title: Functional analysis of the Streptococcus gordonii DL1 sialic acid-binding adhesin and its essential role in bacterial binding to platelets.
abstract: Bacterial recognition of host sialic acid-containing receptors plays an important role in microbial colonization of the human oral cavity. The sialic acid-binding adhesin of Streptococcus gordonii DL1 was previously associated with the hsa gene encoding a 203-kDa protein. The predicted protein sequence consists of an N-terminal nonrepetitive region (NR1), including a signal sequence, a relatively short serine-rich region (SR1), a second nonrepetitive region (NR2), a long serine-rich region (SR2) containing 113 dodecapeptide repeats, and a C-terminal cell wall anchoring domain. In the present study, the contributions of SR1, NR2, and SR2 to Hsa-mediated adhesion were assessed by genetic complementation. Adhesion of an hsa chromosomal deletion mutant to sialic acid-containing receptors was restored by plasmids containing hsa constructs encoding Hsa that lacked either the N- or C-terminal portion of SR2. In contrast, hsa constructs that lacked the coding sequences for SR1, NR2, or the entire SR2 region failed to restore adhesion. Surface expression of recombinant Hsa was not affected by removal of SR1, NR2, or a portion of SR2 but was greatly reduced by complete removal of SR2. Wheat germ agglutinin, a probe for Hsa-specific glycosylation, reacted with recombinant Hsa lacking SR1, NR2, or SR2 but not with recombinant Hsa lacking both SR1 and SR2. Significantly, the aggregation of human platelets by S. gordonii DL1, an interaction implicated in the pathogenesis of infective endocarditis, required the expression of hsa. Moreover, neuraminidase treatment of the platelets eliminated this interaction, further supporting the hypothesis that Hsa plays an essential role in the bacterium-platelet interaction.
authors.affiliation: Department of Microbiology, The Nippon Dental University School of Dentistry at Tokyo, 1-9-20 Fujimi, Chiyoda-ku, Tokyo 102-8159, Japan. biseibut@tky.ndu.ac.jp
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Ayako
authors.initials: A
authors.lastname: Yajima
authors.forename: John O
authors.initials: JO
authors.lastname: Cisar
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Konishi
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000502:physiology*; D001422:Bacterial Adhesion / Q000502:physiology*; D001792:Blood Platelets / Q000502:physiology*; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology*; D005816:Genetic Complementation Test; D006031:Glycosylation; D006389:Hemagglutinins, Viral; D019158:N-Acetylneuraminic Acid / Q000378:metabolism; D017384:Sequence Deletion; D013291:Streptococcus / Q000502:physiology*; D014909:Wheat Germ Agglutinins / Q000378:metabolism
chemical_list: D018829:Adhesins, Bacterial; D002352:Carrier Proteins; D006389:Hemagglutinins, Viral; D014909:Wheat Germ Agglutinins; C450840:hsa protein, Streptococcus gordonii; D019158:N-Acetylneuraminic Acid