Molecular and chemical characterization of the lipopolysaccharide O-antigen and its role in the virulence of Yersinia enterocolitica serotype O:8

Mol Microbiol. 1997 Jan;23(1):63-76. doi: 10.1046/j.1365-2958.1997.1871558.x.

Abstract

The Y. enterocolitica O:8(YeO8) O-antigen repeat units consist of five sugar residues: N-acetyl-D-galactosamine (GalNAc), D-galactose (Gal), D-mannose (Man), L-fucose (Fuc), and 6-deoxy-D-gulose (6d-Gul). The nucleotide sequence of the O-antigen gene cluster of the YeO8 strain 8081-c was determined. Altogether, 18 open reading frames (ORFs) were identified and shown to be essential for O-antigen biosynthesis. We previously characterized the 3'-end of the O-antigen gene cluster and identified four genes: two for GDP-Man biosynthesis, one for UDP-Gal biosynthesis, and one for O-antigen polymerase. Based on sequence similarity, Tn5-insertion phenotypes and chemical analysis, the 14 new genes were assigned the following functions: four genes are involved in the biosynthesis of CDP-6d-Gul and two in GDP-Fuc biosynthesis. Five gene products were assigned sugar transferase functions and one gene product was similar to Wzx, the O-antigen flippase. Two genes remained unassigned. By genetic complementation we also showed that YeO8 O-antigen biosynthesis was dependent on N-acetyl-glucosaminyl:undecaprenylphosphate transferase (GlcNAc transferase), the WecA (formerly known as Rfe) protein. Data obtained from chemical-composition analysis suggest that in addition to being GlcNAc transferase, WecA may also function as a GalNAc transferase. Using a restriction-deficient derivative of Y. enterocolitica O:8 strain 8081, a rough mutant, designated 8081-R2, was isolated. 8081-R2 was complemented in trans with a cloned O-antigen gene cluster restoring surface O-antigen expression. The virulence of the wild-type strain and that of the complemented strain were significantly higher (approx. 100-fold) than that of the rough mutant in an orally infected mouse model, showing that YeO8 O-antigen is a virulence factor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacteriophages
  • Base Sequence
  • DNA Transposable Elements
  • DNA, Bacterial
  • Escherichia coli / metabolism
  • Genetic Complementation Test
  • Lipopolysaccharides / chemistry
  • Mice
  • Mice, Inbred DBA
  • Molecular Sequence Data
  • Multigene Family*
  • Mutagenesis, Insertional
  • N-Acetylglucosaminyltransferases / metabolism
  • O Antigens*
  • Phenotype
  • Receptors, Virus
  • Serotyping
  • Transferases / genetics
  • Virulence
  • Yersinia enterocolitica / pathogenicity*

Substances

  • DNA Transposable Elements
  • DNA, Bacterial
  • Lipopolysaccharides
  • O Antigens
  • Receptors, Virus
  • Transferases
  • N-Acetylglucosaminyltransferases
  • UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase

Associated data

  • GENBANK/B42476
  • GENBANK/D21242
  • GENBANK/L01777
  • GENBANK/L41518
  • GENBANK/P09147
  • GENBANK/P19526
  • GENBANK/P22716
  • GENBANK/P26394
  • GENBANK/P26396
  • GENBANK/P26397
  • GENBANK/P26401
  • GENBANK/P29783
  • GENBANK/P32054
  • GENBANK/P32055
  • GENBANK/P33699
  • GENBANK/P37780
  • GENBANK/P37781
  • GENBANK/Q03583
  • GENBANK/S22617
  • GENBANK/S27581
  • GENBANK/S46493
  • GENBANK/S73325
  • GENBANK/U24571
  • GENBANK/U25839
  • GENBANK/U32832
  • GENBANK/U46859
  • GENBANK/X77617
  • GENBANK/X80225
  • GENBANK/Z47767