Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Jul;65(7):2685–2692. doi: 10.1128/iai.65.7.2685-2692.1997

Evolutionary relationships among pathogenic and nonpathogenic Escherichia coli strains inferred from multilocus enzyme electrophoresis and mdh sequence studies.

G M Pupo 1, D K Karaolis 1, R Lan 1, P R Reeves 1
PMCID: PMC175379  PMID: 9199437

Abstract

Within the species Escherichia coli, there are commensal strains and a variety of pathogenic strains, including enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and urinary tract infection (UTI) strains. The pathogenic strains are identified by serotype and by possession of specific virulence determinants (toxins and adhesions, etc.) encoded by either monocistronic genes, plasmids, or pathogenicity islands. Although there are studies on the relationships between selected pathogenic strains, the relatedness among the majority of the pathogenic forms to each other, to commensal E. coli, and to the genus Shigella (which has often been suggested to be part of E. coli) has not been determined. We used multilocus enzyme electrophoresis (MLEE) at 10 enzyme loci and the sequence of the mdh housekeeping gene to study the genetic relationships of pathogenic E. coli strains (including Shigella clones), namely, 5 EPEC strains (serotypes O111 and O55), 3 EHEC strains (serotype O157), 6 ETEC strains (serotypes O78, O159, and O148), 5 EIEC strains (serotypes O124, O28, and O112), and 13 Shigella strains representing clones Flexneri, Dysenteriae, Boydii, and Sonnei, to commensal E. coli strains. Both the MLEE and mdh sequence trees reveal that EPEC, EHEC, ETEC, EIEC, and UTI strains are distributed among the ECOR set groups, with no overall clustering of EPEC, ETEC, EIEC, or UTI strains. The genus Shigella is shown to comprise a group of closely related pathogenic E. coli strains. Six pathogenic strains, i.e., M502 (EIEC; O112ac:NM), M503 (EPEC; O111:H12), M526 (ETEC; O159:H4), M522 (EPEC; O111ac:H12), M524 (ETEC; O78:H11), and M506 (ETEC; O78:H11), were found to have mdh sequences identical to those of five ECOR group A strains (ECOR5, ECOR10, ECOR14, ECOR6, and K-12). All 11 strains are closely related by MLEE. The results indicate that pathogenic strains of E. coli do not have a single evolutionary origin within E. coli but have arisen many times. The results also suggest the possibility that any E. coli strain acquiring the appropriate virulence factors may give rise to a pathogenic form.

Full Text

The Full Text of this article is available as a PDF (269.5 KB).

Selected References

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

  1. Bastin D. A., Romana L. K., Reeves P. R. Molecular cloning and expression in Escherichia coli K-12 of the rfb gene cluster determining the O antigen of an E. coli O111 strain. Mol Microbiol. 1991 Sep;5(9):2223–2231. doi: 10.1111/j.1365-2958.1991.tb02152.x. [DOI] [PubMed] [Google Scholar]
  2. Boyd E. F., Nelson K., Wang F. S., Whittam T. S., Selander R. K. Molecular genetic basis of allelic polymorphism in malate dehydrogenase (mdh) in natural populations of Escherichia coli and Salmonella enterica. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1280–1284. doi: 10.1073/pnas.91.4.1280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brenner D. J., Fanning G. R., Skerman F. J., Falkow S. Polynucleotide sequence divergence among strains of Escherichia coli and closely related organisms. J Bacteriol. 1972 Mar;109(3):953–965. doi: 10.1128/jb.109.3.953-965.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brenner D. J., Fanning G. R., Steigerwalt A. G., Orskov I., Orskov F. Polynucleotide sequence relatedness among three groups of pathogenic Escherichia coli strains. Infect Immun. 1972 Sep;6(3):308–315. doi: 10.1128/iai.6.3.308-315.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cordovéz A., Prado V., Maggi L., Cordero J., Martinez J., Misraji A., Rios R., Soza G., Ojeda A., Levine M. M. Enterohemorrhagic Escherichia coli associated with hemolytic-uremic syndrome in Chilean children. J Clin Microbiol. 1992 Aug;30(8):2153–2157. doi: 10.1128/jcm.30.8.2153-2157.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Donnenberg M. S., Kaper J. B. Enteropathogenic Escherichia coli. Infect Immun. 1992 Oct;60(10):3953–3961. doi: 10.1128/iai.60.10.3953-3961.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Donnenberg M. S., Tzipori S., McKee M. L., O'Brien A. D., Alroy J., Kaper J. B. The role of the eae gene of enterohemorrhagic Escherichia coli in intimate attachment in vitro and in a porcine model. J Clin Invest. 1993 Sep;92(3):1418–1424. doi: 10.1172/JCI116718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. EWING W. H. Serological relationships between shigella and coliform cultures. J Bacteriol. 1953 Sep;66(3):333–340. doi: 10.1128/jb.66.3.333-340.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elliott E., Li Z., Bell C., Stiel D., Buret A., Wallace J., Brzuszczak I., O'Loughlin E. Modulation of host response to Escherichia coli o157:H7 infection by anti-CD18 antibody in rabbits. Gastroenterology. 1994 Jun;106(6):1554–1561. doi: 10.1016/0016-5085(94)90410-3. [DOI] [PubMed] [Google Scholar]
  10. Fasano A., Kay B. A., Russell R. G., Maneval D. R., Jr, Levin M. M. Enterotoxin and cytotoxin production by enteroinvasive Escherichia coli. Infect Immun. 1990 Nov;58(11):3717–3723. doi: 10.1128/iai.58.11.3717-3723.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goullet P. Esterase electrophoretic pattern relatedness between Shigella species and Escherichia coli. J Gen Microbiol. 1980 Apr;117(2):493–500. doi: 10.1099/00221287-117-2-493. [DOI] [PubMed] [Google Scholar]
  12. Hartl D. L., Dykhuizen D. E. The population genetics of Escherichia coli. Annu Rev Genet. 1984;18:31–68. doi: 10.1146/annurev.ge.18.120184.000335. [DOI] [PubMed] [Google Scholar]
  13. Herzer P. J., Inouye S., Inouye M., Whittam T. S. Phylogenetic distribution of branched RNA-linked multicopy single-stranded DNA among natural isolates of Escherichia coli. J Bacteriol. 1990 Nov;172(11):6175–6181. doi: 10.1128/jb.172.11.6175-6181.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Karaolis D. K., Lan R., Reeves P. R. Sequence variation in Shigella sonnei (Sonnei), a pathogenic clone of Escherichia coli, over four continents and 41 years. J Clin Microbiol. 1994 Mar;32(3):796–802. doi: 10.1128/jcm.32.3.796-802.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Katzenellenbogen E., Romanowska E. Structural studies on Shigella flexneri serotype 6 core region. Eur J Biochem. 1980 Dec;113(1):205–211. doi: 10.1111/j.1432-1033.1980.tb06157.x. [DOI] [PubMed] [Google Scholar]
  16. Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol. 1980 Dec;16(2):111–120. doi: 10.1007/BF01731581. [DOI] [PubMed] [Google Scholar]
  17. Knutton S., Baldwin T., Williams P. H., McNeish A. S. Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli. Infect Immun. 1989 Apr;57(4):1290–1298. doi: 10.1128/iai.57.4.1290-1298.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Konowalchuk J., Speirs J. I., Stavric S. Vero response to a cytotoxin of Escherichia coli. Infect Immun. 1977 Dec;18(3):775–779. doi: 10.1128/iai.18.3.775-779.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lawrence J. G., Ochman H., Hartl D. L. Molecular and evolutionary relationships among enteric bacteria. J Gen Microbiol. 1991 Aug;137(8):1911–1921. doi: 10.1099/00221287-137-8-1911. [DOI] [PubMed] [Google Scholar]
  20. Levine M. M., Edelman R. Enteropathogenic Escherichia coli of classic serotypes associated with infant diarrhea: epidemiology and pathogenesis. Epidemiol Rev. 1984;6:31–51. doi: 10.1093/oxfordjournals.epirev.a036274. [DOI] [PubMed] [Google Scholar]
  21. Levine M. M., Kaper J. B., Black R. E., Clements M. L. New knowledge on pathogenesis of bacterial enteric infections as applied to vaccine development. Microbiol Rev. 1983 Dec;47(4):510–550. doi: 10.1128/mr.47.4.510-550.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lund B., Lindberg F., Marklund B. I., Normark S. The PapG protein is the alpha-D-galactopyranosyl-(1----4)-beta-D-galactopyranose-binding adhesin of uropathogenic Escherichia coli. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5898–5902. doi: 10.1073/pnas.84.16.5898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lund B., Marklund B. I., Strömberg N., Lindberg F., Karlsson K. A., Normark S. Uropathogenic Escherichia coli can express serologically identical pili of different receptor binding specificities. Mol Microbiol. 1988 Mar;2(2):255–263. doi: 10.1111/j.1365-2958.1988.tb00027.x. [DOI] [PubMed] [Google Scholar]
  24. McDaniel T. K., Jarvis K. G., Donnenberg M. S., Kaper J. B. A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1664–1668. doi: 10.1073/pnas.92.5.1664. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Milkman R. Electrophoretic variation in Escherichia coli from natural sources. Science. 1973 Dec 7;182(4116):1024–1026. doi: 10.1126/science.182.4116.1024. [DOI] [PubMed] [Google Scholar]
  26. Moon H. W., Whipp S. C., Argenzio R. A., Levine M. M., Giannella R. A. Attaching and effacing activities of rabbit and human enteropathogenic Escherichia coli in pig and rabbit intestines. Infect Immun. 1983 Sep;41(3):1340–1351. doi: 10.1128/iai.41.3.1340-1351.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. NETER E., WESTPHAL O., LUDERITZ O., GINO R. M., GORZYNSKI E. A. Demonstration of antibodies against enteropathogenic Escherichia coli in sera of children of various ages. Pediatrics. 1955 Dec;16(6):801–808. [PubMed] [Google Scholar]
  28. Nasoff M. S., Baker H. V., 2nd, Wolf R. E., Jr DNA sequence of the Escherichia coli gene, gnd, for 6-phosphogluconate dehydrogenase. Gene. 1984 Mar;27(3):253–264. doi: 10.1016/0378-1119(84)90070-2. [DOI] [PubMed] [Google Scholar]
  29. Nelson K., Selander R. K. Evolutionary genetics of the proline permease gene (putP) and the control region of the proline utilization operon in populations of Salmonella and Escherichia coli. J Bacteriol. 1992 Nov;174(21):6886–6895. doi: 10.1128/jb.174.21.6886-6895.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nelson K., Whittam T. S., Selander R. K. Nucleotide polymorphism and evolution in the glyceraldehyde-3-phosphate dehydrogenase gene (gapA) in natural populations of Salmonella and Escherichia coli. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6667–6671. doi: 10.1073/pnas.88.15.6667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. O'Brien A. D., Melton A. R., Schmitt C. K., McKee M. L., Batts M. L., Griffin D. E. Profile of Escherichia coli O157:H7 pathogen responsible for hamburger-borne outbreak of hemorrhagic colitis and hemolytic uremic syndrome in Washington. J Clin Microbiol. 1993 Oct;31(10):2799–2801. doi: 10.1128/jcm.31.10.2799-2801.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ochman H., Selander R. K. Evidence for clonal population structure in Escherichia coli. Proc Natl Acad Sci U S A. 1984 Jan;81(1):198–201. doi: 10.1073/pnas.81.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ochman H., Whittam T. S., Caugant D. A., Selander R. K. Enzyme polymorphism and genetic population structure in Escherichia coli and Shigella. J Gen Microbiol. 1983 Sep;129(9):2715–2726. doi: 10.1099/00221287-129-9-2715. [DOI] [PubMed] [Google Scholar]
  34. Orskov F., Whittam T. S., Cravioto A., Orskov I. Clonal relationships among classic enteropathogenic Escherichia coli (EPEC) belong to different O groups. J Infect Dis. 1990 Jul;162(1):76–81. doi: 10.1093/infdis/162.1.76. [DOI] [PubMed] [Google Scholar]
  35. Ostroff S. M., Tarr P. I., Neill M. A., Lewis J. H., Hargrett-Bean N., Kobayashi J. M. Toxin genotypes and plasmid profiles as determinants of systemic sequelae in Escherichia coli O157:H7 infections. J Infect Dis. 1989 Dec;160(6):994–998. doi: 10.1093/infdis/160.6.994. [DOI] [PubMed] [Google Scholar]
  36. Polotsky Y. E., Dragunskaya E. M., Seliverstova V. G., Avdeeva T. A., Chakhutinskaya M. G., Kétyi I., Vertényl A., Ralovich B., Emödy L., Málovics I. Pathogenic effect of enterotoxigenic Escherichia coli and Escherichia coli causing infantile diarrhoea. Acta Microbiol Acad Sci Hung. 1977;24(3):221–236. [PubMed] [Google Scholar]
  37. Reeves P. R., Farnell L., Lan R. MULTICOMP: a program for preparing sequence data for phylogenetic analysis. Comput Appl Biosci. 1994 Jun;10(3):281–284. doi: 10.1093/bioinformatics/10.3.281. [DOI] [PubMed] [Google Scholar]
  38. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  39. Sansonetti P. J., Kopecko D. J., Formal S. B. Involvement of a plasmid in the invasive ability of Shigella flexneri. Infect Immun. 1982 Mar;35(3):852–860. doi: 10.1128/iai.35.3.852-860.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sansonetti P. J., Kopecko D. J., Formal S. B. Shigella sonnei plasmids: evidence that a large plasmid is necessary for virulence. Infect Immun. 1981 Oct;34(1):75–83. doi: 10.1128/iai.34.1.75-83.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sansonetti P. J., d'Hauteville H., Ecobichon C., Pourcel C. Molecular comparison of virulence plasmids in Shigella and enteroinvasive Escherichia coli. Ann Microbiol (Paris) 1983 May-Jun;134A(3):295–318. [PubMed] [Google Scholar]
  42. Selander R. K., Korhonen T. K., Väisänen-Rhen V., Williams P. H., Pattison P. E., Caugant D. A. Genetic relationships and clonal structure of strains of Escherichia coli causing neonatal septicemia and meningitis. Infect Immun. 1986 Apr;52(1):213–222. doi: 10.1128/iai.52.1.213-222.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sherman P., Soni R., Karmali M. Attaching and effacing adherence of Vero cytotoxin-producing Escherichia coli to rabbit intestinal epithelium in vivo. Infect Immun. 1988 Apr;56(4):756–761. doi: 10.1128/iai.56.4.756-761.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Stevenson G., Neal B., Liu D., Hobbs M., Packer N. H., Batley M., Redmond J. W., Lindquist L., Reeves P. Structure of the O antigen of Escherichia coli K-12 and the sequence of its rfb gene cluster. J Bacteriol. 1994 Jul;176(13):4144–4156. doi: 10.1128/jb.176.13.4144-4156.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. TAYLOR J., POWELL B. W., WRIGHT J. Infantile diarrhoea and vomiting; a clinical and bacteriological investigation. Br Med J. 1949 Jul 16;2(4619):117–125. doi: 10.1136/bmj.2.4619.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tzipori S., Wachsmuth I. K., Chapman C., Birden R., Brittingham J., Jackson C., Hogg J. The pathogenesis of hemorrhagic colitis caused by Escherichia coli O157:H7 in gnotobiotic piglets. J Infect Dis. 1986 Oct;154(4):712–716. doi: 10.1093/infdis/154.4.712. [DOI] [PubMed] [Google Scholar]
  47. Welch R. A., Hull R., Falkow S. Molecular cloning and physical characterization of a chromosomal hemolysin from Escherichia coli. Infect Immun. 1983 Oct;42(1):178–186. doi: 10.1128/iai.42.1.178-186.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Whittam T. S., Ochman H., Selander R. K. Multilocus genetic structure in natural populations of Escherichia coli. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1751–1755. doi: 10.1073/pnas.80.6.1751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Whittam T. S., Wachsmuth I. K., Wilson R. A. Genetic evidence of clonal descent of Escherichia coli O157:H7 associated with hemorrhagic colitis and hemolytic uremic syndrome. J Infect Dis. 1988 Jun;157(6):1124–1133. doi: 10.1093/infdis/157.6.1124. [DOI] [PubMed] [Google Scholar]
  50. Whittam T. S., Wilson R. A. Genetic relationships among pathogenic Escherichia coli of serogroup O157. Infect Immun. 1988 Sep;56(9):2467–2473. doi: 10.1128/iai.56.9.2467-2473.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Whittam T. S., Wilson R. A. Genetic relationships among pathogenic strains of avian Escherichia coli. Infect Immun. 1988 Sep;56(9):2458–2466. doi: 10.1128/iai.56.9.2458-2466.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Whittam T. S., Wolfe M. L., Wachsmuth I. K., Orskov F., Orskov I., Wilson R. A. Clonal relationships among Escherichia coli strains that cause hemorrhagic colitis and infantile diarrhea. Infect Immun. 1993 May;61(5):1619–1629. doi: 10.1128/iai.61.5.1619-1629.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES