Structure and Functional Genomics of Lipopolysaccharide Expression inHaemophilus Influenzae

  • James C. Richards
  • Andrew D. Cox
  • Elke K. H. Schweda
  • Adele Martin
  • Derek W. Hood
  • E. Richard Moxon
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 491)


The involvement of genes in the lic loci in H. influenzae LPS expression has been known for some time (20). However, it was not until recently that it was shown that the licl locus contains genes required for phase variable expression of phosphocholine substituents (21), while genes in the lic2 locus and lgtC are required for expression of the globoside trisaccharide, α-D-Galp-(1→4)- ß-D-Galp-(1→4)- ß-D-Glcp (i.e., the pK blood group epitope) (19,23). The availability of the complete sequence of the H. influenzae strain Rd genome has facilitated significant progress in understanding the role of these and other genes in the expression and biosynthesis of LPS. We have employed a comparative structural fingerprinting strategy to establish the structural relationships among LPS from H. influenzae mutant strains in which putative biosynthesis genes were inactivated. Using this functional genomics approach, we have gained considerable insight into the genetic basis for intra-strain and strain-to-strain variation in epitope expression.


Sialic Acid Chain Extension Neisseria Gonorrhoeae Oligosaccharide Chain Molecular Environment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Preston, A., Mandrell, R.E., Gibson, B.W., & Apicella, M.A. (1996) The lipopolysaccharide of Gram negative bacteria. Crit. Rev. Microbiol. 22: 139–1802.Google Scholar
  2. 2.
    Phillips, N.J., Apicella, M.A., Griffiss, J.M. & Gibson, B.W. (1992) Structural characterization of the cell surface lipooligosaccharides from a nontypable strain ofHaemophilus influenzae. Biochemistry31: 4515–4526.CrossRefGoogle Scholar
  3. 3.
    Schweda, E.K.H., Hegedus, O.E., Borelli, S., Lindberg, A.A.,Weiser. J.N.,Maskell, D.J.&Moxon, E.R. (1993) Structural studies of the cell envelope lipopolysacchaide fromHaemophilus influenzaestrain AH1–3 (lic3+).Carbohydr. Res.246: 319–330.PubMedCrossRefGoogle Scholar
  4. 4.
    Schweda E.K.H., Jansson, P.-E., Moxon, E.R. & Lindberg, A.A. (1995) Structural studies of the saccharide part of the cell envelope lipopolysaccharide fromHaemophilus influenzaestrain galEgalK.Carbohydr. Res.272: 213–224.PubMedCrossRefGoogle Scholar
  5. 5.
    Philips, N.J., McLaughlin, R., Miller, T.J., Apicella, M.A. & Gibson, B.W. (1996) Characterization of two transposon mutants fromHaemophilus influenzaetype b with altered lipooligosaccharide biosynthesis.Biochemistry35: 5937–5947.CrossRefGoogle Scholar
  6. 6.
    Masoud, H., Moxon, E.R., Martin, A., Krajcarski, D. & Richards, J.C. (1997) Structure of the variable and conserved lipopolysaccharide oligosaccharide epitopes expressed byHaemophilus influenzaeserotype b strain Eagan.Biochemistry36: 2091–2103.PubMedCrossRefGoogle Scholar
  7. 7.
    Risberg, A., Schweda, E.K.H & Jansson, P.-E. (1997) Structural studies of the cell-envelope oligosaccharide from the lipopolysaccharide ofHaemophilus influenzaestrain RM.118–28.Eur. J. Biochem.243: 701–707.PubMedCrossRefGoogle Scholar
  8. 8.
    Risberg, A., Masoud, H., Martin, A., Richards, J.C., Moxon, E.R. & Schweda E.K.H. (1999) Structural analysis of the lipopolysaccharide oligosaccharide epitopes expressed by a capsule-deficient strain ofHaemophilus influenzaeRd.Eur. J. Biochem.261: 717–180.CrossRefGoogle Scholar
  9. 9.
    Rahman, M.M., Gu, X.-X., Tsai, C.-M., Kolli, V.S.K & Carlson, R.W. (1999) The structural heterogeneity of lipooligosaccharide (LOS) expressed by pathogenic non-typeableHaemophilus influenzaestrain NTHi 9274.Glycobiology9: 1371–1380.PubMedCrossRefGoogle Scholar
  10. 10.
    Thibault, P. & Richards, J.C. (1999) Application of combined capillary electrophoresis electrospray mass spectrometry in the characterization of short-chain lipopolysaccharide:Haemophilus influenzae.InMethods in Molecular BiologyVol. 145: Bacterial Toxins: Methods and Protocols (Holst, O., ed.), pp. 327–344. Humana Press Inc., Totowa, NJ, USA.Google Scholar
  11. 11.
    Schweda, E.K.H., Brisson, J.-R., Alvelius, G., Martin, A., Weisser, J.N., Hood, D.W., Moxon, E.R. & Richards, J.C. (2000) Characterization of the phosphocholine-substituted oligosaccharide in lipopolysaccharides of type bHaemophilus influenzae. Eur. J Biochem.267: 3902–3913.CrossRefGoogle Scholar
  12. 12.
    Risberg, A., Alvelius, G., and Schweda, E.K.H. (1999) Structural analysis of the lipopolysaccharide oligosaccharide epitopes expressed byHaemophilus influenzaestrain RM.118–26.Eur. J. Biochem.265: 1067–1074.PubMedCrossRefGoogle Scholar
  13. 13.
    Hood, D.W., Makepeace, K., Deadman, M.E., Rest, R.F., Thibault, P., Martin, A., Richards, J.C., and Moxon, E.R. (1999) Sialic acid in the lipopolysaccharide ofHaemophilus influenzae:strain distribution, influence on serum resistance and structural characterisation.Mol. Microbio!.33: 679–692.CrossRefGoogle Scholar
  14. 14.
    Lysenko, E., Richards, J.C., Cox, A.D., Stewart, S., Martin, A., Kapoor, M. & Weiser, J.N. (2000) The position of phosphocholine on the lipopolysaccharide ofHaemophilus influenzaeaffects binding and sensitivity to C-reactive protein mediated killing.Mol. Microbio!.35: 234–245.CrossRefGoogle Scholar
  15. 15.
    Vimr, E., Lichtensteiger, C., and Steenbergen, S. (2000) Sialic acid metabolism’s dual function inHaemophilus influenzae. Mol. Microbiol.36: 1113–1123.CrossRefGoogle Scholar
  16. 16.
    Hood, D.W., Cox, A.D., Gilbert, M., Makepeace, K., Walsh, S., Deadman, M.E., Cody, A., Martin, A., Manson, M., Schweda, E.K.H., Brisson, J.-R., Richards, J.C., Moxon, E.R. & Wakarchuk, W.W. (2000) Identification of lipopolysaccharide a-2,3-sialyltranferase fromHaemophilus influenzae. Mol Microbiol.In Press.Google Scholar
  17. 17.
    Fleishmann, R.D., Adams, M.D., White, O., Clayton, R.A., Kirkness, E.F., Kerlavage, A.R., Butt, C.J., Tomb, F.-F., Doughtery, B.A, Merrick„ J.M., McKenney, K., Sutton, G., Fitzhugh, W., Fields, C., Gocaybe, J.D., Scott, J., Shirley, R., Liu, L.-I., Glodek, A., Kelly, J.M., Weildman, J.F., Phillips, C.A., Spiggs, T., Hedblom, E., Cotton, M.D., Utterbask, T.R., Hanna, M.C., Nguyen, D.T., Saudek, D.M., Brandon, R.C., Fine, L.D., Fritchmann, J.L., Georghagen, N.S.M., Gnehm, C.L., McDonald, L.A., Small, K.V., Fraser, C.M., Smith, H.O. & Venter, J.C. (1995) The genome ofHaemophilus influenzaeRd. Science 269: 496–512.CrossRefGoogle Scholar
  18. 18.
    Kimura, A. & Hansen, E.J. (1986) Antigenic and phenotypic variations ofHaemophilus influenzaetype b lipopolysaccharide and their relationship to virulence.Infect. Immun.51: 69–79.PubMedGoogle Scholar
  19. 19.
    Hood, D.W., Deadman, M.E., Allen, T., Masoud, H., Martin, A., Brisson, J.-R., Fleischmann, R., Venter, J.C., Richards, J.C., and Moxon, E.R. (1996) Use of the complete genome sequence information ofHaemophilus influenzaestrain Rd to investigate lipopolysaccharide biosynthesis.Mo! Microbio!22: 651–965.Google Scholar
  20. 20.
    Hood, D.W., and Moxon, E.R. (1999) Lipopolysaccharide phase variation inHaemophilusandNeisseria.InEndotoxin in Health and Disease.New York, USA: Marcel Dekker Inc,pp 39–54.Google Scholar
  21. 21.
    Weiser, J.N., Shchepetov, M. & Chong, S.T.H. (1997) Decoration of lipopolysaccharide with phosphocholine: a phase-variable characteristic ofHaemophilus influenzae. Infect. Immunity65: 943–950.Google Scholar
  22. 22.
    Banerjee, A., Wang, R., Uljon, S.E., Rice, R.A., Gotschlich, E.C., Stein, D.C., (1998) Identification of the gene(lgtG)encoding the lipooligosaccharide ß chain synthesizing glucosyltransferase fromNeisseria gonorrhoeae. Proc. Natl. Acad. Sci. USA 95:10872–10877.CrossRefGoogle Scholar
  23. 23.
    Hood, D.W., Cox, A.D., Wakarchuk, W.W., Schur, M., Schweda, E.K.H., Walsh, S., Deadman, M.E., Martin, A., Moxon, E.R., Richards, J.C., (2000) Genetic basis for expression of the major globotetraose containing lipopolysaccharide inHaemophilus influenzaestrain Rd (RM118).Submitted. Google Scholar
  24. 24.
    High, N.J., Deadman, M.E., and Moxon, E.R. (1993) The role of the repetitive DNA motif (5’-CAAT-3’) in the variable expression of theHaemophilus influenzaelipopolysaccharide epitope aGal (1–4)13Gal.Mol. Microbio!.9: 1275–1282.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2001

Authors and Affiliations

  • James C. Richards
    • 1
  • Andrew D. Cox
    • 1
  • Elke K. H. Schweda
    • 2
  • Adele Martin
    • 1
  • Derek W. Hood
    • 3
  • E. Richard Moxon
    • 3
  1. 1.National Research CouncilInstitute for Biological SciencesOttawa
  2. 2.Clinical Research CentreUniversity College of South StockholmHuddingeSweden
  3. 3.Molecular Infectious Diseases Group, University of Oxford, Department of PaediatricsJohn Radcliffe Hospital, HeadingtonUnited Kingdom

Personalised recommendations