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Streptococcus pyogenes cytolysin-mediated translocation does not require pore formation by streptolysin O.

ABSTRACT: Bacterial toxin injection into the host cell is required for the virulence of numerous pathogenic bacteria. Cytolysin-mediated translocation (CMT) of Streptococcus pyogenes uses streptolysin O (SLO) to translocate the S. pyogenes nicotinamide adenine dinucleotide-glycohydrolase (SPN) into the host cell cytosol, resulting in the death of the host cell. Although SLO is a pore-forming protein, previous studies have shown that pore formation alone is not sufficient for CMT to occur. Thus, the role and requirement of the SLO pore remains unclear. In this study, we constructed various S. pyogenes strains expressing altered forms of SLO to assess the importance of pore formation. We observed that SLO mutants that are unable to form pores retain the ability to translocate SPN. In addition, SPN translocation occurs after inhibition of actin polymerization, suggesting that CMT occurs independently of clathrin-mediated endocytosis. Moreover, despite the ability of mutants to translocate SPN, their cytotoxic effect requires SLO pore formation.

SUBMITTER: Magassa N

PROVIDER: S-EPMC2868537 | biostudies-other | 2010 May

REPOSITORIES: biostudies-other

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Streptococcus pyogenes cytolysin-mediated translocation does not require pore formation by streptolysin O.

Magassa N'Goundo N Chandrasekaran Sukantha S Caparon Michael G MG

EMBO reports 20100326 5

Bacterial toxin injection into the host cell is required for the virulence of numerous pathogenic bacteria. Cytolysin-mediated translocation (CMT) of Streptococcus pyogenes uses streptolysin O (SLO) to translocate the S. pyogenes nicotinamide adenine dinucleotide-glycohydrolase (SPN) into the host cell cytosol, resulting in the death of the host cell. Although SLO is a pore-forming protein, previous studies have shown that pore formation alone is not sufficient for CMT to occur. Thus, the role a ...[more]

PMID: 20339385

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Project description:Biofilms play an important role in the pathogenesis of group A streptococcus (GAS), a Gram-positive pathogen responsible for a wide range of infections and with a significant public health impact. Although most GAS serotypes are able to form biofilms, there is a large amount of heterogeneity between individual strains in biofilm formation, as measured by standard crystal violet assays. It is generally accepted that biofilm formation includes the initial adhesion of bacterial cells to a surface followed by microcolony formation, biofilm maturation, and extensive production of extracellular matrix that links together proliferating cells and provides a scaffold for the three-dimensional (3D) biofilm structure. However, our studies show that for GAS strain JS95, microcolony formation is not an essential step in static biofilm formation, and instead, biofilm can be effectively formed from slow-growing or nonreplicating late-exponential- or early-stationary-phase planktonic cells via sedimentation and fixation of GAS chains. In addition, we show that the GAS capsule specifically contributes to the alternative sedimentation-initiated biofilms. Microcolony-independent sedimentation biofilms are similar in morphology and 3D structure to biofilms initiated by actively dividing planktonic bacteria. We conclude that GAS can form biofilms by an alternate noncanonical mechanism that does not require transition from microcolony formation to biofilm maturation and which may be obscured by biofilm phenotypes that arise via the classical biofilm maturation processes.IMPORTANCE The static biofilm assay is a common tool for easy biomass quantification of biofilm-forming bacteria. However, Streptococcus pyogenes biofilm formation as measured by the static assay is strain dependent and yields heterogeneous results for different strains of the same serotype. In this study, we show that two independent mechanisms, for which the protective capsule contributes opposing functions, may contribute to static biofilm formation. We propose that separation of these mechanisms for biofilm formation might uncover previously unappreciated biofilm phenotypes that may otherwise be masked in the classic static assay.

Dataset Information

Streptococcus pyogenes cytolysin-mediated translocation does not require pore formation by streptolysin O.

Publications

Streptococcus pyogenes cytolysin-mediated translocation does not require pore formation by streptolysin O.

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