Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1988 Aug;170(8):3567–3574. doi: 10.1128/jb.170.8.3567-3574.1988

Cloning and sequence of the gene encoding the major structural component of mannose-resistant fimbriae of Serratia marcescens.

Y Mizunoe 1, Y Nakabeppu 1, M Sekiguchi 1, S Kawabata 1, T Moriya 1, K Amako 1
PMCID: PMC211330  PMID: 2900238

Abstract

Serratia marcescens US46, a human urinary tract isolate, exhibits mannose-resistant hemagglutination and agglutinates yeast cells, thereby indicating that it has two types of adhesins. We constructed a cosmid library for the DNA of this organism and isolated DNA clones carrying genes for mannose-sensitive (MS) and mannose-resistant (MR) fimbriae. On introduction of the cloned genes into Escherichia coli K-12, MS and MR fimbriae were formed. These fimbriae were functionally and morphologically indistinguishable from those of S. marcescens. Subcloning of these gene clusters revealed that the genes encoding MS fimbriae reside on a 9-kilobase (kb) DNA fragment, while those encoding MR fimbriae are present on a 12-kb fragment. Transposon insertion and maxicell analyses revealed that formation of MR fimbriae is controlled by several genes which reside on the 9-kb fragment. The nucleotide sequence of smfA, the gene encoding the major structural component of MR fimbriae, revealed that this gene encodes a 174-amino-acid polypeptide with a typical procaryotic signal peptide. The primary structure of the smfA product showed significant homology with the primary structure of the E. coli fimbrial subunit.

Full text

PDF
3567

Images in this article

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. New fimbrial hemagglutinin in Serratia species. Infect Immun. 1982 Oct;38(1):306–315. doi: 10.1128/iai.38.1.306-315.1982. [DOI] [PMC free article] [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. Allen S. D., Conger K. B. Serratia marcescens infection of the urinary tract: a nosocomial infection. J Urol. 1969 Apr;101(4):621–623. doi: 10.1016/s0022-5347(17)62391-7. [DOI] [PubMed] [Google Scholar]
  4. BERNS K. I., THOMAS C. A., Jr ISOLATION OF HIGH MOLECULAR WEIGHT DNA FROM HEMOPHILUS INFLUENZAE. J Mol Biol. 1965 Mar;11:476–490. doi: 10.1016/s0022-2836(65)80004-3. [DOI] [PubMed] [Google Scholar]
  5. Barnes W. M., Bevan M., Son P. H. Kilo-sequencing: creation of an ordered nest of asymmetric deletions across a large target sequence carried on phage M13. Methods Enzymol. 1983;101:98–122. doi: 10.1016/0076-6879(83)01008-3. [DOI] [PubMed] [Google Scholar]
  6. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  7. Båga M., Göransson M., Normark S., Uhlin B. E. Transcriptional activation of a pap pilus virulence operon from uropathogenic Escherichia coli. EMBO J. 1985 Dec 30;4(13B):3887–3893. doi: 10.1002/j.1460-2075.1985.tb04162.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Clegg S. Cloning of genes determining the production of mannose-resistant fimbriae in a uropathogenic strain of Escherichia coli belonging to serogroup O6. Infect Immun. 1982 Nov;38(2):739–744. doi: 10.1128/iai.38.2.739-744.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Clegg S., Hull S., Hull R., Pruckler J. Construction and comparison of recombinant plasmids encoding type 1 fimbriae of members of the family Enterobacteriaceae. Infect Immun. 1985 May;48(2):275–279. doi: 10.1128/iai.48.2.275-279.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Eden C. S., Eriksson B., Hanson L. A. Adhesion of Escherichia coli to human uroepithelial cells in vitro. Infect Immun. 1977 Dec;18(3):767–774. doi: 10.1128/iai.18.3.767-774.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Evans D. J., Jr, Evans D. G., DuPont H. L. Hemagglutination patterns of enterotoxigenic and enteropathogenic Escherichia coli determined with human, bovine, chicken, and guinea pig erythrocytes in the presence and absence of mannose. Infect Immun. 1979 Feb;23(2):336–346. doi: 10.1128/iai.23.2.336-346.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fader R. C., Davis C. P. Effect of piliation on Klebsiella pneumoniae infection in rat bladders. Infect Immun. 1980 Nov;30(2):554–561. doi: 10.1128/iai.30.2.554-561.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Farmer J. J., 3rd, Davis B. R., Hickman F. W., Presley D. B., Bodey G. P., Negut M., Bobo R. A. Detection of Serratia outbreaks in hospital. Lancet. 1976 Aug 28;2(7983):455–459. doi: 10.1016/s0140-6736(76)92539-3. [DOI] [PubMed] [Google Scholar]
  16. GALE D., SONNENWIRTH A. C. Frequent human isolation of Serratia marcescens. Bacteriological and pathogenicity studies of twelve strains of S. marcescens recovered from nine patients during a six-month period. Arch Intern Med. 1962 Apr;109:414–421. doi: 10.1001/archinte.1962.03620160040006. [DOI] [PubMed] [Google Scholar]
  17. Hacker J., Schmidt G., Hughes C., Knapp S., Marget M., Goebel W. Cloning and characterization of genes involved in production of mannose-resistant, neuraminidase-susceptible (X) fimbriae from a uropathogenic O6:K15:H31 Escherichia coli strain. Infect Immun. 1985 Feb;47(2):434–440. doi: 10.1128/iai.47.2.434-440.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  19. Hattori M., Sakaki Y. Dideoxy sequencing method using denatured plasmid templates. Anal Biochem. 1986 Feb 1;152(2):232–238. doi: 10.1016/0003-2697(86)90403-3. [DOI] [PubMed] [Google Scholar]
  20. Hermodson M. A., Chen K. C., Buchanan T. M. Neisseria pili proteins: amino-terminal amino acid sequences and identification of an unusual amino acid. Biochemistry. 1978 Feb 7;17(3):442–445. doi: 10.1021/bi00596a010. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Hohn B., Murray K. Packaging recombinant DNA molecules into bacteriophage particles in vitro. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3259–3263. doi: 10.1073/pnas.74.8.3259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Jingushi S., Mitsuyama M., Moriya T., Amako K. Antigenic analysis of Serratia marcescens fimbriae with monoclonal antibodies. Infect Immun. 1987 Jul;55(7):1600–1606. doi: 10.1128/iai.55.7.1600-1606.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kohno K., Yamamoto T., Kuroiwa A., Amako K. Purification and characterization of Serratia marcescens US5 pili. Infect Immun. 1984 Nov;46(2):295–300. doi: 10.1128/iai.46.2.295-300.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. Maki D. G., Hennekens C. G., Phillips C. W., Shaw W. V., Bennett J. V. Nosocomial urinary tract infection with Serratia marcescens: an epidemiologic study. J Infect Dis. 1973 Nov;128(5):579–587. doi: 10.1093/infdis/128.5.579. [DOI] [PubMed] [Google Scholar]
  30. Marrs C. F., Schoolnik G., Koomey J. M., Hardy J., Rothbard J., Falkow S. Cloning and sequencing of a Moraxella bovis pilin gene. J Bacteriol. 1985 Jul;163(1):132–139. doi: 10.1128/jb.163.1.132-139.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. McKern N. M., O'Donnell I. J., Inglis A. S., Stewart D. J., Clark B. L. Amino acid sequence of pilin from Bacteroides nodosus (strain 198), the causative organism of ovine footrot. FEBS Lett. 1983 Nov 28;164(1):149–153. doi: 10.1016/0014-5793(83)80039-8. [DOI] [PubMed] [Google Scholar]
  32. Meyerowitz E. M., Guild G. M., Prestidge L. S., Hogness D. S. A new high-capacity cosmid vector and its use. Gene. 1980 Nov;11(3-4):271–282. doi: 10.1016/0378-1119(80)90067-0. [DOI] [PubMed] [Google Scholar]
  33. Mooi F. R., Wouters C., Wijfjes A., de Graaf F. K. Construction and characterization of mutants impaired in the biosynthesis of the K88ab antigen. J Bacteriol. 1982 May;150(2):512–521. doi: 10.1128/jb.150.2.512-521.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Orndorff P. E., Falkow S. Organization and expression of genes responsible for type 1 piliation in Escherichia coli. J Bacteriol. 1984 Aug;159(2):736–744. doi: 10.1128/jb.159.2.736-744.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rhen M., van Die I., Rhen V., Bergmans H. Comparison of the nucleotide sequences of the genes encoding the KS71A and F7(1) fimbrial antigens of uropathogenic Escherichia coli. Eur J Biochem. 1985 Sep 16;151(3):573–577. doi: 10.1111/j.1432-1033.1985.tb09142.x. [DOI] [PubMed] [Google Scholar]
  37. Sakumi K., Nakabeppu Y., Yamamoto Y., Kawabata S., Iwanaga S., Sekiguchi M. Purification and structure of 3-methyladenine-DNA glycosylase I of Escherichia coli. J Biol Chem. 1986 Nov 25;261(33):15761–15766. [PubMed] [Google Scholar]
  38. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Yamamoto T., Ariyoshi A., Amako K. Fimbria-mediated adherence of Serratia marcescens strain US5 to human urinary bladder surface. Microbiol Immunol. 1985;29(7):677–681. doi: 10.1111/j.1348-0421.1985.tb00871.x. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. de Graaf F. K., Krenn B. E., Klaasen P. Organization and expression of genes involved in the biosynthesis of K99 fimbriae. Infect Immun. 1984 Feb;43(2):508–514. doi: 10.1128/iai.43.2.508-514.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. 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]
  45. van der Bosch J. F., Verboom-Sohmer U., Postma P., de Graaff J., MacLaren D. M. Mannose-sensitive and mannose-resistant adherence to human uroepithelial cells and urinary virulence of Escherichia coli. Infect Immun. 1980 Jul;29(1):226–233. doi: 10.1128/iai.29.1.226-233.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. von Heijne G. How signal sequences maintain cleavage specificity. J Mol Biol. 1984 Feb 25;173(2):243–251. doi: 10.1016/0022-2836(84)90192-x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES