Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Feb;65(2):507–513. doi: 10.1128/iai.65.2.507-513.1997

A new putative fimbrial colonization factor, CS19, of human enterotoxigenic Escherichia coli.

H M Grewal 1, H Valvatne 1, M K Bhan 1, L van Dijk 1, W Gaastra 1, H Sommerfelt 1
PMCID: PMC176088  PMID: 9009305

Abstract

A gene probe derived from the colonization factor antigen I (CFA/I) operon cross-hybridized at very low stringency to plasmid DNA from coli surface antigen 17 (CS17)-producing enterotoxigenic Escherichia coli (ETEC) and from the ETEC strain F595C, which was negative for previously described CFAs, CSs, and putative colonization factors (PCFs). A 16-kDa protein was identified in sodium dodecyl sulfate-polyacrylamide gel electrophoresis of heat extracts prepared after growth of strain F595C at 37 degrees C on CFA agar containing bile salts. Transmission electron microscopy revealed bile salt- and temperature-dependent expression of fimbriae with a diameter of 7 nm. After transformation with a recombinant plasmid harboring the cfaR gene, which encodes a positive regulator of several CFAs, PCFs, and CSs, the 16-kDa protein was hyperexpressed. Polyclonal antibodies raised against this protein bound to the fimbriae and inhibited the adhesion of F595C bacteria to tissue-cultured Caco-2 cells. Nucleotide sequence determination of the gene encoding the 16-kDa fimbrial subunit revealed a high degree of amino acid sequence homology to the CFA/I, CS1, CS2, CS4, CS14, and CS17 polypeptides. The term CS19 is proposed for the new fimbria.

Full Text

The Full Text of this article is available as a PDF (844.4 KB).

Selected References

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

  1. Aubel D., Darfeuille-Michaud A., Joly B. New adhesive factor (antigen 8786) on a human enterotoxigenic Escherichia coli O117:H4 strain isolated in Africa. Infect Immun. 1991 Apr;59(4):1290–1299. doi: 10.1128/iai.59.4.1290-1299.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bergman T., Jörnvall H. Electroblotting of individual polypeptides from SDS/polyacrylamide gels for direct sequence analysis. Eur J Biochem. 1987 Nov 16;169(1):9–12. doi: 10.1111/j.1432-1033.1987.tb13573.x. [DOI] [PubMed] [Google Scholar]
  3. Binsztein N., Jouve M. J., Viboud G. I., López Moral L., Rivas M., Orskov I., Ahrén C., Svennerholm A. M. Colonization factors of enterotoxigenic Escherichia coli isolated from children with diarrhea in Argentina. J Clin Microbiol. 1991 Sep;29(9):1893–1898. doi: 10.1128/jcm.29.9.1893-1898.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Black R. E. Epidemiology of diarrhoeal disease: implications for control by vaccines. Vaccine. 1993;11(2):100–106. doi: 10.1016/0264-410x(93)90002-f. [DOI] [PubMed] [Google Scholar]
  5. Caron J., Coffield L. M., Scott J. R. A plasmid-encoded regulatory gene, rns, required for expression of the CS1 and CS2 adhesins of enterotoxigenic Escherichia coli. Proc Natl Acad Sci U S A. 1989 Feb;86(3):963–967. doi: 10.1073/pnas.86.3.963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Caron J., Scott J. R. A rns-like regulatory gene for colonization factor antigen I (CFA/I) that controls expression of CFA/I pilin. Infect Immun. 1990 Apr;58(4):874–878. doi: 10.1128/iai.58.4.874-878.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clemens J. D., Sack D. A., Harris J. R., Chakraborty J., Neogy P. K., Stanton B., Huda N., Khan M. U., Kay B. A., Khan M. R. Cross-protection by B subunit-whole cell cholera vaccine against diarrhea associated with heat-labile toxin-producing enterotoxigenic Escherichia coli: results of a large-scale field trial. J Infect Dis. 1988 Aug;158(2):372–377. doi: 10.1093/infdis/158.2.372. [DOI] [PubMed] [Google Scholar]
  8. Darfeuille-Michaud A., Aubel D., Chauviere G., Rich C., Bourges M., Servin A., Joly B. Adhesion of enterotoxigenic Escherichia coli to the human colon carcinoma cell line Caco-2 in culture. Infect Immun. 1990 Apr;58(4):893–902. doi: 10.1128/iai.58.4.893-902.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Darfeuille-Michaud A., Forestier C., Joly B., Cluzel R. Identification of a nonfimbrial adhesive factor of an enterotoxigenic Escherichia coli strain. Infect Immun. 1986 May;52(2):468–475. doi: 10.1128/iai.52.2.468-475.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Elsinghorst E. A., Kopecko D. J. Molecular cloning of epithelial cell invasion determinants from enterotoxigenic Escherichia coli. Infect Immun. 1992 Jun;60(6):2409–2417. doi: 10.1128/iai.60.6.2409-2417.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Evans D. G., Evans D. J., Jr, Tjoa W. Hemagglutination of human group A erythrocytes by enterotoxigenic Escherichia coli isolated from adults with diarrhea: correlation with colonization factor. Infect Immun. 1977 Nov;18(2):330–337. doi: 10.1128/iai.18.2.330-337.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Evans D. G., Silver R. P., Evans D. J., Jr, Chase D. G., Gorbach S. L. Plasmid-controlled colonization factor associated with virulence in Esherichia coli enterotoxigenic for humans. Infect Immun. 1975 Sep;12(3):656–667. doi: 10.1128/iai.12.3.656-667.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Froehlich B. J., Karakashian A., Sakellaris H., Scott J. R. Genes for CS2 pili of enterotoxigenic Escherichia coli and their interchangeability with those for CS1 pili. Infect Immun. 1995 Dec;63(12):4849–4856. doi: 10.1128/iai.63.12.4849-4856.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gaastra W., de Graaf F. K. Host-specific fimbrial adhesins of noninvasive enterotoxigenic Escherichia coli strains. Microbiol Rev. 1982 Jun;46(2):129–161. doi: 10.1128/mr.46.2.129-161.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grewal H. M., Gaastra W., Svennerholm A. M., Röli J., Sommerfelt H. Induction of colonization factor antigen I (CFA/I) and coli surface antigen 4 (CS4) of enterotoxigenic Escherichia coli: relevance for vaccine production. Vaccine. 1993;11(2):221–226. doi: 10.1016/0264-410x(93)90021-o. [DOI] [PubMed] [Google Scholar]
