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. 1993 Jan;175(2):457–464. doi: 10.1128/jb.175.2.457-464.1993

Proteus mirabilis MR/P fimbriae: molecular cloning, expression, and nucleotide sequence of the major fimbrial subunit gene.

F K Bahrani 1, H L Mobley 1
PMCID: PMC196160  PMID: 8093447

Abstract

Proteus mirabilis, a cause of serious urinary tract infection and acute pyelonephritis, produces several putative virulence determinants, among them, fimbriae. Principally, two fimbrial types are produced by this species: mannose-resistant/Proteus-like (MR/P) fimbriae and mannose-resistant/Klebsiella-like (MR/K) fimbriae. To isolate MR/P fimbrial gene sequences, a P. mirabilis cosmid library was screened by immunoblotting and by hybridization with an oligonucleotide probe based on the N-terminal amino acid sequence of the isolated fimbrial polypeptide, ADQGHGTVKFVGSIIDAPCS. One clone, pMRP101, reacted strongly with a monoclonal antibody specific for MR/P fimbriae and with the DNA probe. This clone hemagglutinated both tannic acid-treated and untreated chicken erythrocytes with or without 50 mM D-mannose and was shown to be fimbriated by transmission electron microscopy. A 525-bp open reading frame, designated mrpA, predicted a 175-amino-acid polypeptide including a 23-amino-acid hydrophobic leader peptide. The unprocessed and processed polypeptides are predicted to be 17,909 and 15,689 Da, respectively. The N-terminal amino acid sequence of the processed fimbrial subunit exactly matched amino acid residues 24 to 43 predicted by the mrpA nucleotide sequence. The MrpA polypeptide shares 57% amino acid sequence identity with SmfA, the major fimbrial subunit of Serratia marcescens mannose-resistant fimbriae.

