Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Nov;178(21):6281–6287. doi: 10.1128/jb.178.21.6281-6287.1996

Characterization of the genetic locus encoding Haemophilus influenzae type b surface fibrils.

J W St Geme 3rd 1, D Cutter 1, S J Barenkamp 1
PMCID: PMC178501  PMID: 8892830

Abstract

Haemophilus influenzae is a common gram-negative pathogen that initiates infection by colonizing the upper respiratory tract epithelium. In previous work, we reported the isolation of a locus involved in expression of short, thin surface fibrils by H. influenzae type b and presented evidence that surface fibrils promote attachment to human epithelial cells. In the present study, we determined that the fibril locus is composed of one long open reading frame, designated hsf, which encodes a protein (Hsf) with a molecular mass of approximately 240 kDa. The derived amino acid sequence of the hsf product demonstrated 81% similarity and 72% identity to a recently identified nontypeable H. influenzae adhesin referred to as Hia. In experiments with a panel of eight cultured cell lines, the Hsf and Hia proteins were found to confer the same binding specificities, suggesting that hsf and hia are alleles of the same locus. Southern analysis and mutagenesis studies reinforced this conclusion. Further investigation revealed that an hsf homolog is ubiquitous among encapsulated H. influenzae strains and is present in a subset of nontypeable Haemophilus strains as well. We speculate that the hsf gene product plays an important role in the process of respiratory tract colonization by H. influenzae.

Full Text

The Full Text of this article is available as a PDF (720.7 KB).

Selected References

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

  1. ALEXANDER H. E., LEIDY G. Determination of inherited traits of H. influenzae by desoxyribonucleic acid fractions isolated from type-specific cells. J Exp Med. 1951 Apr 1;93(4):345–359. doi: 10.1084/jem.93.4.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Anderson P., Johnston R. B., Jr, Smith D. H. Human serum activities against Hemophilus influenzae, type b. J Clin Invest. 1972 Jan;51(1):31–38. doi: 10.1172/JCI106793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barenkamp S. J., Leininger E. Cloning, expression, and DNA sequence analysis of genes encoding nontypeable Haemophilus influenzae high-molecular-weight surface-exposed proteins related to filamentous hemagglutinin of Bordetella pertussis. Infect Immun. 1992 Apr;60(4):1302–1313. doi: 10.1128/iai.60.4.1302-1313.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barenkamp S. J., St Geme J. W., 3rd Genes encoding high-molecular-weight adhesion proteins of nontypeable Haemophilus influenzae are part of gene clusters. Infect Immun. 1994 Aug;62(8):3320–3328. doi: 10.1128/iai.62.8.3320-3328.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Barenkamp S. J., St Geme J. W., 3rd Identification of a second family of high-molecular-weight adhesion proteins expressed by non-typable Haemophilus influenzae. Mol Microbiol. 1996 Mar;19(6):1215–1223. doi: 10.1111/j.1365-2958.1996.tb02467.x. [DOI] [PubMed] [Google Scholar]
  7. Benjelloun-Touimi Z., Sansonetti P. J., Parsot C. SepA, the major extracellular protein of Shigella flexneri: autonomous secretion and involvement in tissue invasion. Mol Microbiol. 1995 Jul;17(1):123–135. doi: 10.1111/j.1365-2958.1995.mmi_17010123.x. [DOI] [PubMed] [Google Scholar]
  8. Benz I., Schmidt M. A. AIDA-I, the adhesin involved in diffuse adherence of the diarrhoeagenic Escherichia coli strain 2787 (O126:H27), is synthesized via a precursor molecule. Mol Microbiol. 1992 Jun;6(11):1539–1546. doi: 10.1111/j.1365-2958.1992.tb00875.x. [DOI] [PubMed] [Google Scholar]
  9. Brinton C. C., Jr, Carter M. J., Derber D. B., Kar S., Kramarik J. A., To A. C., To S. C., Wood S. W. Design and development of pilus vaccines for Haemophilus influenzae diseases. Pediatr Infect Dis J. 1989 Jan;8(1 Suppl):S54–S61. [PubMed] [Google Scholar]
  10. Cochi S. L., Broome C. V. Vaccine prevention of Haemophilus influenzae type b disease: past, present and future. Pediatr Infect Dis. 1986 Jan-Feb;5(1):12–19. doi: 10.1097/00006454-198601000-00003. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Dower W. J., Miller J. F., Ragsdale C. W. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988 Jul 11;16(13):6127–6145. doi: 10.1093/nar/16.13.6127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Farley M. M., Stephens D. S., Kaplan S. L., Mason E. O., Jr Pilus- and non-pilus-mediated interactions of Haemophilus influenzae type b with human erythrocytes and human nasopharyngeal mucosa. J Infect Dis. 1990 Feb;161(2):274–280. doi: 10.1093/infdis/161.2.274. [DOI] [PubMed] [Google Scholar]
  14. Farley M. M., Stephens D. S., Mulks M. H., Cooper M. D., Bricker J. V., Mirra S. S., Wright A. Pathogenesis of IgA1 protease-producing and -nonproducing Haemophilus influenzae in human nasopharyngeal organ cultures. J Infect Dis. 1986 Nov;154(5):752–759. doi: 10.1093/infdis/154.5.752. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Geme J. W., 3rd, Cutter D. Evidence that surface fibrils expressed by Haemophilus influenzae type b promote attachment to human epithelial cells. Mol Microbiol. 1995 Jan;15(1):77–85. doi: 10.1111/j.1365-2958.1995.tb02222.x. [DOI] [PubMed] [Google Scholar]
  17. Gilsdorf J. R., Chang H. Y., McCrea K. W., Bakaletz L. O. Comparison of hemagglutinating pili of Haemophilus influenzae type b with similar structures of nontypeable H. influenzae. Infect Immun. 1992 Feb;60(2):374–379. doi: 10.1128/iai.60.2.374-379.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Guerina N. G., Langermann S., Clegg H. W., Kessler T. W., Goldman D. A., Gilsdorf J. R. Adherence of piliated Haemophilus influenzae type b to human oropharyngeal cells. J Infect Dis. 1982 Oct;146(4):564–564. doi: 10.1093/infdis/146.4.564. [DOI] [PubMed] [Google Scholar]
  19. Herriott R. M., Meyer E. M., Vogt M. Defined nongrowth media for stage II development of competence in Haemophilus influenzae. J Bacteriol. 1970 Feb;101(2):517–524. doi: 10.1128/jb.101.2.517-524.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kilian M. A taxonomic study of the genus Haemophilus, with the proposal of a new species. J Gen Microbiol. 1976 Mar;93(1):9–62. doi: 10.1099/00221287-93-1-9. [DOI] [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Loeb M. R., Connor E., Penney D. A comparison of the adherence of fimbriated and nonfimbriated Haemophilus influenzae type b to human adenoids in organ culture. Infect Immun. 1988 Feb;56(2):484–489. doi: 10.1128/iai.56.2.484-489.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Murphy T. F., Bernstein J. M., Dryja D. M., Campagnari A. A., Apicella M. A. Outer membrane protein and lipooligosaccharide analysis of paired nasopharyngeal and middle ear isolates in otitis media due to nontypable Haemophilus influenzae: pathogenetic and epidemiological observations. J Infect Dis. 1987 Nov;156(5):723–731. doi: 10.1093/infdis/156.5.723. [DOI] [PubMed] [Google Scholar]
