Consequences of a sortase A mutation in Streptococcus gordonii

Angela H. Nobbs, Reka M. Vajna, Jeremy R. Johnson, Yongshu Zhang, Stanley L. Erlandsen, Monika W. Oli, Jens Kreth, L. Jeannine Brady, Mark C Herzberg

Research output: Contribution to journalArticle

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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 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.

Original languageEnglish (US)
Pages (from-to)4088-4097
Number of pages10
JournalMicrobiology
Volume153
Issue number12
DOIs
StatePublished - Dec 1 2007

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Streptococcus gordonii
Cell Wall
Mutation
Agglutinins
Polystyrenes
Gene Expression Regulation
Biofilms
Immunoblotting
Genes
Polyacrylamide Gel Electrophoresis
Membrane Proteins
Enzyme-Linked Immunosorbent Assay
Antigens
sortase A
Proteins
Bacteria SspA protein

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Nobbs, A. H., Vajna, R. M., Johnson, J. R., Zhang, Y., Erlandsen, S. L., Oli, M. W., ... Herzberg, M. C. (2007). Consequences of a sortase A mutation in Streptococcus gordonii. Microbiology, 153(12), 4088-4097. https://doi.org/10.1099/mic.0.2007/007252-0

Consequences of a sortase A mutation in Streptococcus gordonii. / Nobbs, Angela H.; Vajna, Reka M.; Johnson, Jeremy R.; Zhang, Yongshu; Erlandsen, Stanley L.; Oli, Monika W.; Kreth, Jens; Brady, L. Jeannine; Herzberg, Mark C.

In: Microbiology, Vol. 153, No. 12, 01.12.2007, p. 4088-4097.

Research output: Contribution to journalArticle

Nobbs, AH, Vajna, RM, Johnson, JR, Zhang, Y, Erlandsen, SL, Oli, MW, Kreth, J, Brady, LJ & Herzberg, MC 2007, 'Consequences of a sortase A mutation in Streptococcus gordonii', Microbiology, vol. 153, no. 12, pp. 4088-4097. https://doi.org/10.1099/mic.0.2007/007252-0
Nobbs AH, Vajna RM, Johnson JR, Zhang Y, Erlandsen SL, Oli MW et al. Consequences of a sortase A mutation in Streptococcus gordonii. Microbiology. 2007 Dec 1;153(12):4088-4097. https://doi.org/10.1099/mic.0.2007/007252-0
Nobbs, Angela H. ; Vajna, Reka M. ; Johnson, Jeremy R. ; Zhang, Yongshu ; Erlandsen, Stanley L. ; Oli, Monika W. ; Kreth, Jens ; Brady, L. Jeannine ; Herzberg, Mark C. / Consequences of a sortase A mutation in Streptococcus gordonii. In: Microbiology. 2007 ; Vol. 153, No. 12. pp. 4088-4097.
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