TY - JOUR
T1 - Consequences of a sortase A mutation in Streptococcus gordonii
AU - Nobbs, Angela H.
AU - Vajna, Reka M.
AU - Johnson, Jeremy R.
AU - Zhang, Yongshu
AU - Erlandsen, Stanley L.
AU - Oli, Monika W.
AU - Kreth, Jens
AU - Brady, L. Jeannine
AU - Herzberg, Mark C.
PY - 2007/12
Y1 - 2007/12
N2 - Sortase A (SrtA) is required for cell-wall anchoring of LPXTG-containing Gram-positive surface proteins. It was hypothesized, therefore, that disruption of the srtA gene would alter surface anchoring and functions of target LPXTG motif-bearing SspA and SspB proteins of Streptococcus gordonii. Mutant strains in srtA (V288srtA-, DL1 srtA-) were constructed in S. gordanii V288 (wtV288) and DL1 (wtDL1). When compared to wtV288, the V288srtA-mutant showed decreased biofilm formation on polystyrene, and reduced binding to immobilized purified salivary agglutinin (BIAcore analysis). The wtV288 and V288srtA-strains were similar in ultrastructure, but immunogold-labelled SspA/SspB surface expression was reduced on the V288srtA-mutant. DL1srtA- was also complemented to obtain DL1srtA+. From the wild-type strains (wtV288, wtDL1), srtA-mutants (V288srtA-, DL1 srtA-), and the complemented mutant (DL1srtA+), cytoplasmic, cell-wall and released extracellular protein fractions were isolated. Each fraction was analysed by SDS-PAGE and immunoblotting with anti-P1. Spent medium from srtA-mutant cells contained over-represented proteins, including SspA/SspB (P1 antigen). Mutants showed less P1 on the cell surface than wild-types, as estimated using whole-cell ELISA, and no P1 appeared in the cytoplasmic fractions. Expression of several adhesin genes (sspA/B, cshA/B, fbpA) was generally upregulated in the mutants (V288srtA-, DL1srtA-), but restored to wild-type levels in DL1srtA+. These data therefore imply that in addition to its role in processing LPXTG-containing adhesins, sortase A has the novel function of contributing to transcriptional regulation of adhesin gene expression.
AB - Sortase A (SrtA) is required for cell-wall anchoring of LPXTG-containing Gram-positive surface proteins. It was hypothesized, therefore, that disruption of the srtA gene would alter surface anchoring and functions of target LPXTG motif-bearing SspA and SspB proteins of Streptococcus gordonii. Mutant strains in srtA (V288srtA-, DL1 srtA-) were constructed in S. gordanii V288 (wtV288) and DL1 (wtDL1). When compared to wtV288, the V288srtA-mutant showed decreased biofilm formation on polystyrene, and reduced binding to immobilized purified salivary agglutinin (BIAcore analysis). The wtV288 and V288srtA-strains were similar in ultrastructure, but immunogold-labelled SspA/SspB surface expression was reduced on the V288srtA-mutant. DL1srtA- was also complemented to obtain DL1srtA+. From the wild-type strains (wtV288, wtDL1), srtA-mutants (V288srtA-, DL1 srtA-), and the complemented mutant (DL1srtA+), cytoplasmic, cell-wall and released extracellular protein fractions were isolated. Each fraction was analysed by SDS-PAGE and immunoblotting with anti-P1. Spent medium from srtA-mutant cells contained over-represented proteins, including SspA/SspB (P1 antigen). Mutants showed less P1 on the cell surface than wild-types, as estimated using whole-cell ELISA, and no P1 appeared in the cytoplasmic fractions. Expression of several adhesin genes (sspA/B, cshA/B, fbpA) was generally upregulated in the mutants (V288srtA-, DL1srtA-), but restored to wild-type levels in DL1srtA+. These data therefore imply that in addition to its role in processing LPXTG-containing adhesins, sortase A has the novel function of contributing to transcriptional regulation of adhesin gene expression.
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U2 - 10.1099/mic.0.2007/007252-0
DO - 10.1099/mic.0.2007/007252-0
M3 - Article
C2 - 18048922
AN - SCOPUS:37449026635
SN - 1350-0872
VL - 153
SP - 4088
EP - 4097
JO - Microbiology
JF - Microbiology
IS - 12
ER -