Effect of mutations in Shigella flexneri chromosomal and plasmid-encoded lipopolysaccharide genes on invasion and serum resistance.
This study shows that both length and distribution of lipopolysaccharide (LPS) are important for Shigella flexneri invasion and virulence. Mutants were generated in the chromosomal LPS synthesis genes rfa, rfb, and rol, and in a plasmid-encoded O-antigen chain-length regulator, cld(pHS-2). LPS analysis showed that mutations in rfb genes and in a candidate rfaL gene either eliminated the entire O-antigen side chains or produced chains of greatly reduced length. Mutation in a previously unidentified gene, rfaX, affected the LPS core region and resulted in reduced amounts of O-antigen. Mutants defective in cld(pHS-2) or rol had different distributions of O-antigen chain lengths. The results of tissue-culture cell invasion and plaque assays, the Serény test, and serum-sensitivity assay suggested roles for the different LPS synthesis genes in bacterial survival and virulence; rfaL, rfaX and rfb loci are required for serum resistance and intercellular spread, but not for invasion; cld(pHS-2) is required for resistance to serum killing and for full inflammation in the Serény test, but not for invasion or intercellular spread, while rol is required for normal invasiveness and plaque formation, but not for serum resistance. Thus, O-antigen synthesis and chain-length regulation genes encoded on both the chromosome and the small plasmid pHS-2 play important roles in S. flexneri invasion and virulence.
Hong
M
M
Department of Microbiology, University of Texas at Austin, 78712, USA.
Payne
S M
SM
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D002240:Carbohydrate Sequence; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D006367:HeLa Cells; D006801:Humans; D008025:Ligases / Q000235:genetics; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D051379:Mice; D016254:Mutagenesis, Insertional; D009154:Mutation; D009876:Operon; D010957:Plasmids; D012763:Shigella flexneri / Q000235:genetics* / Q000472:pathogenicity
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D008070:Lipopolysaccharides; D008565:Membrane Proteins; C103853:RfaL protein, E coli; C489586:wzzB protein, E coli; C075969:rol protein, Bacteria; D008025:Ligases