Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1996 Jul;64(7):2635–2642. doi: 10.1128/iai.64.7.2635-2642.1996

Identification and characterization of CS20, a new putative colonization factor of enterotoxigenic Escherichia coli.

H Valvatne 1, H Sommerfelt 1, W Gaastra 1, M K Bhan 1, H M Grewal 1
PMCID: PMC174120  PMID: 8698489

Abstract

An enterotoxigenic Escherichia coli (ETEC) strain producing a previously undescribed putative colonization factor was isolated from a child with diarrhea in India. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of bacterial heat extracts revealed a polypeptide band of 20.8 kDa when the bacteria were grown at 37 degrees C which was absent after growth at 22 degrees C. A specific rabbit antiserum raised against the purified 20.8-kDa protein bound specifically to the fimbriae, as shown by immunoelectron microscopy, and inhibited bacterial adhesion to tissue-cultured Caco-2 cells. Transformation with a recombinant plasmid harboring the cfaD gene, which encodes a positive regulator for several ETEC fimbriae, induced hyperexpression of the 20.8-kDa fimbrial subunit and a substantial increase in the proportion of bacterial cells that were fimbriated. The N-terminal amino acid sequence of the polypeptide showed 65 and 60% identity to the PCFO20 and 987P fimbriae of human and porcine ETEC, respectively. We propose the term CS20 for this new putative colonization factor of human ETEC.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Ahrén C. M., Svennerholm A. M. Experimental enterotoxin-induced Escherichia coli diarrhea and protection induced by previous infection with bacteria of the same adhesin or enterotoxin type. Infect Immun. 1985 Oct;50(1):255–261. doi: 10.1128/iai.50.1.255-261.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  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. Blanco J., Blanco M., Garabal J. I., González E. A. Enterotoxins, colonization factors and serotypes of enterotoxigenic Escherichia coli from humans and animals. Microbiologia. 1991 Sep;7(2):57–73. [PubMed] [Google Scholar]
  6. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  7. Cravioto A., Reyes R. E., Trujillo F., Uribe F., Navarro A., De La Roca J. M., Hernández J. M., Pérez G., Vázquez V. Risk of diarrhea during the first year of life associated with initial and subsequent colonization by specific enteropathogens. Am J Epidemiol. 1990 May;131(5):886–904. doi: 10.1093/oxfordjournals.aje.a115579. [DOI] [PubMed] [Google Scholar]
  8. Cravioto A., Scotland S. M., Rowe B. Hemagglutination activity and colonization factor antigens I and II in enterotoxigenic and non-enterotoxigenic strains of Escherichia coli isolated from humans. Infect Immun. 1982 Apr;36(1):189–197. doi: 10.1128/iai.36.1.189-197.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  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. Girón J. A., Levine M. M., Kaper J. B. Longus: a long pilus ultrastructure produced by human enterotoxigenic Escherichia coli. Mol Microbiol. 1994 Apr;12(1):71–82. doi: 10.1111/j.1365-2958.1994.tb00996.x. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. 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]
  17. 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]
  18. 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]
  19. 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]
  20. 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]
  21. 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]
  22. Isaacson R. E., Nagy B., Moon H. W. Colonization of porcine small intestine by Escherichia coli: colonization and adhesion factors of pig enteropathogens that lack K88. J Infect Dis. 1977 Apr;135(4):531–539. doi: 10.1093/infdis/135.4.531. [DOI] [PubMed] [Google Scholar]
  23. Klaasen P., Woodward M. J., van Zijderveld F. G., de Graaf F. K. The 987P gene cluster in enterotoxigenic Escherichia coli contains an STpa transposon that activates 987P expression. Infect Immun. 1990 Mar;58(3):801–807. doi: 10.1128/iai.58.3.801-807.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Klaasen P., de Graaf F. K. Characterization of FapR, a positive regulator of expression of the 987P operon in enterotoxigenic Escherichia coli. Mol Microbiol. 1990 Oct;4(10):1779–1783. doi: 10.1111/j.1365-2958.1990.tb00556.x. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. 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]
  28. McConnell M. M., Hibberd M. L., Penny M. E., Scotland S. M., Cheasty T., Rowe B. Surveys of human enterotoxigenic Escherichia coli from three different geographical areas for possible colonization factors. Epidemiol Infect. 1991 Jun;106(3):477–484. doi: 10.1017/s0950268800067522. [DOI] [PMC free article] [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. Moon H. W., Bunn T. O. Vaccines for preventing enterotoxigenic Escherichia coli infections in farm animals. Vaccine. 1993;11(2):213–200. doi: 10.1016/0264-410X(93)90020-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nagy B., Moon H. W., Isaacson R. E. Colonization of porcine small intestine by Escherichia coli: ileal colonization and adhesion by pig enteropathogens that lack K88 antigen and by some acapsular mutants. Infect Immun. 1976 Apr;13(4):1214–1220. doi: 10.1128/iai.13.4.1214-1220.1976. [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. 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]
  36. 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]
  37. 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]
  38. Svennerholm A. M., McConnell M. M., Wiklund G. Roles of different putative colonization factor antigens in colonization of human enterotoxigenic Escherichia coli in rabbits. Microb Pathog. 1992 Nov;13(5):381–389. doi: 10.1016/0882-4010(92)90081-x. [DOI] [PubMed] [Google Scholar]
  39. Svennerholm A. M., Vidal Y. L., Holmgren J., McConnell M. M., Rowe B. Role of PCF8775 antigen and its coli surface subcomponents for colonization, disease, and protective immunogenicity of enterotoxigenic Escherichia coli in rabbits. Infect Immun. 1988 Feb;56(2):523–528. doi: 10.1128/iai.56.2.523-528.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. 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]
  41. 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]
  42. 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]
  43. Yamamoto T., Gojobori T., Yokota T. Evolutionary origin of pathogenic determinants in enterotoxigenic Escherichia coli and Vibrio cholerae O1. J Bacteriol. 1987 Mar;169(3):1352–1357. doi: 10.1128/jb.169.3.1352-1357.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Zabarovsky E. R., Winberg G. High efficiency electroporation of ligated DNA into bacteria. Nucleic Acids Res. 1990 Oct 11;18(19):5912–5912. doi: 10.1093/nar/18.19.5912. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES