A PTS EII mutant library in Group A Streptococcus identifies a promiscuous man-family PTS transporter influencing SLS-mediated hemolysis

Mol Microbiol. 2017 Feb;103(3):518-533. doi: 10.1111/mmi.13573. Epub 2016 Nov 29.

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bacterial Proteins
  • Carbohydrate Metabolism / genetics
  • Carbohydrate Metabolism / physiology
  • Female
  • Gene Expression Regulation, Bacterial / genetics
  • Gene Library
  • Glucose / metabolism
  • Hemolysis
  • Mannose / metabolism
  • Membrane Transport Proteins / metabolism
  • Mice
  • Operon / genetics
  • Phosphoenolpyruvate Sugar Phosphotransferase System / genetics*
  • Phosphoenolpyruvate Sugar Phosphotransferase System / metabolism
  • Streptococcus / genetics
  • Streptococcus pyogenes / genetics
  • Streptolysins
  • Virulence

Substances

  • Bacterial Proteins
  • Membrane Transport Proteins
  • Streptolysins
  • streptolysin S
  • Phosphoenolpyruvate Sugar Phosphotransferase System
  • phosphoenolpyruvate-mannose phosphotransferase
  • Glucose
  • Mannose