Mouse skin passage of a Streptococcus pyogenes Tn917 mutant of sagA/pel restores virulence, beta-hemolysis and sagA/pel expression without altering the position or sequence of the transposon

BMC Microbiol. 2001:1:33. doi: 10.1186/1471-2180-1-33. Epub 2001 Dec 17.

Abstract

Background: Streptolysin S (SLS), the oxygen-stable hemolysin of Streptococcus pyogenes, has recently been shown to be encoded by the sagA/pel gene. Mutants lacking expression of this gene were less virulent in a dermonecrotic mouse infection model. Inactivation of the sagA/pel gene affect the expression of a variety of virulence factors in addition to the hemolysin. Insertion of a Tn917 transposon into the promoter region of the sagA/pel gene of S. pyogenes isolate CS101 eliminated expression of SLS, as well as decreased expression of the streptococcal pyrogenic exotoxin B, streptokinase and M protein.

Results: In this study a mouse skin air sac model was utilized to analyze the effect of biological pressures on expression of SLS and other sagA/pel regulated gene products. The insertion delayed the lethal effect of S. pyogenes in a mouse skin infection model. Despite this, bacteria could be cultured from the kidneys 72 hours post infection. These kidney-recovered isolates were beta-hemolytic despite the transposon being present in its original location and had equivalent virulence to the wild type isolate when re-injected into naive mice. Northern blot analysis of the kidney-recovered isolates confirmed that transcription of sagA/pel was restored; however the expression of all sagA/pel regulated genes was not restored to wild type levels.

Conclusions: These results show that biological pressure present in the mouse can select for variants with altered expression of key virulence factor genes in S. pyogenes.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • DNA Transposable Elements / genetics*
  • Fungal Proteins / biosynthesis*
  • Fungal Proteins / genetics
  • Gene Expression
  • Hemolysis
  • Mice
  • Mutation
  • Polysaccharide-Lyases / biosynthesis*
  • Polysaccharide-Lyases / genetics
  • Serial Passage / methods
  • Skin / microbiology
  • Streptococcus pyogenes / genetics
  • Streptococcus pyogenes / pathogenicity*
  • Virulence / genetics*

Substances

  • DNA Transposable Elements
  • Fungal Proteins
  • SAGA protein, Emericella nidulans
  • Polysaccharide-Lyases
  • pectate lyase