Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Jun;178(11):3339–3341. doi: 10.1128/jb.178.11.3339-3341.1996

Molecular cloning of the Haemophilus influenzae gmhA (lpcA) gene encoding a phosphoheptose isomerase required for lipooligosaccharide biosynthesis.

J S Brooke 1, M A Valvano 1
PMCID: PMC178089  PMID: 8655517

Abstract

We have determined that gene HI#1181 of Haemophilus influenzae is a homolog of Escherichia coli gmhA (previously designated lpcA) (J. S. Brooke and M. A. Valvano, J. Biol. Chem. 271:3608-3614, 1996), which encodes a phosphoheptose isomerase catalyzing the first step of the biosynthesis of ADP-L-glycero-D-manno heptose. Mutations in this gene are associated with a heptoseless core lipopolysaccharide which determines an increased outer membrane permeability to hydrophobic compounds. The cloned H. influenzae gmhA restored the synthesis of a complete core in the gmhA-deleted E. coli strain chi711. Amino acid sequence comparisons of the GmhA proteins of E. coli and H. influenzae with other proteins in the databases revealed the existence of a novel family of phosphosugar a1do-keto isomerases.

Full Text

The Full Text of this article is available as a PDF (289.2 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Borrelli S., Hegedus O., Shaw D. H., Jansson P. E., Lindberg A. A. The tetrasaccharide L-alpha-D-heptose1-->2-L-alpha-D-heptose1--> 3-L-alpha-D-heptose1-->(3-deoxy-D-manno-octulosonic acid) and phosphate in lipid A define the conserved epitope in Haemophilus lipopolysaccharides recognized by a monoclonal antibody. Infect Immun. 1995 Sep;63(9):3683–3692. doi: 10.1128/iai.63.9.3683-3692.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brooke J. S., Valvano M. A. Biosynthesis of inner core lipopolysaccharide in enteric bacteria identification and characterization of a conserved phosphoheptose isomerase. J Biol Chem. 1996 Feb 16;271(7):3608–3614. doi: 10.1074/jbc.271.7.3608. [DOI] [PubMed] [Google Scholar]
  4. Cope L. D., Yogev R., Mertsola J., Argyle J. C., McCracken G. H., Jr, Hansen E. J. Effect of mutations in lipooligosaccharide biosynthesis genes on virulence of Haemophilus influenzae type b. Infect Immun. 1990 Jul;58(7):2343–2351. doi: 10.1128/iai.58.7.2343-2351.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Curtiss R., 3rd, Charamella L. J., Stallions D. R., Mays J. A. Parental functions during conjugation in Escherichia coli K-12. Bacteriol Rev. 1968 Dec;32(4 Pt 1):320–348. [PMC free article] [PubMed] [Google Scholar]
  6. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eidels L., Osborn M. J. Phosphoheptose isomerase, first enzyme in the biosynthesis of aldoheptose in Salmonella typhimurium. J Biol Chem. 1974 Sep 10;249(17):5642–5648. [PubMed] [Google Scholar]
  8. Fleischmann R. D., Adams M. D., White O., Clayton R. A., Kirkness E. F., Kerlavage A. R., Bult C. J., Tomb J. F., Dougherty B. A., Merrick J. M. Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science. 1995 Jul 28;269(5223):496–512. doi: 10.1126/science.7542800. [DOI] [PubMed] [Google Scholar]
  9. Helander I. M., Lindner B., Brade H., Altmann K., Lindberg A. A., Rietschel E. T., Zähringer U. Chemical structure of the lipopolysaccharide of Haemophilus influenzae strain I-69 Rd-/b+. Description of a novel deep-rough chemotype. Eur J Biochem. 1988 Nov 15;177(3):483–492. doi: 10.1111/j.1432-1033.1988.tb14398.x. [DOI] [PubMed] [Google Scholar]
  10. Kaneko T., Tanaka A., Sato S., Kotani H., Sazuka T., Miyajima N., Sugiura M., Tabata S. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. I. Sequence features in the 1 Mb region from map positions 64% to 92% of the genome. DNA Res. 1995 Aug 31;2(4):153-66, 191-8. doi: 10.1093/dnares/2.4.153. [DOI] [PubMed] [Google Scholar]
  11. Kimura A., Hansen E. J. Antigenic and phenotypic variations of Haemophilus influenzae type b lipopolysaccharide and their relationship to virulence. Infect Immun. 1986 Jan;51(1):69–79. doi: 10.1128/iai.51.1.69-79.1986. [DOI] [PMC free article] [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. Lee N. G., Sunshine M. G., Apicella M. A. Molecular cloning and characterization of the nontypeable Haemophilus influenzae 2019 rfaE gene required for lipopolysaccharide biosynthesis. Infect Immun. 1995 Mar;63(3):818–824. doi: 10.1128/iai.63.3.818-824.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Moxon E. R., Vaughn K. A. The type b capsular polysaccharide as a virulence determinant of Haemophilus influenzae: studies using clinical isolates and laboratory transformants. J Infect Dis. 1981 Apr;143(4):517–524. doi: 10.1093/infdis/143.4.517. [DOI] [PubMed] [Google Scholar]
  15. Peschke U., Schmidt H., Zhang H. Z., Piepersberg W. Molecular characterization of the lincomycin-production gene cluster of Streptomyces lincolnensis 78-11. Mol Microbiol. 1995 Jun;16(6):1137–1156. doi: 10.1111/j.1365-2958.1995.tb02338.x. [DOI] [PubMed] [Google Scholar]
  16. Phillips N. J., Apicella M. A., Griffiss J. M., Gibson B. W. Structural studies of the lipooligosaccharides from Haemophilus influenzae type b strain A2. Biochemistry. 1993 Mar 2;32(8):2003–2012. doi: 10.1021/bi00059a017. [DOI] [PubMed] [Google Scholar]
  17. Preston A., Maskell D., Johnson A., Moxon E. R. Altered lipopolysaccharide characteristic of the I69 phenotype in Haemophilus influenzae results from mutations in a novel gene, isn. J Bacteriol. 1996 Jan;178(2):396–402. doi: 10.1128/jb.178.2.396-402.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Qureshi N., Takayama K., Mascagni P., Honovich J., Wong R., Cotter R. J. Complete structural determination of lipopolysaccharide obtained from deep rough mutant of Escherichia coli. Purification by high performance liquid chromatography and direct analysis by plasma desorption mass spectrometry. J Biol Chem. 1988 Aug 25;263(24):11971–11976. [PubMed] [Google Scholar]
  19. Redfield R. J. sxy-1, a Haemophilus influenzae mutation causing greatly enhanced spontaneous competence. J Bacteriol. 1991 Sep;173(18):5612–5618. doi: 10.1128/jb.173.18.5612-5618.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rose I. A. Mechanism of the aldose-ketose isomerase reactions. Adv Enzymol Relat Areas Mol Biol. 1975;43:491–517. doi: 10.1002/9780470122884.ch6. [DOI] [PubMed] [Google Scholar]
  21. Schweda E. K., Sundström A. C., Eriksson L. M., Jonasson J. A., Lindberg A. A. Structural studies of the cell envelope lipopolysaccharides from Haemophilus ducreyi strains ITM 2665 and ITM 4747. J Biol Chem. 1994 Apr 22;269(16):12040–12048. [PubMed] [Google Scholar]
  22. Smith H. O., Tomb J. F., Dougherty B. A., Fleischmann R. D., Venter J. C. Frequency and distribution of DNA uptake signal sequences in the Haemophilus influenzae Rd genome. Science. 1995 Jul 28;269(5223):538–540. doi: 10.1126/science.7542802. [DOI] [PubMed] [Google Scholar]
  23. Tamaki S., Sato T., Matsuhashi M. Role of lipopolysaccharides in antibiotic resistance and bacteriophage adsorption of Escherichia coli K-12. J Bacteriol. 1971 Mar;105(3):968–975. doi: 10.1128/jb.105.3.968-975.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Turk D. C. The pathogenicity of Haemophilus influenzae. J Med Microbiol. 1984 Aug;18(1):1–16. doi: 10.1099/00222615-18-1-1. [DOI] [PubMed] [Google Scholar]
  25. Yao Z., Liu H., Valvano M. A. Acetylation of O-specific lipopolysaccharides from Shigella flexneri 3a and 2a occurs in Escherichia coli K-12 carrying cloned S. flexneri 3a and 2a rfb genes. J Bacteriol. 1992 Dec;174(23):7500–7508. doi: 10.1128/jb.174.23.7500-7508.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zwahlen A., Rubin L. G., Connelly C. J., Inzana T. J., Moxon E. R. Alteration of the cell wall of Haemophilus influenzae type b by transformation with cloned DNA: association with attenuated virulence. J Infect Dis. 1985 Sep;152(3):485–492. doi: 10.1093/infdis/152.3.485. [DOI] [PubMed] [Google Scholar]
  27. Zwahlen A., Rubin L. G., Moxon E. R. Contribution of lipopolysaccharide to pathogenicity of Haemophilus influenzae: comparative virulence of genetically-related strains in rats. Microb Pathog. 1986 Oct;1(5):465–473. doi: 10.1016/0882-4010(86)90008-2. [DOI] [PubMed] [Google Scholar]

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

RESOURCES