Members of the Streptococcus bovis group are important causes of endocarditis. However, factors associated with their pathogenicity, such as adhesins, remain uncharacterized. We recently demonstrated that endocarditis-derived Streptococcus gallolyticus subsp. gallolyticus isolates frequently adhere to extracellular matrix (ECM) proteins. Here, we generated a draft genome sequence of an ECM protein-adherent S. gallolyticus subsp. gallolyticus strain and found, by genome-wide analyses, 11 predicted LPXTG-type cell wall-anchored proteins with characteristics of MSCRAMMs, including a modular architecture of domains predicted to adopt immunoglobulin (Ig)-like folding. A recombinant segment of one of these, Acb, showed high-affinity binding to immobilized collagen, and cell surface expression of Acb correlated with the presence of acb and collagen adherence of isolates. Three of the 11 proteins have similarities to major pilus subunits and are organized in separate clusters, each including a second Ig-fold-containing MSCRAMM and a class C sortase, suggesting that the sequenced strain encodes three distinct types of pili. Reverse transcription-PCR demonstrated that all three genes of one cluster, acb-sbs7-srtC1, are cotranscribed, consistent with pilus operons of other grampositive bacteria. Further analysis detected expression of all 11 genes in cells grown to mid to late exponential growth phases. Wide distribution of 9 of the 11 genes was observed among S. gallolyticus subsp. gallolyticus isolates with fewer genes present in other S. bovis group species/subspecies. The high prevalence of genes encoding putative MSCRAMMs and pili, including a collagen-binding MSCRAMM, among S. gallolyticus subsp. gallolyticus isolates may play an important role in the predominance of this subspecies in S. bovis endocarditis.Members of the Streptococcus bovis group have long been regarded as opportunistic pathogens and are important causes of endocarditis in humans. These organisms are the most frequently encountered clinical isolates among group D streptococci and the second-most important streptococcal cause of endocarditis after "oral streptococci" (4, 15, 55) which, however, are now classified into many different species and even different genera (16, 28). S. bovis group isolates are considered to be commensals in the intestinal tract, but they have been found only in ϳ2.5% to 15% of the human population (9,24,38,58). Surprisingly, there is a clear correlation-for unknown reasons-between the isolation of S. bovis group organisms as the causative agent of endocarditis and the presence of cancerous lesions and polyps in the intestinal tracts of patients (3,20,24,57), with the first suggestion of this association dating back to 1951 (36). Among patients with carcinoma of the colon, an increased S. bovis carriage rate of 56% was found (24). Whether this increased occurrence is a cause or effect of colon cancer is unclear. However, the association of S. bovis bacteremia or endocarditis with colon cancer raises the intriguing hypothesis...