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Selected References

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

  1. Adegbola R. A., Old D. C., Senior B. W. The adhesins and fimbriae of Proteus mirabilis strains associated with high and low affinity for the urinary tract. J Med Microbiol. 1983 Nov;16(4):427–431. doi: 10.1099/00222615-16-4-427. [DOI] [PubMed] [Google Scholar]
  2. Adler H. I., Fisher W. D., Cohen A., Hardigree A. A. MINIATURE escherichia coli CELLS DEFICIENT IN DNA. Proc Natl Acad Sci U S A. 1967 Feb;57(2):321–326. doi: 10.1073/pnas.57.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bahrani F. K., Johnson D. E., Robbins D., Mobley H. L. Proteus mirabilis flagella and MR/P fimbriae: isolation, purification, N-terminal analysis, and serum antibody response following experimental urinary tract infection. Infect Immun. 1991 Oct;59(10):3574–3580. doi: 10.1128/iai.59.10.3574-3580.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Båga M., Normark S., Hardy J., O'Hanley P., Lark D., Olsson O., Schoolnik G., Falkow S. Nucleotide sequence of the papA gene encoding the Pap pilus subunit of human uropathogenic Escherichia coli. J Bacteriol. 1984 Jan;157(1):330–333. doi: 10.1128/jb.157.1.330-333.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Edén C. S., Larsson P., Lomberg H. Attachment of Proteus mirabilis to human urinary sediment epithelial cells in vitro is different from that of Escherichia coli. Infect Immun. 1980 Mar;27(3):804–807. doi: 10.1128/iai.27.3.804-807.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Elliott S. J., Nandapalan N., Chang B. J. Production of type 1 fimbriae by Escherichia coli HB101. Microb Pathog. 1991 Jun;10(6):481–486. doi: 10.1016/0882-4010(91)90114-p. [DOI] [PubMed] [Google Scholar]
  8. Gerlach G. F., Allen B. L., Clegg S. Molecular characterization of the type 3 (MR/K) fimbriae of Klebsiella pneumoniae. J Bacteriol. 1988 Aug;170(8):3547–3553. doi: 10.1128/jb.170.8.3547-3553.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gleckman R., Blagg N., Hibert D., Hall A., Crowley M., Pritchard A., Warren W. Symptomatic pyelonephritis in elderly men. J Am Geriatr Soc. 1982 Nov;30(11):690–693. doi: 10.1111/j.1532-5415.1982.tb01981.x. [DOI] [PubMed] [Google Scholar]
  10. Griffith D. P., Musher D. M., Itin C. Urease. The primary cause of infection-induced urinary stones. Invest Urol. 1976 Mar;13(5):346–350. [PubMed] [Google Scholar]
  11. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  12. Inouye M., Halegoua S. Secretion and membrane localization of proteins in Escherichia coli. CRC Crit Rev Biochem. 1980;7(4):339–371. doi: 10.3109/10409238009105465. [DOI] [PubMed] [Google Scholar]
  13. Jones B. D., Lockatell C. V., Johnson D. E., Warren J. W., Mobley H. L. Construction of a urease-negative mutant of Proteus mirabilis: analysis of virulence in a mouse model of ascending urinary tract infection. Infect Immun. 1990 Apr;58(4):1120–1123. doi: 10.1128/iai.58.4.1120-1123.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kiely B., Rees J. P. Sex differences in urinary tract infection in children. Ir Med J. 1984 Dec;77(12):384–387. [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Legnani-Fajardo C., Zunino P., Algorta G., Laborde H. F. Antigenic and immunogenic activity of flagella and fimbriae preparations from uropathogenic Proteus mirabilis. Can J Microbiol. 1991 Apr;37(4):325–328. doi: 10.1139/m91-052. [DOI] [PubMed] [Google Scholar]
  18. Lomberg H., Larsson P., Leffler H., Svanborg-Edén C. Different binding specificities of P. mirabilis compared to E. coli. Scand J Infect Dis Suppl. 1982;33:37–42. [PubMed] [Google Scholar]
  19. Mann P. G. Proteus urinary infections in childhood. J Clin Pathol. 1972 Jun;25(6):551–551. doi: 10.1136/jcp.25.6.551-a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mizunoe Y., Nakabeppu Y., Sekiguchi M., Kawabata S., Moriya T., Amako K. Cloning and sequence of the gene encoding the major structural component of mannose-resistant fimbriae of Serratia marcescens. J Bacteriol. 1988 Aug;170(8):3567–3574. doi: 10.1128/jb.170.8.3567-3574.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mobley H. L., Chippendale G. R. Hemagglutinin, urease, and hemolysin production by Proteus mirabilis from clinical sources. J Infect Dis. 1990 Mar;161(3):525–530. doi: 10.1093/infdis/161.3.525. [DOI] [PubMed] [Google Scholar]
  22. Mobley H. L., Chippendale G. R., Swihart K. G., Welch R. A. Cytotoxicity of the HpmA hemolysin and urease of Proteus mirabilis and Proteus vulgaris against cultured human renal proximal tubular epithelial cells. Infect Immun. 1991 Jun;59(6):2036–2042. doi: 10.1128/iai.59.6.2036-2042.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mobley H. L., Warren J. W. Urease-positive bacteriuria and obstruction of long-term urinary catheters. J Clin Microbiol. 1987 Nov;25(11):2216–2217. doi: 10.1128/jcm.25.11.2216-2217.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Moos M., Jr, Nguyen N. Y., Liu T. Y. Reproducible high yield sequencing of proteins electrophoretically separated and transferred to an inert support. J Biol Chem. 1988 May 5;263(13):6005–6008. [PubMed] [Google Scholar]
  25. Old D. C., Adegbola R. A. Haemagglutinins and fimbriae of Morganella, Proteus and Providencia. J Med Microbiol. 1982 Nov;15(4):551–564. doi: 10.1099/00222615-15-4-551. [DOI] [PubMed] [Google Scholar]
  26. Orndorff P. E., Falkow S. Nucleotide sequence of pilA, the gene encoding the structural component of type 1 pili in Escherichia coli. J Bacteriol. 1985 Apr;162(1):454–457. doi: 10.1128/jb.162.1.454-457.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Peerbooms P. G., Verweij A. M., MacLaren D. M. Investigation of the haemolytic activity of Proteus mirabilis strains. Antonie Van Leeuwenhoek. 1983 Apr;49(1):1–11. doi: 10.1007/BF00457874. [DOI] [PubMed] [Google Scholar]
  28. Peerbooms P. G., Verweij A. M., MacLaren D. M. Vero cell invasiveness of Proteus mirabilis. Infect Immun. 1984 Mar;43(3):1068–1071. doi: 10.1128/iai.43.3.1068-1071.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Poole K., Schiebel E., Braun V. Molecular characterization of the hemolysin determinant of Serratia marcescens. J Bacteriol. 1988 Jul;170(7):3177–3188. doi: 10.1128/jb.170.7.3177-3188.1988. [DOI] [PMC free article] [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. Sareneva T., Holthöfer H., Korhonen T. K. Tissue-binding affinity of Proteus mirabilis fimbriae in the human urinary tract. Infect Immun. 1990 Oct;58(10):3330–3336. doi: 10.1128/iai.58.10.3330-3336.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Setia U., Serventi I., Lorenz P. Bacteremia in a long-term care facility. Spectrum and mortality. Arch Intern Med. 1984 Aug;144(8):1633–1635. [PubMed] [Google Scholar]
  33. Silverblatt F. J. Host-parasite interaction in the rat renal pelvis: a possible role for pili in the pathogenesis of pyelonephritis. J Exp Med. 1974 Dec 1;140(6):1696–1711. doi: 10.1084/jem.140.6.1696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Silverblatt F. J., Ofek I. Influence of pili on the virulence of Proteus mirabilis in experimental hematogenous pyelonephritis. J Infect Dis. 1978 Nov;138(5):664–667. doi: 10.1093/infdis/138.5.664. [DOI] [PubMed] [Google Scholar]
  35. Swihart K. G., Welch R. A. Cytotoxic activity of the Proteus hemolysin HpmA. Infect Immun. 1990 Jun;58(6):1861–1869. doi: 10.1128/iai.58.6.1861-1869.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tacket C. O., Maneval D. R., Levine M. M. Purification, morphology, and genetics of a new fimbrial putative colonization factor of enterotoxigenic Escherichia coli O159:H4. Infect Immun. 1987 May;55(5):1063–1069. doi: 10.1128/iai.55.5.1063-1069.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Uphoff T. S., Welch R. A. Nucleotide sequencing of the Proteus mirabilis calcium-independent hemolysin genes (hpmA and hpmB) reveals sequence similarity with the Serratia marcescens hemolysin genes (shlA and shlB). J Bacteriol. 1990 Mar;172(3):1206–1216. doi: 10.1128/jb.172.3.1206-1216.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Voller A., Bidwell D., Huldt G., Engvall E. A microplate method of enzyme-linked immunosorbent assay and its application to malaria. Bull World Health Organ. 1974;51(2):209–211. [PMC free article] [PubMed] [Google Scholar]
  40. Warren J. W., Tenney J. H., Hoopes J. M., Muncie H. L., Anthony W. C. A prospective microbiologic study of bacteriuria in patients with chronic indwelling urethral catheters. J Infect Dis. 1982 Dec;146(6):719–723. doi: 10.1093/infdis/146.6.719. [DOI] [PubMed] [Google Scholar]
  41. Westerwoudt R. J. Improved fusion methods. IV. Technical aspects. J Immunol Methods. 1985 Mar 18;77(2):181–196. doi: 10.1016/0022-1759(85)90031-6. [DOI] [PubMed] [Google Scholar]
  42. Wray S. K., Hull S. I., Cook R. G., Barrish J., Hull R. A. Identification and characterization of a uroepithelial cell adhesin from a uropathogenic isolate of Proteus mirabilis. Infect Immun. 1986 Oct;54(1):43–49. doi: 10.1128/iai.54.1.43-49.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. van Die I., Bergmans H. Nucleotide sequence of the gene encoding the F72 fimbrial subunit of a uropathogenic Escherichia coli strain. Gene. 1984 Dec;32(1-2):83–90. doi: 10.1016/0378-1119(84)90035-0. [DOI] [PubMed] [Google Scholar]

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