title: A PTS EII mutant library in Group A Streptococcus identifies a promiscuous man-family PTS transporter influencing SLS-mediated hemolysis.
abstract: The Group A Streptococcus (GAS, Streptococcus pyogenes) is a Gram-positive human pathogen that must adapt to unique host environments in order to survive. Links between sugar metabolism and virulence have been demonstrated in GAS, where mutants in the phosphoenolpyruvate-dependent phosphotransferase system (PTS) exhibited Streptolysin S (SLS)-mediated hemolysis during exponential growth. This early onset hemolysis correlated with an increased lesion size and severity in a murine soft tissue infection model when compared with parental M1T1 MGAS5005. To identify the PTS components responsible for this phenotype, we insertionally inactivated the 14 annotated PTS EIIC-encoding genes in the GAS MGAS5005 genome and subjected this library to metabolic and hemolysis assays to functionally characterize each EIIC. It was found that a few EIIs had a very limited influence on PTS sugar metabolism, whereas others were fairly promiscuous. The mannose-specific EII locus, encoded by manLMN, was expressed as a mannose-inducible operon that exhibited the most influence on PTS sugar metabolism, including mannose. Importantly, components of the mannose-specific EII also acted to prevent the early onset of SLS-mediated hemolysis. Interestingly, these roles were not identical in two different M1T1 GAS strains, highlighting the possible versatility of the PTS to adapt to strain-specific needs.
authors.affiliation: Department of Cell Biology and Molecular Genetics, Maryland Pathogen Research Institute, University of Maryland, College Park, MD, 20742, USA.
authors.forename: Ganesh S
authors.initials: GS
authors.lastname: Sundar
authors.affiliation: Department of Cell Biology and Molecular Genetics, Maryland Pathogen Research Institute, University of Maryland, College Park, MD, 20742, USA.
authors.forename: Emrul
authors.initials: E
authors.lastname: Islam
authors.affiliation: Department of Cell Biology and Molecular Genetics, Maryland Pathogen Research Institute, University of Maryland, College Park, MD, 20742, USA.
authors.forename: Kanika
authors.initials: K
authors.lastname: Gera
authors.affiliation: Department of Cell Biology and Molecular Genetics, Maryland Pathogen Research Institute, University of Maryland, College Park, MD, 20742, USA.
authors.forename: Yoann
authors.initials: Y
authors.lastname: Le Breton
authors.affiliation: Department of Cell Biology and Molecular Genetics, Maryland Pathogen Research Institute, University of Maryland, College Park, MD, 20742, USA.
authors.forename: Kevin S
authors.initials: KS
authors.lastname: McIver
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D050260:Carbohydrate Metabolism / Q000235:genetics / Q000502:physiology; D005260:Female; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D015723:Gene Library; D005947:Glucose / Q000378:metabolism; D006461:Hemolysis; D008358:Mannose / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000378:metabolism; D051379:Mice; D009876:Operon / Q000235:genetics; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics* / Q000378:metabolism; D013291:Streptococcus / Q000235:genetics; D013297:Streptococcus pyogenes / Q000235:genetics; D013301:Streptolysins; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D013301:Streptolysins; C026050:streptolysin S; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; C037992:phosphoenolpyruvate-mannose phosphotransferase; D005947:Glucose; D008358:Mannose