  24. Musser J. M., Kroll J. S., Granoff D. M., Moxon E. R., Brodeur B. R., Campos J., Dabernat H., Frederiksen W., Hamel J., Hammond G. Global genetic structure and molecular epidemiology of encapsulated Haemophilus influenzae. Rev Infect Dis. 1990 Jan-Feb;12(1):75–111. doi: 10.1093/clinids/12.1.75. [DOI] [PubMed] [Google Scholar]
  25. Musser J. M., Kroll J. S., Moxon E. R., Selander R. K. Evolutionary genetics of the encapsulated strains of Haemophilus influenzae. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7758–7762. doi: 10.1073/pnas.85.20.7758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pichichero M. E., Loeb M., Anderson, Smith D. H. Do pili play a role in pathogenicity of Haemophilus influenzae type B? Lancet. 1982 Oct 30;2(8305):960–962. doi: 10.1016/s0140-6736(82)90161-1. [DOI] [PubMed] [Google Scholar]
  27. Provence D. L., Curtiss R., 3rd Isolation and characterization of a gene involved in hemagglutination by an avian pathogenic Escherichia coli strain. Infect Immun. 1994 Apr;62(4):1369–1380. doi: 10.1128/iai.62.4.1369-1380.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Read R. C., Wilson R., Rutman A., Lund V., Todd H. C., Brain A. P., Jeffery P. K., Cole P. J. Interaction of nontypable Haemophilus influenzae with human respiratory mucosa in vitro. J Infect Dis. 1991 Mar;163(3):549–558. doi: 10.1093/infdis/163.3.549. [DOI] [PubMed] [Google Scholar]
  29. Sable N. S., Connor E. M., Hall C. B., Loeb M. R. Variable adherence of fimbriated Haemophilus influenzae type b to human cells. Infect Immun. 1985 Apr;48(1):119–123. doi: 10.1128/iai.48.1.119-123.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. St Geme J. W., 3rd, Cutter D. Influence of pili, fibrils, and capsule on in vitro adherence by Haemophilus influenzae type b. Mol Microbiol. 1996 Jul;21(1):21–31. doi: 10.1046/j.1365-2958.1996.6241331.x. [DOI] [PubMed] [Google Scholar]
  31. St Geme J. W., 3rd, Falkow S., Barenkamp S. J. High-molecular-weight proteins of nontypable Haemophilus influenzae mediate attachment to human epithelial cells. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2875–2879. doi: 10.1073/pnas.90.7.2875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. St Geme J. W., 3rd, Falkow S. Haemophilus influenzae adheres to and enters cultured human epithelial cells. Infect Immun. 1990 Dec;58(12):4036–4044. doi: 10.1128/iai.58.12.4036-4044.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. St Geme J. W., 3rd, Falkow S. Loss of capsule expression by Haemophilus influenzae type b results in enhanced adherence to and invasion of human cells. Infect Immun. 1991 Apr;59(4):1325–1333. doi: 10.1128/iai.59.4.1325-1333.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. St Geme J. W., 3rd Nontypeable Haemophilus influenzae disease: epidemiology, pathogenesis, and prospects for prevention. Infect Agents Dis. 1993 Feb;2(1):1–16. [PubMed] [Google Scholar]
  35. St Geme J. W., 3rd, de la Morena M. L., Falkow S. A Haemophilus influenzae IgA protease-like protein promotes intimate interaction with human epithelial cells. Mol Microbiol. 1994 Oct;14(2):217–233. doi: 10.1111/j.1365-2958.1994.tb01283.x. [DOI] [PubMed] [Google Scholar]
  36. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  37. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  40. Walker J. E., Saraste M., Runswick M. J., Gay N. J. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J. 1982;1(8):945–951. doi: 10.1002/j.1460-2075.1982.tb01276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Weber A., Harris K., Lohrke S., Forney L., Smith A. L. Inability to express fimbriae results in impaired ability of Haemophilus influenzae b to colonize the nasopharynx. Infect Immun. 1991 Dec;59(12):4724–4728. doi: 10.1128/iai.59.12.4724-4728.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weiser J. N., Chong S. T., Greenberg D., Fong W. Identification and characterization of a cell envelope protein of Haemophilus influenzae contributing to phase variation in colony opacity and nasopharyngeal colonization. Mol Microbiol. 1995 Aug;17(3):555–564. doi: 10.1111/j.1365-2958.1995.mmi_17030555.x. [DOI] [PubMed] [Google Scholar]
  43. Willems R., Paul A., van der Heide H. G., ter Avest A. R., Mooi F. R. Fimbrial phase variation in Bordetella pertussis: a novel mechanism for transcriptional regulation. EMBO J. 1990 Sep;9(9):2803–2809. doi: 10.1002/j.1460-2075.1990.tb07468.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  45. Yogev D., Rosengarten R., Watson-McKown R., Wise K. S. Molecular basis of Mycoplasma surface antigenic variation: a novel set of divergent genes undergo spontaneous mutation of periodic coding regions and 5' regulatory sequences. EMBO J. 1991 Dec;10(13):4069–4079. doi: 10.1002/j.1460-2075.1991.tb04983.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES