.
The direct binding of bacteria to platelets is a postulated major interaction in the pathogenesis of infective endocarditis. To identify bacterial components that mediate platelet binding by Streptococcus mitis, we screened a Tn916deltaE-derived mutant library of S. mitis strain SF100 for reduced binding to human platelets in vitro. Two distinct loci were found to affect platelet binding. The first contains a gene (pblT) encoding a highly hydrophobic, 43-kDa protein with 12 potential membrane-spanning segments. This protein resembles members of the major facilitator superfamily of small-molecule transporters. The second platelet binding locus consists of an apparent polycistronic operon. This region includes genes that are highly similar to those of Lactococcus lactis phage r1t and Streptococcus thermophilus phage 01205. Two genes (pblA and pblB) encoding large surface proteins are also present. The former encodes a 107-kDa protein containing tryptophan-rich repeats, which may serve to anchor the protein within the cell wall. The latter encodes a 121-kDa protein most similar to a tail fiber protein from phage 01205. Functional mapping by insertion-duplication mutagenesis and gene complementation indicates that PblB may be a platelet adhesin and that expression of PblB may be linked to that of PblA. The combined data indicate that at least two genomic regions contribute to platelet binding by S. mitis. One encodes a probable transmembrane transporter, while the second encodes two large surface proteins resembling structural components of lysogenic phages.
Bensing
B A
BA
Veterans Affairs Medical Center and University of California San Francisco, San Francisco, California 94121, USA.
Rubens
C E
CE
Sullam
P M
PM
D000595:Amino Acid Sequence; D001422:Bacterial Adhesion / Q000235:genetics*; D001483:Base Sequence; D001792:Blood Platelets / Q000382:microbiology*; D002352:Carrier Proteins / Q000235:genetics; D004697:Endocarditis, Bacterial / Q000382:microbiology; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005814:Genes, Viral; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013291:Streptococcus / Q000235:genetics*; D017108:Streptococcus Phages / Q000235:genetics; D015678:Viral Structural Proteins / Q000235:genetics
D002352:Carrier Proteins; D008565:Membrane Proteins; D015678:Viral Structural Proteins