Distribution of the rol gene encoding the regulator of lipopolysaccharide O-chain length in Escherichia coli and its influence on the expression of group I capsular K antigens
- 1 April 1996
- journal article
- Published by American Society for Microbiology in Journal of Bacteriology
- Vol. 178 (7) , 1895-1902
- https://doi.org/10.1128/jb.178.7.1895-1902.1996
Abstract
The rol (cld) gene encodes a protein involved in the expression of lipopolysaccharides in some members of the family Enterobacteriaceae. Rol interacts with one or more components of Rfc-dependent O-antigen biosynthetic complexes to regulate the chain length of lipopolysaccharide O antigens. The Rfc-Rol-dependent pathway for O-antigen synthesis is found in strains with heteropolysaccharide O antigens, and, consistent with this association, rol-homologous sequences were detected in chromosomal DNAs from 17 different serotypes with heteropolysaccharide O antigens. Homopolymer O antigens are synthesized by a pathway that does not involve either Rfc or Rol. It was therefore unexpected when a survey of Escherichia coli strains possessing mannose homopolymer O8 and O9 antigens showed that some strains contained rol. All 11 rol-positive strains coexpressed a group IB capsular K antigen with the O8 or O9 antigen. In contrast, 12 rol-negative strains all produced group IA K antigens in addition to the homopolymer O antigen. Previous research from this and other laboratories has shown that portions of the group I K antigens are attached to lipopolysaccharide lipid A-core, in a form that we have designated K(LPS). By constructing a hybrid strain with a deep rough rfa defect, it was shown that the K40 (group IB) K(LPS) antigen exists primarily as long chains. However, a significant amount of K40 antigen was surface expressed in a lipid A-core-independent pathway. The typical chain length distribution of the K40 antigen was altered by introduction of multicopy rol, suggesting that the K40 group IB K antigen is equivalent to a Rol-dependent O antigen. The prototype K30 (group IA) K antigen is expressed as short oligosaccharides (primarily single repeat units) in K(LPS), as well as a high-molecular-weight lipid A-core-independent form. Introduction of multicopy rol into the K30 strain generated a novel modal pattern of K(LPS) with longer polysaccharide chains. Collectively, these results suggested that group IA K(LPS) is also synthesized by a Rol-dependent pathway and that the typically short oligosaccharide K(LPS) results from the absence of Rol activity in these strains.Keywords
This publication has 56 references indexed in Scilit:
- A simple procedure to demonstrate the presence of the 0-antigen capsule in enteropathogenic Escherichia coliZentralblatt für Bakteriologie, 1994
- Function of the rfb gene cluster and the rfe gene in the synthesis of O antigen by Shigella dysenteriae 1Molecular Microbiology, 1993
- Core-lipid A on the K40 Polysaccharide of Escherichia coli O8: K40: H9, a Representative of Group I Capsular PolysaccharidesZentralblatt für Bakteriologie, 1992
- The bex locus in encapsulated Haemophilus influenzae: a chromosomal region involved in capsule polysaccharide exportMolecular Microbiology, 1990
- Use of a bacteriophage-encoded glycanase enzyme in the generation of lipopolysaccharide O side chain deficient mutants of Escherichia coli O9:K30 and Klebsiella O1:K20: role of O and K antigens in resistance to complement-mediated serum killingCanadian Journal of Microbiology, 1989
- Mutants of Escherichia coli O9: K30 with Altered Synthesis and Expression of the Capsular K30 AntigenMicrobiology, 1989
- ECA, the enterobacterial common antigenFEMS Microbiology Letters, 1988
- Transfer and expression of the genetic determinants for O and K antigen synthesis in Escherichia coli O9:K(A)30 and Klebsiella sp. Ol:K20, in Escherichia coli K12Canadian Journal of Microbiology, 1988
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979
- Genetic Determinants of the Synthesis of the Polysaccharide Capsular Antigen K27(A) of Escherichia coliJournal of General Microbiology, 1977