title: 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.
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.
authors.affiliation: Department of Microbiology and Immunology, Medical College of Ohio, Toledo, Ohio 43614, USA. teberhard@mco.edu
authors.forename: T H
authors.initials: TH
authors.lastname: Eberhard
authors.forename: D D
authors.initials: DD
authors.lastname: Sledjeski
authors.forename: M D
authors.initials: MD
authors.lastname: Boyle
mesh_terms: D000818:Animals; D004251:DNA Transposable Elements / Q000235:genetics*; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D006461:Hemolysis; D051379:Mice; D009154:Mutation; D011133:Polysaccharide-Lyases / Q000096:biosynthesis* / Q000235:genetics; D012692:Serial Passage / Q000379:methods; D012867:Skin / Q000382:microbiology; D013297:Streptococcus pyogenes / Q000235:genetics / Q000472:pathogenicity*; D014774:Virulence / Q000235:genetics*
chemical_list: D004251:DNA Transposable Elements; D005656:Fungal Proteins; C118425:SAGA protein, Emericella nidulans; D011133:Polysaccharide-Lyases; C023911:pectate lyase