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. 1996 Nov;178(22):6564–6570. doi: 10.1128/jb.178.22.6564-6570.1996

Duplication of pilus gene complexes of Haemophilus influenzae biogroup aegyptius.

T D Read 1, M Dowdell 1, S W Satola 1, M M Farley 1
PMCID: PMC178543  PMID: 8932313

Abstract

Brazilian purpuric fever (BPF) is a recently described pediatric septicemia caused by a strain of Haemophilus influenzae biogroup aegyptius. The pilus specified by this bacterium may be important in BPF pathogenesis, enhancing attachment to host tissue. Here, we report the cloning of two haf (for H. influenzae biogroup aegyptius fimbriae) gene clusters from a cosmid library of strain F3031. We sequenced a 6.8-kb segment of the haf1 cluster and identified five genes (hafA to hafE). The predicted protein products, HafA to HafD, are 72, 95, 98, and 90% similar, respectively, to HifA to HifD of the closely related H. influenzae type b pilus. Strikingly, the putative pilus adhesion, HifE, shares only 44% identity with HafE, suggesting that the proteins may differ in receptor specificity. Insertion of a mini-gammadelta transposon in the hafE gene eliminated hemadsorption. The nucleotide sequences of the haf1 and haf2 clusters are more than 99% identical. Using the recently published sequence of the H. influenzae Rd genome, we determined that the haf1 complex lies at a unique position in the chromosome between the pmbA gene and a hypothetical open reading frame, HI1153. The location of the haf2 cluster, inserted between the purE and pepN genes, is analogous to the hif genes on H. influenzae type b. BPF fimbrial phase switching appears to involve slip-strand mispairing of repeated dinucleotides in the pilus promoter. The BPF-associated H. influenzae biogroup aegyptius pilus system generally resembles other H. influenzae, but the possession of a second fimbrial gene cluster, which appears to have arisen by a recent duplication event, and the novel sequence of the HafE adhesin may be significant in the unusual pathogenesis of BPF.

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Selected References

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  1. Berg C. M., Wang G., Strausbaugh L. D., Berg D. E. Transposon-facilitated sequencing of DNAs cloned in plasmids. Methods Enzymol. 1993;218:279–306. doi: 10.1016/0076-6879(93)18022-5. [DOI] [PubMed] [Google Scholar]
  2. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  3. Collins J., Brüning H. J. Plasmids useable as gene-cloning vectors in an in vitro packaging by coliphage lambda: "cosmids". Gene. 1978 Oct;4(2):85–107. doi: 10.1016/0378-1119(78)90023-9. [DOI] [PubMed] [Google Scholar]
  4. Connor E. M., Loeb M. R. A hemadsorption method for detection of colonies of Haemophilus influenzae type b expressing fimbriae. J Infect Dis. 1983 Nov;148(5):855–860. doi: 10.1093/infdis/148.5.855. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Farley M. M., Whitney A. M., Spellman P., Quinn F. D., Weyant R. S., Mayer L., Stephens D. S. Analysis of the attachment and invasion of human epithelial cells by Haemophilus influenzae biogroup aegyptius. J Infect Dis. 1992 Jun;165 (Suppl 1):S111–S114. doi: 10.1093/infdis/165-supplement_1-s111. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Foglino M., Gharbi S., Lazdunski A. Nucleotide sequence of the pepN gene encoding aminopeptidase N of Escherichia coli. Gene. 1986;49(3):303–309. doi: 10.1016/0378-1119(86)90366-5. [DOI] [PubMed] [Google Scholar]
  9. Forney L. J., Marrs C. F., Bektesh S. L., Gilsdorf J. R. Comparison and analysis of the nucleotide sequences of pilin genes from Haemophilus influenzae type b strains Eagan and M43. Infect Immun. 1991 Jun;59(6):1991–1996. doi: 10.1128/iai.59.6.1991-1996.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gerlach G. F., Clegg S., Allen B. L. Identification and characterization of the genes encoding the type 3 and type 1 fimbrial adhesins of Klebsiella pneumoniae. J Bacteriol. 1989 Mar;171(3):1262–1270. doi: 10.1128/jb.171.3.1262-1270.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gilsdorf J. R., Marrs C. F., McCrea K. W., Forney L. J. Cloning, expression, and sequence analysis of the Haemophilus influenzae type b strain M43p+ pilin gene. Infect Immun. 1990 Apr;58(4):1065–1072. doi: 10.1128/iai.58.4.1065-1072.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hacker J. Genetic determinants coding for fimbriae and adhesins of extraintestinal Escherichia coli. Curr Top Microbiol Immunol. 1990;151:1–27. doi: 10.1007/978-3-642-74703-8_1. [DOI] [PubMed] [Google Scholar]
  13. Haslam D. B., Borén T., Falk P., Ilver D., Chou A., Xu Z., Normark S. The amino-terminal domain of the P-pilus adhesin determines receptor specificity. Mol Microbiol. 1994 Nov;14(3):399–409. doi: 10.1111/j.1365-2958.1994.tb02175.x. [DOI] [PubMed] [Google Scholar]
  14. Hultgren S. J., Lindberg F., Magnusson G., Kihlberg J., Tennent J. M., Normark S. The PapG adhesin of uropathogenic Escherichia coli contains separate regions for receptor binding and for the incorporation into the pilus. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4357–4361. doi: 10.1073/pnas.86.12.4357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hultgren S. J., Normark S. Biogenesis of the bacterial pilus. Curr Opin Genet Dev. 1991 Oct;1(3):313–318. doi: 10.1016/s0959-437x(05)80293-x. [DOI] [PubMed] [Google Scholar]
  16. Kar S., To S. C., Brinton C. C., Jr Cloning and expression in Escherichia coli of LKP pilus genes from a nontypeable Haemophilus influenzae strain. Infect Immun. 1990 Apr;58(4):903–908. doi: 10.1128/iai.58.4.903-908.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kuehn M. J., Ogg D. J., Kihlberg J., Slonim L. N., Flemmer K., Bergfors T., Hultgren S. J. Structural basis of pilus subunit recognition by the PapD chaperone. Science. 1993 Nov 19;262(5137):1234–1241. doi: 10.1126/science.7901913. [DOI] [PubMed] [Google Scholar]
  18. Langermann S., Wright A. Molecular analysis of the Haemophilus influenzae type b pilin gene. Mol Microbiol. 1990 Feb;4(2):221–230. doi: 10.1111/j.1365-2958.1990.tb00589.x. [DOI] [PubMed] [Google Scholar]
  19. Lindberg F. P., Lund B., Normark S. Genes of pyelonephritogenic E. coli required for digalactoside-specific agglutination of human cells. EMBO J. 1984 May;3(5):1167–1173. doi: 10.1002/j.1460-2075.1984.tb01946.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lintermans P. F., Bertels A., Schlicker C., Deboeck F., Charlier G., Pohl P., Norgren M., Normark S., van Montagu M., De Greve H. Identification, characterization, and nucleotide sequence of the F17-G gene, which determines receptor binding of Escherichia coli F17 fimbriae. J Bacteriol. 1991 Jun;173(11):3366–3373. doi: 10.1128/jb.173.11.3366-3373.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Marklund B. I., Tennent J. M., Garcia E., Hamers A., Båga M., Lindberg F., Gaastra W., Normark S. Horizontal gene transfer of the Escherichia coli pap and prs pili operons as a mechanism for the development of tissue-specific adhesive properties. Mol Microbiol. 1992 Aug;6(16):2225–2242. doi: 10.1111/j.1365-2958.1992.tb01399.x. [DOI] [PubMed] [Google Scholar]
  22. Mayer L. W., Bibb W. F., Birkness K. A., Irino K., Weyant R. S., Reeves M. W., Swenson J. M. Distinguishing clonal characteristics of the Brazilian purpuric fever-producing strain. The Brazilian Purpuric Fever Study Group. Pediatr Infect Dis J. 1989 Apr;8(4):241–243. [PubMed] [Google Scholar]
  23. McCrea K. W., Watson W. J., Gilsdorf J. R., Marrs C. F. Identification of hifD and hifE in the pilus gene cluster of Haemophilus influenzae type b strain Eagan. Infect Immun. 1994 Nov;62(11):4922–4928. doi: 10.1128/iai.62.11.4922-4928.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moxon E. R., Deich R. A., Connelly C. Cloning of chromosomal DNA from Haemophilus influenzae. Its use for studying the expression of type b capsule and virulence. J Clin Invest. 1984 Feb;73(2):298–306. doi: 10.1172/JCI111214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Porto M. H., Noel G. J., Edelson P. J. Resistance to serum bactericidal activity distinguishes Brazilian purpuric fever (BPF) case strains of Haemophilus influenzae biogroup aegyptius (H. aegyptius) from non-BPF strains. Brazilian Purpuric Fever Study Group. J Clin Microbiol. 1989 Apr;27(4):792–794. doi: 10.1128/jcm.27.4.792-794.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Quinn F. D., Weyant R. S., Worley M. J., White E. H., Utt E. A., Ades E. A. Human microvascular endothelial tissue culture cell model for studying pathogenesis of Brazilian purpuric fever. Infect Immun. 1995 Jun;63(6):2317–2322. doi: 10.1128/iai.63.6.2317-2322.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Robertson B. D., Meyer T. F. Genetic variation in pathogenic bacteria. Trends Genet. 1992 Dec;8(12):422–427. doi: 10.1016/0168-9525(92)90325-x. [DOI] [PubMed] [Google Scholar]
  28. Rodríguez-Sáinz M. C., Hernández-Chico C., Moreno F. Molecular characterization of pmbA, an Escherichia coli chromosomal gene required for the production of the antibiotic peptide MccB17. Mol Microbiol. 1990 Nov;4(11):1921–1932. doi: 10.1111/j.1365-2958.1990.tb02041.x. [DOI] [PubMed] [Google Scholar]
  29. Rubin L. G., Gloster E. S., Carlone G. M. An infant rat model of bacteremia with Brazilian purpuric fever isolates of Hemophilus influenzae biogroup aegyptius. Brazilian Purpuric Fever Study Group. J Infect Dis. 1989 Sep;160(3):476–482. doi: 10.1093/infdis/160.3.476. [DOI] [PubMed] [Google Scholar]
  30. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Smith T. F., Waterman M. S. Identification of common molecular subsequences. J Mol Biol. 1981 Mar 25;147(1):195–197. doi: 10.1016/0022-2836(81)90087-5. [DOI] [PubMed] [Google Scholar]
  32. St Geme J. W., 3rd, Falkow S. Isolation, expression, and nucleotide sequencing of the pilin structural gene of the Brazilian purpuric fever clone of Haemophilus influenzae biogroup aegyptius. Infect Immun. 1993 May;61(5):2233–2237. doi: 10.1128/iai.61.5.2233-2237.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. St Geme J. W., 3rd, Gilsdorf J. R., Falkow S. Surface structures and adherence properties of diverse strains of Haemophilus influenzae biogroup aegyptius. Infect Immun. 1991 Oct;59(10):3366–3371. doi: 10.1128/iai.59.10.3366-3371.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tiedeman A. A., Keyhani J., Kamholz J., Daum H. A., 3rd, Gots J. S., Smith J. M. Nucleotide sequence analysis of the purEK operon encoding 5'-phosphoribosyl-5-aminoimidazole carboxylase of Escherichia coli K-12. J Bacteriol. 1989 Jan;171(1):205–212. doi: 10.1128/jb.171.1.205-212.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Vieira J., Messing J. New pUC-derived cloning vectors with different selectable markers and DNA replication origins. Gene. 1991 Apr;100:189–194. doi: 10.1016/0378-1119(91)90365-i. [DOI] [PubMed] [Google Scholar]
  36. Watson W. J., Gilsdorf J. R., Tucci M. A., McCrea K. W., Forney L. J., Marrs C. F. Identification of a gene essential for piliation in Haemophilus influenzae type b with homology to the pilus assembly platform genes of gram-negative bacteria. Infect Immun. 1994 Feb;62(2):468–475. doi: 10.1128/iai.62.2.468-475.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weyant R. S., Bibb W. F., Stephens D. S., Holloway B. P., Moo-Penn W. F., Birkness K. A., Helsel L. O., Mayer L. W. Purification and characterization of a pilin specific for Brazilian purpuric fever-associated Haemophilus influenzae biogroup aegyptius (H. aegyptius) strains. J Clin Microbiol. 1990 Apr;28(4):756–763. doi: 10.1128/jcm.28.4.756-763.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weyant R. S., Quinn F. D., Utt E. A., Worley M., George V. G., Candal F. J., Ades E. W. Human microvascular endothelial cell toxicity caused by Brazilian purpuric fever-associated strains of Haemophilus influenzae biogroup aegyptius. J Infect Dis. 1994 Feb;169(2):430–433. doi: 10.1093/infdis/169.2.430. [DOI] [PubMed] [Google Scholar]
  39. Whitney A. M., Farley M. M. Cloning and sequence analysis of the structural pilin gene of Brazilian purpuric fever-associated Haemophilus influenzae biogroup aegyptius. Infect Immun. 1993 Apr;61(4):1559–1562. doi: 10.1128/iai.61.4.1559-1562.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Willems R. J., van der Heide H. G., Mooi F. R. Characterization of a Bordetella pertussis fimbrial gene cluster which is located directly downstream of the filamentous haemagglutinin gene. Mol Microbiol. 1992 Sep;6(18):2661–2671. doi: 10.1111/j.1365-2958.1992.tb01443.x. [DOI] [PubMed] [Google Scholar]
  41. Xu Z., Jones C. H., Haslam D., Pinkner J. S., Dodson K., Kihlberg J., Hultgren S. J. Molecular dissection of PapD interaction with PapG reveals two chaperone-binding sites. Mol Microbiol. 1995 Jun;16(5):1011–1020. doi: 10.1111/j.1365-2958.1995.tb02326.x. [DOI] [PubMed] [Google Scholar]
  42. Zav'yalov V. P., Zav'yalova G. A., Denesyuk A. I., Korpela T. Modelling of steric structure of a periplasmic molecular chaperone Caf1M of Yersinia pestis, a prototype member of a subfamily with characteristic structural and functional features. FEMS Immunol Med Microbiol. 1995 Mar;11(1):19–24. doi: 10.1111/j.1574-695X.1995.tb00074.x. [DOI] [PubMed] [Google Scholar]
  43. van Ham S. M., Mooi F. R., Sindhunata M. G., Maris W. R., van Alphen L. Cloning and expression in Escherichia coli of Haemophilus influenzae fimbrial genes establishes adherence to oropharyngeal epithelial cells. EMBO J. 1989 Nov;8(11):3535–3540. doi: 10.1002/j.1460-2075.1989.tb08519.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. van Ham S. M., van Alphen L., Mooi F. R., van Putten J. P. Phase variation of H. influenzae fimbriae: transcriptional control of two divergent genes through a variable combined promoter region. Cell. 1993 Jun 18;73(6):1187–1196. doi: 10.1016/0092-8674(93)90647-9. [DOI] [PubMed] [Google Scholar]
  45. van Ham S. M., van Alphen L., Mooi F. R., van Putten J. P. The fimbrial gene cluster of Haemophilus influenzae type b. Mol Microbiol. 1994 Aug;13(4):673–684. doi: 10.1111/j.1365-2958.1994.tb00461.x. [DOI] [PubMed] [Google Scholar]
  46. von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. doi: 10.1111/j.1432-1033.1983.tb07424.x. [DOI] [PubMed] [Google Scholar]

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