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Volume 153, Issue 12

Consequences of a sortase A mutation in Free

Abstract

Sortase A (SrtA) is required for cell-wall anchoring of LPXTG-containing Gram-positive surface proteins. It was hypothesized, therefore, that disruption of the gene would alter surface anchoring and functions of target LPXTG motif-bearing SspA and SspB proteins of . Mutant strains in (V288 , DL1) were constructed in V288 (wtV288) and DL1 (wtDL1). When compared to wtV288, the V288 mutant showed decreased biofilm formation on polystyrene, and reduced binding to immobilized purified salivary agglutinin (BIAcore analysis). The wtV288 and V288 strains were similar in ultrastructure, but immunogold-labelled SspA/SspB surface expression was reduced on the V288 mutant. DL1 was also complemented to obtain DL1. From the wild-type strains (wtV288, wtDL1), mutants (V288, DL1), and the complemented mutant (DL1), 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 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 (/, /, ) was generally upregulated in the mutants (V288, DL1), but restored to wild-type levels in DL1. 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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Keyword(s): DL1srtA+, complemented DL1srtA− , DL1srtA−, sortase A negative mutant in Streptococcus gordonii DL1 wild-type , P1, antigen class I/II surface protein of Streptococcus mutans , RU, resonance units , V288srtA−, sortase A negative mutant in S. gordonii V288 wild-type , wtDL1, S. gordonii DL1 wild-type and wtV288, S. gordonii V288 wild-type
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2007-12-01
2024-04-19
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Consequences of a sortase A mutation in Streptococcus gordonii
Microbiology 153, 4088 (2007); https://doi.org/10.1099/mic.0.2007/007252-0
/content/journal/micro/10.1099/mic.0.2007/007252-0
/content/journal/micro/10.1099/mic.0.2007/007252-0
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