Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1995 May;63(5):2082–2086. doi: 10.1128/iai.63.5.2082-2086.1995

Molecular cloning of Proteus mirabilis uroepithelial cell adherence (uca) genes.

S W Cook 1, N Mody 1, J Valle 1, R Hull 1
PMCID: PMC173269  PMID: 7729924

Abstract

Proteus mirabilis bacteria are a common cause of hospital-acquired urinary tract infection. In a previous study, we described a P. mirabilis fimbrial protein, UCA, that adhered to human uroepithelial cells. Genes sufficient for expression of UCA adherence were cloned into Escherichia coli K-12. E. coli bacteria that contained the uca recombinant plasmid adhered to human uroepithelial cells. In addition, the ucaA gene encoding the structural component of UCA pili was subcloned, and its DNA sequence was determined. Amino acid sequence homology (30 to 50%) was found between mature UcaA protein and pilins from pathogenic bacteria representing several genera, including E. coli F17, G, and type 1C pilins, Haemophilus M43 pilin, and a Bordetella pilin.

Full Text

The Full Text of this article is available as a PDF (251.7 KB).

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. Bahrani F. K., Cook S., Hull R. A., Massad G., Mobley H. L. Proteus mirabilis fimbriae: N-terminal amino acid sequence of a major fimbrial subunit and nucleotide sequences of the genes from two strains. Infect Immun. 1993 Mar;61(3):884–891. doi: 10.1128/iai.61.3.884-891.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bahrani F. K., Mobley H. L. Proteus mirabilis MR/P fimbriae: molecular cloning, expression, and nucleotide sequence of the major fimbrial subunit gene. J Bacteriol. 1993 Jan;175(2):457–464. doi: 10.1128/jb.175.2.457-464.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bruce A. W., Chan R. C., Pinkerton D., Morales A., Chadwick P. Adherence of gram-negative uropathogens to human uroepithelial cells. J Urol. 1983 Aug;130(2):293–298. doi: 10.1016/s0022-5347(17)51115-5. [DOI] [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. Gerlach G. F., Clegg S. Characterization of two genes encoding antigenically distinct type-1 fimbriae of Klebsiella pneumoniae. Gene. 1988 Apr 29;64(2):231–240. doi: 10.1016/0378-1119(88)90338-1. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Grosjean H., Fiers W. Preferential codon usage in prokaryotic genes: the optimal codon-anticodon interaction energy and the selective codon usage in efficiently expressed genes. Gene. 1982 Jun;18(3):199–209. doi: 10.1016/0378-1119(82)90157-3. [DOI] [PubMed] [Google Scholar]
  11. Hull R. A., Gill R. E., Hsu P., Minshew B. H., Falkow S. Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate. Infect Immun. 1981 Sep;33(3):933–938. doi: 10.1128/iai.33.3.933-938.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hull R. A., Hull S. I., Falkow S. Frequency of gene sequences necessary for pyelonephritis-associated pili expression among isolates of Enterobacteriaceae from human extraintestinal infections. Infect Immun. 1984 Mar;43(3):1064–1067. doi: 10.1128/iai.43.3.1064-1067.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johanson I., Lindstedt R., Svanborg C. Roles of the pap- and prs-encoded adhesins in Escherichia coli adherence to human uroepithelial cells. Infect Immun. 1992 Aug;60(8):3416–3422. doi: 10.1128/iai.60.8.3416-3422.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Johnson D. E., Russell R. G., Lockatell C. V., Zulty J. C., Warren J. W., Mobley H. L. Contribution of Proteus mirabilis urease to persistence, urolithiasis, and acute pyelonephritis in a mouse model of ascending urinary tract infection. Infect Immun. 1993 Jul;61(7):2748–2754. doi: 10.1128/iai.61.7.2748-2754.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jones B. D., Mobley H. L. Proteus mirabilis urease: nucleotide sequence determination and comparison with jack bean urease. J Bacteriol. 1989 Dec;171(12):6414–6422. doi: 10.1128/jb.171.12.6414-6422.1989. [DOI] [PMC free article] [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. Lintermans P., Pohl P., Deboeck F., Bertels A., Schlicker C., Vandekerckhove J., Van Damme J., Van Montagu M., De Greve H. Isolation and nucleotide sequence of the F17-A gene encoding the structural protein of the F17 fimbriae in bovine enterotoxigenic Escherichia coli. Infect Immun. 1988 Jun;56(6):1475–1484. doi: 10.1128/iai.56.6.1475-1484.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  19. Livey I., Duggleby C. J., Robinson A. Cloning and nucleotide sequence analysis of the serotype 2 fimbrial subunit gene of Bordetella pertussis. Mol Microbiol. 1987 Sep;1(2):203–209. doi: 10.1111/j.1365-2958.1987.tb00513.x. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Massad G., Bahrani F. K., Mobley H. L. Proteus mirabilis fimbriae: identification, isolation, and characterization of a new ambient-temperature fimbria. Infect Immun. 1994 May;62(5):1989–1994. doi: 10.1128/iai.62.5.1989-1994.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Nichols W. A., Clegg S., Brown M. R. Characterization of the type 1 fimbrial subunit gene (fimA) of Serratia marcescens. Mol Microbiol. 1990 Dec;4(12):2119–2126. doi: 10.1111/j.1365-2958.1990.tb00573.x. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Parsons C. L., Stauffer C., Mulholland S. G., Griffith D. P. Effect of ammonium on bacterial adherence to bladder transitional epithelium. J Urol. 1984 Aug;132(2):365–366. doi: 10.1016/s0022-5347(17)49628-5. [DOI] [PubMed] [Google Scholar]
  26. Pazin G. J., Braude A. I. Immobilizing antibodies in urine. II. Prevention of ascending spread of Proteus mirabilis. Invest Urol. 1974 Sep;12(2):129–133. [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. Rhen M., Klemm P., Korhonen T. K. Identification of two new hemagglutinins of Escherichia coli, N-acetyl-D-glucosamine-specific fimbriae and a blood group M-specific agglutinin, by cloning the corresponding genes in Escherichia coli K-12. J Bacteriol. 1986 Dec;168(3):1234–1242. doi: 10.1128/jb.168.3.1234-1242.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Senior B. W. The special affinity of particular types of Proteus mirabilis for the urinary tract. J Med Microbiol. 1979 Feb;12(1):1–8. doi: 10.1099/00222615-12-1-1. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. 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]
  35. Winkler H. H., Wood D. O. Codon usage in selected AT-rich bacteria. Biochimie. 1988 Aug;70(8):977–986. doi: 10.1016/0300-9084(88)90262-3. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. van Die I., van Geffen B., Hoekstra W., Bergmans H. Type 1C fimbriae of a uropathogenic Escherichia coli strain: cloning and characterization of the genes involved in the expression of the 1C antigen and nucleotide sequence of the subunit gene. Gene. 1985;34(2-3):187–196. doi: 10.1016/0378-1119(85)90127-1. [DOI] [PubMed] [Google Scholar]
  38. 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]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES