Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Oct;178(20):5916–5924. doi: 10.1128/jb.178.20.5916-5924.1996

Identification of the galE gene and a galE homolog and characterization of their roles in the biosynthesis of lipopolysaccharide in a serotype O:8 strain of Yersinia enterocolitica.

D E Pierson 1, S Carlson 1
PMCID: PMC178447  PMID: 8830687

Abstract

A clone that complements mutations in Yersinia enterocolitica lipopolysaccharide (LPS) core biosynthesis was isolated, and the DNA sequence of the clone was determined. Three complete open reading frames and one partial open reading frame were located on the cloned DNA fragment. The first, partial, open reading frame had homology to the rfbK gene. The remaining reading frames had homology to galE, rol, and gsk. Analysis of the galE homolog indicates that although it can complement an Escherichia coli galE mutant, its primary function in Y. enterocolitica is not in the production of UDP galactose but, instead, some other nucleotide sugar required for LPS biosynthesis. This gene has been renamed lse, for LPS sugar epimerase. The rol homolog has been demonstrated to have a role in Y. enterocolitica serotype 0:8 O-polysaccharide antigen chain length determination. An additional galE homolog has been identified in Y. enterocolitica by homology to the E. coli gene. The product of this gene has UDP galactose 4-epimerase activity in both E. coli and Y. enterocolitica. This gene is linked to the other genes of the galactose utilization pathway, similar to what is seen in other members of the family Enterobacteriaceae. Although Y. enterocolitica 0:8 strains are reported to have galactose as a constituent of LPS, a strain containing a mutation in this galE gene does not exhibit any LPS defects.

Full Text

The Full Text of this article is available as a PDF (3.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bastin D. A., Stevenson G., Brown P. K., Haase A., Reeves P. R. Repeat unit polysaccharides of bacteria: a model for polymerization resembling that of ribosomes and fatty acid synthetase, with a novel mechanism for determining chain length. Mol Microbiol. 1993 Mar;7(5):725–734. doi: 10.1111/j.1365-2958.1993.tb01163.x. [DOI] [PubMed] [Google Scholar]
  2. Batchelor R. A., Alifano P., Biffali E., Hull S. I., Hull R. A. Nucleotide sequences of the genes regulating O-polysaccharide antigen chain length (rol) from Escherichia coli and Salmonella typhimurium: protein homology and functional complementation. J Bacteriol. 1992 Aug;174(16):5228–5236. doi: 10.1128/jb.174.16.5228-5236.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Batchelor R. A., Haraguchi G. E., Hull R. A., Hull S. I. Regulation by a novel protein of the bimodal distribution of lipopolysaccharide in the outer membrane of Escherichia coli. J Bacteriol. 1991 Sep;173(18):5699–5704. doi: 10.1128/jb.173.18.5699-5704.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bauer A. J., Rayment I., Frey P. A., Holden H. M. The molecular structure of UDP-galactose 4-epimerase from Escherichia coli determined at 2.5 A resolution. Proteins. 1992 Apr;12(4):372–381. doi: 10.1002/prot.340120409. [DOI] [PubMed] [Google Scholar]
  5. Bliska J. B., Guan K. L., Dixon J. E., Falkow S. Tyrosine phosphate hydrolysis of host proteins by an essential Yersinia virulence determinant. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1187–1191. doi: 10.1073/pnas.88.4.1187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bouffard G. G., Rudd K. E., Adhya S. L. Dependence of lactose metabolism upon mutarotase encoded in the gal operon in Escherichia coli. J Mol Biol. 1994 Dec 2;244(3):269–278. doi: 10.1006/jmbi.1994.1728. [DOI] [PubMed] [Google Scholar]
  7. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haber R., Adhya S. Interaction of spatially separated protein-DNA complexes for control of gene expression: operator conversions. Proc Natl Acad Sci U S A. 1988 Dec;85(24):9683–9687. doi: 10.1073/pnas.85.24.9683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Houng H. S., Kopecko D. J., Baron L. S. Molecular cloning and physical and functional characterization of the Salmonella typhimurium and Salmonella typhi galactose utilization operons. J Bacteriol. 1990 Aug;172(8):4392–4398. doi: 10.1128/jb.172.8.4392-4398.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hove-Jensen B., Nygaard P. Role of guanosine kinase in the utilization of guanosine for nucleotide synthesis in Escherichia coli. J Gen Microbiol. 1989 May;135(5):1263–1273. doi: 10.1099/00221287-135-5-1263. [DOI] [PubMed] [Google Scholar]
  11. Jennings M. P., van der Ley P., Wilks K. E., Maskell D. J., Poolman J. T., Moxon E. R. Cloning and molecular analysis of the galE gene of Neisseria meningitidis and its role in lipopolysaccharide biosynthesis. Mol Microbiol. 1993 Oct;10(2):361–369. [PubMed] [Google Scholar]
  12. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  13. Maskell D. J., Szabo M. J., Butler P. D., Williams A. E., Moxon E. R. Molecular analysis of a complex locus from Haemophilus influenzae involved in phase-variable lipopolysaccharide biosynthesis. Mol Microbiol. 1991 May;5(5):1013–1022. doi: 10.1111/j.1365-2958.1991.tb01874.x. [DOI] [PubMed] [Google Scholar]
  14. Maskell D. J., Szabo M. J., Deadman M. E., Moxon E. R. The gal locus from Haemophilus influenzae: cloning, sequencing and the use of gal mutants to study lipopolysaccharide. Mol Microbiol. 1992 Oct;6(20):3051–3063. doi: 10.1111/j.1365-2958.1992.tb01763.x. [DOI] [PubMed] [Google Scholar]
  15. Metzger M., Bellemann P., Bugert P., Geider K. Genetics of galactose metabolism of Erwinia amylovora and its influence on polysaccharide synthesis and virulence of the fire blight pathogen. J Bacteriol. 1994 Jan;176(2):450–459. doi: 10.1128/jb.176.2.450-459.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miller V. L., Farmer J. J., 3rd, Hill W. E., Falkow S. The ail locus is found uniquely in Yersinia enterocolitica serotypes commonly associated with disease. Infect Immun. 1989 Jan;57(1):121–131. doi: 10.1128/iai.57.1.121-131.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miller V. L., Mekalanos J. J. A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J Bacteriol. 1988 Jun;170(6):2575–2583. doi: 10.1128/jb.170.6.2575-2583.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Morona R., van den Bosch L., Manning P. A. Molecular, genetic, and topological characterization of O-antigen chain length regulation in Shigella flexneri. J Bacteriol. 1995 Feb;177(4):1059–1068. doi: 10.1128/jb.177.4.1059-1068.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Peng H. L., Fu T. F., Liu S. F., Chang H. Y. Cloning and expression of the Klebsiella pneumoniae galactose operon. J Biochem. 1992 Nov;112(5):604–608. doi: 10.1093/oxfordjournals.jbchem.a123947. [DOI] [PubMed] [Google Scholar]
  20. Pierson D. E., Falkow S. The ail gene of Yersinia enterocolitica has a role in the ability of the organism to survive serum killing. Infect Immun. 1993 May;61(5):1846–1852. doi: 10.1128/iai.61.5.1846-1852.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pierson D. E. Mutations affecting lipopolysaccharide enhance ail-mediated entry of Yersinia enterocolitica into mammalian cells. J Bacteriol. 1994 Jul;176(13):4043–4051. doi: 10.1128/jb.176.13.4043-4051.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Portnoy D. A., Moseley S. L., Falkow S. Characterization of plasmids and plasmid-associated determinants of Yersinia enterocolitica pathogenesis. Infect Immun. 1981 Feb;31(2):775–782. doi: 10.1128/iai.31.2.775-782.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Redfield R. J., Campbell A. M. Origin of cryptic lambda prophages in Escherichia coli K-12. Cold Spring Harb Symp Quant Biol. 1984;49:199–206. doi: 10.1101/sqb.1984.049.01.023. [DOI] [PubMed] [Google Scholar]
  24. Reeves P. Role of O-antigen variation in the immune response. Trends Microbiol. 1995 Oct;3(10):381–386. doi: 10.1016/s0966-842x(00)88983-0. [DOI] [PubMed] [Google Scholar]
  25. Robertson B. D., Frosch M., van Putten J. P. The role of galE in the biosynthesis and function of gonococcal lipopolysaccharide. Mol Microbiol. 1993 May;8(5):891–901. doi: 10.1111/j.1365-2958.1993.tb01635.x. [DOI] [PubMed] [Google Scholar]
  26. Schnaitman C. A., Klena J. D. Genetics of lipopolysaccharide biosynthesis in enteric bacteria. Microbiol Rev. 1993 Sep;57(3):655–682. doi: 10.1128/mr.57.3.655-682.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Skurnik M., Venho R., Toivanen P., al-Hendy A. A novel locus of Yersinia enterocolitica serotype O:3 involved in lipopolysaccharide outer core biosynthesis. Mol Microbiol. 1995 Aug;17(3):575–594. doi: 10.1111/j.1365-2958.1995.mmi_17030575.x. [DOI] [PubMed] [Google Scholar]
  28. Stevenson G., Kessler A., Reeves P. R. A plasmid-borne O-antigen chain length determinant and its relationship to other chain length determinants. FEMS Microbiol Lett. 1995 Jan 1;125(1):23–30. doi: 10.1111/j.1574-6968.1995.tb07330.x. [DOI] [PubMed] [Google Scholar]
  29. Stroeher U. H., Karageorgos L. E., Brown M. H., Morona R., Manning P. A. A putative pathway for perosamine biosynthesis is the first function encoded within the rfb region of Vibrio cholerae O1. Gene. 1995 Dec 1;166(1):33–42. doi: 10.1016/0378-1119(95)00589-0. [DOI] [PubMed] [Google Scholar]
  30. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  31. Wartenberg K., Lysy J., Knapp W. On the sugar content of the lipopolysaccharides of the various strains known as Yersinia enterocolitica. Zentralbl Bakteriol Orig A. 1975;230(3):361–366. [PubMed] [Google Scholar]
  32. Whitfield C. Biosynthesis of lipopolysaccharide O antigens. Trends Microbiol. 1995 May;3(5):178–185. doi: 10.1016/s0966-842x(00)88917-9. [DOI] [PubMed] [Google Scholar]
  33. Whitfield C., Valvano M. A. Biosynthesis and expression of cell-surface polysaccharides in gram-negative bacteria. Adv Microb Physiol. 1993;35:135–246. doi: 10.1016/s0065-2911(08)60099-5. [DOI] [PubMed] [Google Scholar]
  34. Winberg G., Hammarskjöld M. L. Isolation of DNA from agarose gels using DEAE-paper. Application to restriction site mapping of adenovirus type 16 DNA. Nucleic Acids Res. 1980 Jan 25;8(2):253–264. doi: 10.1093/nar/8.2.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zhang L., Toivanen P., Skurnik M. The gene cluster directing O-antigen biosynthesis in Yersinia enterocolitica serotype 0:8: identification of the genes for mannose and galactose biosynthesis and the gene for the O-antigen polymerase. Microbiology. 1996 Feb;142(Pt 2):277–288. doi: 10.1099/13500872-142-2-277. [DOI] [PubMed] [Google Scholar]
  36. al-Hendy A., Toivanen P., Skurnik M. The effect of growth temperature on the biosynthesis of Yersinia enterocolitica O:3 lipopolysaccharide: temperature regulates the transcription of the rfb but not of the rfa region. Microb Pathog. 1991 Jan;10(1):81–86. doi: 10.1016/0882-4010(91)90068-l. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES