Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Sep;176(17):5385–5392. doi: 10.1128/jb.176.17.5385-5392.1994

Regulation of rns, a positive regulatory factor for pili of enterotoxigenic Escherichia coli.

B Froehlich 1, L Husmann 1, J Caron 1, J R Scott 1
PMCID: PMC196725  PMID: 7915269

Abstract

Attachment of enterotoxigenic Escherichia coli to the human gut is considered an important early step in infection that leads to diarrhea. This attachment is mediated by pili, which belong to a limited number of serologically distinguishable types. Many of these pili require the product of rns, or a closely related gene, for their expression. We have located the major promoter for rns and found that although its sequence diverges significantly from a sigma-70 promoter consensus sequence, it is very strong. Transcription of rns is negatively regulated both at a region upstream of this promoter and at a region internal to the rns open reading frame. In addition, rns positively regulates its own transcription, probably by counteracting these two negative effects.

Full text

PDF
5385

Images in this article

Selected References

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

  1. 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]
  2. Caron J., Maneval D. R., Kaper J. B., Scott J. R. Association of rns homologs with colonization factor antigens in clinical Escherichia coli isolates. Infect Immun. 1990 Oct;58(10):3442–3444. doi: 10.1128/iai.58.10.3442-3444.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
  5. Collado-Vides J., Magasanik B., Gralla J. D. Control site location and transcriptional regulation in Escherichia coli. Microbiol Rev. 1991 Sep;55(3):371–394. doi: 10.1128/mr.55.3.371-394.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cornelis G., Sluiters C., de Rouvroit C. L., Michiels T. Homology between virF, the transcriptional activator of the Yersinia virulence regulon, and AraC, the Escherichia coli arabinose operon regulator. J Bacteriol. 1989 Jan;171(1):254–262. doi: 10.1128/jb.171.1.254-262.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crothers D. M., Haran T. E., Nadeau J. G. Intrinsically bent DNA. J Biol Chem. 1990 May 5;265(13):7093–7096. [PubMed] [Google Scholar]
  8. Evans D. G., Evans D. J., Jr New surface-associated heat-labile colonization factor antigen (CFA/II) produced by enterotoxigenic Escherichia coli of serogroups O6 and O8. Infect Immun. 1978 Aug;21(2):638–647. doi: 10.1128/iai.21.2.638-647.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  11. Froehlich B. J., Scott J. R. A single-copy promoter-cloning vector for use in Escherichia coli. Gene. 1991 Dec 1;108(1):99–101. doi: 10.1016/0378-1119(91)90492-t. [DOI] [PubMed] [Google Scholar]
  12. Göransson M., Sondén B., Nilsson P., Dagberg B., Forsman K., Emanuelsson K., Uhlin B. E. Transcriptional silencing and thermoregulation of gene expression in Escherichia coli. Nature. 1990 Apr 12;344(6267):682–685. doi: 10.1038/344682a0. [DOI] [PubMed] [Google Scholar]
  13. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. He B., Zalkin H. Repression of Escherichia coli purB is by a transcriptional roadblock mechanism. J Bacteriol. 1992 Nov;174(22):7121–7127. doi: 10.1128/jb.174.22.7121-7127.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jordi B. J., Dagberg B., de Haan L. A., Hamers A. M., van der Zeijst B. A., Gaastra W., Uhlin B. E. The positive regulator CfaD overcomes the repression mediated by histone-like protein H-NS (H1) in the CFA/I fimbrial operon of Escherichia coli. EMBO J. 1992 Jul;11(7):2627–2632. doi: 10.1002/j.1460-2075.1992.tb05328.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Levine M. M. Escherichia coli that cause diarrhea: enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent. J Infect Dis. 1987 Mar;155(3):377–389. doi: 10.1093/infdis/155.3.377. [DOI] [PubMed] [Google Scholar]
  18. Levine M. M., Ristaino P., Marley G., Smyth C., Knutton S., Boedeker E., Black R., Young C., Clements M. L., Cheney C. Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans. Infect Immun. 1984 May;44(2):409–420. doi: 10.1128/iai.44.2.409-420.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Linn T., St Pierre R. Improved vector system for constructing transcriptional fusions that ensures independent translation of lacZ. J Bacteriol. 1990 Feb;172(2):1077–1084. doi: 10.1128/jb.172.2.1077-1084.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Macdonald P. M., Kutter E., Mosig G. Regulation of a bacteriophage T4 late gene, soc, which maps in an early region. Genetics. 1984 Jan;106(1):17–27. doi: 10.1093/genetics/106.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. McClure W. R. Mechanism and control of transcription initiation in prokaryotes. Annu Rev Biochem. 1985;54:171–204. doi: 10.1146/annurev.bi.54.070185.001131. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  24. Morschhäuser J., Uhlin B. E., Hacker J. Transcriptional analysis and regulation of the sfa determinant coding for S fimbriae of pathogenic Escherichia coli strains. Mol Gen Genet. 1993 Apr;238(1-2):97–105. doi: 10.1007/BF00279536. [DOI] [PubMed] [Google Scholar]
  25. Ogierman M. A., Manning P. A. Homology of TcpN, a putative regulatory protein of Vibrio cholerae, to the AraC family of transcriptional activators. Gene. 1992 Jul 1;116(1):93–97. doi: 10.1016/0378-1119(92)90634-2. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Plaskon R. R., Wartell R. M. Sequence distributions associated with DNA curvature are found upstream of strong E. coli promoters. Nucleic Acids Res. 1987 Jan 26;15(2):785–796. doi: 10.1093/nar/15.2.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ross W., Gosink K. K., Salomon J., Igarashi K., Zou C., Ishihama A., Severinov K., Gourse R. L. A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. Science. 1993 Nov 26;262(5138):1407–1413. doi: 10.1126/science.8248780. [DOI] [PubMed] [Google Scholar]
  29. Sakai T., Sasakawa C., Makino S., Yoshikawa M. DNA sequence and product analysis of the virF locus responsible for congo red binding and cell invasion in Shigella flexneri 2a. Infect Immun. 1986 Nov;54(2):395–402. doi: 10.1128/iai.54.2.395-402.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Scott J. R. A turbid plaque-forming mutant of phage P1 that cannot lysogenize Escherichia coli. Virology. 1974 Dec;62(2):344–349. doi: 10.1016/0042-6822(74)90397-3. [DOI] [PubMed] [Google Scholar]
  33. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Smyth C. J. Two mannose-resistant haemagglutinins on enterotoxigenic Escherichia coli of serotype O6:K15:H16 or H-isolated from travellers' and infantile diarrhoea. J Gen Microbiol. 1982 Sep;128(9):2081–2096. doi: 10.1099/00221287-128-9-2081. [DOI] [PubMed] [Google Scholar]
  35. Takeshita S., Sato M., Toba M., Masahashi W., Hashimoto-Gotoh T. High-copy-number and low-copy-number plasmid vectors for lacZ alpha-complementation and chloramphenicol- or kanamycin-resistance selection. Gene. 1987;61(1):63–74. doi: 10.1016/0378-1119(87)90365-9. [DOI] [PubMed] [Google Scholar]
  36. Thomas L. V., McConnell M. M., Rowe B., Field A. M. The possession of three novel coli surface antigens by enterotoxigenic Escherichia coli strains positive for the putative colonization factor PCF8775. J Gen Microbiol. 1985 Sep;131(9):2319–2326. doi: 10.1099/00221287-131-9-2319. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Willshaw G. A., Smith H. R., McConnell M. M., Rowe B. Cloning of regulator genes controlling fimbrial production by enterotoxigenic Escherichia coli. FEMS Microbiol Lett. 1991 Aug 1;66(2):125–129. doi: 10.1016/0378-1097(91)90320-a. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. de Haan L. A., Willshaw G. A., van der Zeijst B. A., Gaastra W. The nucleotide sequence of a regulatory gene present on a plasmid in an enterotoxigenic Escherichia coli strain of serotype O167:H5. FEMS Microbiol Lett. 1991 Oct 15;67(3):341–346. doi: 10.1111/j.1574-6968.1991.tb04487.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES