A plasmid-borne O-antigen chain length determinant and its relationship to other chain length determinants

Stevenson, Gordon; Kessler, Annette C.; Reeves, Peter R.

doi:10.1111/j.1574-6968.1995.tb07330.x

Cited by 53 publications

(49 citation statements)

References 23 publications

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“…The later finding of its homology to cld/rol genes, a family of O-antigen chain-length determinants (Stevenson et al, 1995), led us to include this gene in our study of Shigella LPS. Several strains were constructed in order to study the expression and localization of the gene product, and the role of cld pHS-2 in virulence.…”

Section: Isolation Of Strains With Chromosomal Lps-synthesis Gene Mutmentioning

confidence: 99%

“…In both cases, loss of the plasmid-encoded O-antigen genes resulted in loss of virulence (Kopecko et al, 1980;. S. flexneri serotype 2a isolates often contain a 3 kb plasmid, pHS-2 (Stieglitz et al, 1989), which has an open reading frame (ORF) for a predicted 368-amino-acid protein (Stieglitz et al, 1989;Stevenson et al, 1995). The predicted protein has 63% amino acid identity with E. coli FepE (C. Shea-Cleavinger, B. Ozenberger and M. McIntosh, Genbank Accession No.…”

Section: Introductionmentioning

confidence: 99%

“…Although fepE is located within the enterobactin biosynthesis and transport cluster on the E. coli chromosome (Ozenberger et al, 1987), it does not play an essential role in this iron-transport system in E. coli (M. McIntosh, personal communication). Recently it was shown that the predicted protein sequences of both the pHS-2-encoded gene and fepE have 21-26% identity to those of a family of chromosomal genes determining LPS O-antigen chain length (Batchelor et al, 1992;Bastin et al, 1993;Morona et al, 1995;Stevenson et al, 1995). This family has been called cld (O antigen chain-length determinant), rol (regulation of O-antigen chain length), or wzz in different E. coli, Salmonella, and Shigella species (Reeves et al, 1996;Whitfield et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

“…This family has been called cld (O antigen chain-length determinant), rol (regulation of O-antigen chain length), or wzz in different E. coli, Salmonella, and Shigella species (Reeves et al, 1996;Whitfield et al, 1997). The plasmid pHS-2-encoded gene was named cld pHS-2 (Stevenson et al, 1995). It was shown that the absence of this gene caused loss of O-antigen chains of 90-100 repeating units in S. flexneri serotype 2a (Stevenson et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

“…The plasmid pHS-2-encoded gene was named cld pHS-2 (Stevenson et al, 1995). It was shown that the absence of this gene caused loss of O-antigen chains of 90-100 repeating units in S. flexneri serotype 2a (Stevenson et al, 1995). It was not reported if this plasmid-encoded gene plays a role in S. flexneri virulence.…”

Section: Introductionmentioning

confidence: 99%

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Effect of mutations in Shigella flexneri chromosomal and plasmid‐encoded lipopolysaccharide genes on invasion and serum resistance

Hong

Payne

1997

Molecular Microbiology

117

120

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SummaryThis 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 Seré ny 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 Seré ny 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.

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Section: Isolation Of Strains With Chromosomal Lps-synthesis Gene Mutmentioning

confidence: 99%