  16. Grewal H. M., Helander A., Svennerholm A. M., Bhan M. K., Gaastra W., Sommerfelt H. Genotypic and phenotypic identification of coli surface antigen 6-positive enterotoxigenic Escherichia coli. J Clin Microbiol. 1994 May;32(5):1295–1301. doi: 10.1128/jcm.32.5.1295-1301.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Grewal H. M., Sommerfelt H., Gaastra W., Svennerholm A. M., Bhan M. K., Hamers A. M., Kumar R., Wiklund G., Bjorvatn B. Detection of colonization factor antigen I-positive enterotoxigenic Escherichia coli with a cloned polynucleotide probe. J Clin Microbiol. 1990 Oct;28(10):2264–2268. doi: 10.1128/jcm.28.10.2264-2268.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hamers A. M., Pel H. J., Willshaw G. A., Kusters J. G., van der Zeijst B. A., Gaastra W. The nucleotide sequence of the first two genes of the CFA/I fimbrial operon of human enterotoxigenic Escherichia coli. Microb Pathog. 1989 Apr;6(4):297–309. doi: 10.1016/0882-4010(89)90103-4. [DOI] [PubMed] [Google Scholar]
  19. Heuzenroeder M. W., Elliot T. R., Thomas C. J., Halter R., Manning P. A. A new fimbrial type (PCFO9) on enterotoxigenic Escherichia coli 09:H- LT+ isolated from a case of infant diarrhea in central Australia. FEMS Microbiol Lett. 1990 Jan 1;54(1-3):55–60. doi: 10.1016/0378-1097(90)90258-r. [DOI] [PubMed] [Google Scholar]
  20. Hibberd M. L., McConnell M. M., Field A. M., Rowe B. The fimbriae of human enterotoxigenic Escherichia coli strain 334 are related to CS5 fimbriae. J Gen Microbiol. 1990 Dec;136(12):2449–2456. doi: 10.1099/00221287-136-12-2449. [DOI] [PubMed] [Google Scholar]
  21. Hibberd M. L., McConnell M. M., Willshaw G. A., Smith H. R., Rowe B. Positive regulation of colonization factor antigen I (CFA/I) production by enterotoxigenic Escherichia coli producing the colonization factors CS5, CS6, CS7, CS17, PCFO9, PCFO159:H4 and PCFO166. J Gen Microbiol. 1991 Aug;137(8):1963–1970. doi: 10.1099/00221287-137-8-1963. [DOI] [PubMed] [Google Scholar]
  22. Honda T., Arita M., Miwatani T. Characterization of new hydrophobic pili of human enterotoxigenic Escherichia coli: a possible new colonization factor. Infect Immun. 1984 Mar;43(3):959–965. doi: 10.1128/iai.43.3.959-965.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Karjalainen T. K., Evans D. G., So M., Lee C. H. Molecular cloning and nucleotide sequence of the colonization factor antigen I gene of Escherichia coli. Infect Immun. 1989 Apr;57(4):1126–1130. doi: 10.1128/iai.57.4.1126-1130.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Knutton S., Lloyd D. R., McNeish A. S. Identification of a new fimbrial structure in enterotoxigenic Escherichia coli (ETEC) serotype O148:H28 which adheres to human intestinal mucosa: a potentially new human ETEC colonization factor. Infect Immun. 1987 Jan;55(1):86–92. doi: 10.1128/iai.55.1.86-92.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Lopez-Vidal Y., Svennerholm A. M. Monoclonal antibodies against the different subcomponents of colonization factor antigen II of enterotoxigenic Escherichia coli. J Clin Microbiol. 1990 Sep;28(9):1906–1912. doi: 10.1128/jcm.28.9.1906-1912.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Marron M. B., Smyth C. J. Molecular analysis of the cso operon of enterotoxigenic Escherichia coli reveals that CsoA is the adhesin of CS1 fimbriae and that the accessory genes are interchangeable with those of the cfa operon. Microbiology. 1995 Nov;141(Pt 11):2849–2859. doi: 10.1099/13500872-141-11-2849. [DOI] [PubMed] [Google Scholar]
  28. McConnell M. M., Chart H., Field A. M., Hibberd M., Rowe B. Characterization of a putative colonization factor (PCFO166) of enterotoxigenic Escherichia coli of serogroup O166. J Gen Microbiol. 1989 May;135(5):1135–1144. doi: 10.1099/00221287-135-5-1135. [DOI] [PubMed] [Google Scholar]
  29. McConnell M. M., Hibberd M., Field A. M., Chart H., Rowe B. Characterization of a new putative colonization factor (CS17) from a human enterotoxigenic Escherichia coli of serotype O114:H21 which produces only heat-labile enterotoxin. J Infect Dis. 1990 Feb;161(2):343–347. doi: 10.1093/infdis/161.2.343. [DOI] [PubMed] [Google Scholar]
  30. McConnell M. M., Thomas L. V., Willshaw G. A., Smith H. R., Rowe B. Genetic control and properties of coli surface antigens of colonization factor antigen IV (PCF8775) of enterotoxigenic Escherichia coli. Infect Immun. 1988 Aug;56(8):1974–1980. doi: 10.1128/iai.56.8.1974-1980.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mooi F. R., de Graaf F. K. Molecular biology of fimbriae of enterotoxigenic Escherichia coli. Curr Top Microbiol Immunol. 1985;118:119–138. doi: 10.1007/978-3-642-70586-1_7. [DOI] [PubMed] [Google Scholar]
  32. Ochman H., Gerber A. S., Hartl D. L. Genetic applications of an inverse polymerase chain reaction. Genetics. 1988 Nov;120(3):621–623. doi: 10.1093/genetics/120.3.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Perez-Casal J., Swartley J. S., Scott J. R. Gene encoding the major subunit of CS1 pili of human enterotoxigenic Escherichia coli. Infect Immun. 1990 Nov;58(11):3594–3600. doi: 10.1128/iai.58.11.3594-3600.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Savelkoul P. H., Willshaw G. A., McConnell M. M., Smith H. R., Hamers A. M., van der Zeijst B. A., Gaastra W. Expression of CFA/I fimbriae is positively regulated. Microb Pathog. 1990 Feb;8(2):91–99. doi: 10.1016/0882-4010(90)90073-y. [DOI] [PubMed] [Google Scholar]
  35. Smith H. R., Willshaw G. A., Rowe B. Mapping of a plasmid, coding for colonization, factor antigen I and heat-stable enterotoxin production, isolated from an enterotoxigenic strain of Escherichia coli. J Bacteriol. 1982 Jan;149(1):264–275. doi: 10.1128/jb.149.1.264-275.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sommerfelt H., Grewal H. M., Bhan M. K. Simplified and accurate nonradioactive polynucleotide gene probe assay for identification of enterotoxigenic Escherichia coli. J Clin Microbiol. 1990 Jan;28(1):49–54. doi: 10.1128/jcm.28.1.49-54.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sommerfelt H., Grewal H. M., Gaastra W., Bhan M. K., Svennerholm A. M., Kalland K. H., Asphaug V., Aasland R., Bjorvatn B. Presence of cfaD-homologous sequences and expression of coli surface antigen 4 on enterotoxigenic Escherichia coli; relevance for diagnostic procedures. Microb Pathog. 1991 Oct;11(4):297–304. doi: 10.1016/0882-4010(91)90034-8. [DOI] [PubMed] [Google Scholar]
  38. Sommerfelt H., Grewal H. M., Gaastra W., Svennerholm A. M., Bhan M. K. Use of nonradioactive DNA hybridization for identification of enterotoxigenic Escherichia coli harboring genes for colonization factor antigen I, coli surface antigen 4, or putative colonization factor O166. J Clin Microbiol. 1992 Jul;30(7):1823–1828. doi: 10.1128/jcm.30.7.1823-1828.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Sommerfelt H., Grewal H. M., Svennerholm A. M., Gaastra W., Flood P. R., Viboud G., Bhan M. K. Genetic relationship of putative colonization factor O166 to colonization factor antigen I and coli surface antigen 4 of enterotoxigenic Escherichia coli. Infect Immun. 1992 Sep;60(9):3799–3806. doi: 10.1128/iai.60.9.3799-3806.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sommerfelt H., Steinsland H., Grewal H. M., Viboud G. I., Bhandari N., Gaastra W., Svennerholm A. M., Bhan M. K. Colonization factors of enterotoxigenic Escherichia coli isolated from children in north India. J Infect Dis. 1996 Oct;174(4):768–776. doi: 10.1093/infdis/174.4.768. [DOI] [PubMed] [Google Scholar]
  41. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  42. Svennerholm A. M., Holmgren J., Sack D. A. Development of oral vaccines against enterotoxinogenic Escherichia coli diarrhoea. Vaccine. 1989 Jun;7(3):196–198. doi: 10.1016/0264-410x(89)90228-4. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Valvatne H., Sommerfelt H., Gaastra W., Bhan M. K., Grewal H. M. Identification and characterization of CS20, a new putative colonization factor of enterotoxigenic Escherichia coli. Infect Immun. 1996 Jul;64(7):2635–2642. doi: 10.1128/iai.64.7.2635-2642.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Viboud G. I., Binsztein N., Svennerholm A. M. A new fimbrial putative colonization factor, PCFO20, in human enterotoxigenic Escherichia coli. Infect Immun. 1993 Dec;61(12):5190–5197. doi: 10.1128/iai.61.12.5190-5197.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Viboud G. I., Binsztein N., Svennerholm A. M. Characterization of monoclonal antibodies against putative colonization factors of enterotoxigenic Escherichia coli and their use in an epidemiological study. J Clin Microbiol. 1993 Mar;31(3):558–564. doi: 10.1128/jcm.31.3.558-564.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Viboud G. I., McConnell M. M., Helander A., Svennerholm A. M. Binding of enterotoxigenic Escherichia coli expressing different colonization factors to tissue-cultured Caco-2 cells and to isolated human enterocytes. Microb Pathog. 1996 Aug;21(2):139–147. doi: 10.1006/mpat.1996.0049. [DOI] [PubMed] [Google Scholar]
  48. Willshaw G. A., McConnell M. M., Smith H. R., Rowe B. Structural and regulatory genes for coli surface associated antigen 4 (CS4) are encoded by separate plasmids in enterotoxigenic Escherichia coli strains of serotype 0.25.H42. FEMS Microbiol Lett. 1990 Mar 15;56(3):255–260. [PubMed] [Google Scholar]
  49. Wolf M. K., Andrews G. P., Tall B. D., McConnell M. M., Levine M. M., Boedeker E. C. Characterization of CS4 and CS6 antigenic components of PCF8775, a putative colonization factor complex from enterotoxigenic Escherichia coli E8775. Infect Immun. 1989 Jan;57(1):164–173. doi: 10.1128/iai.57.1.164-173.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES