Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Jul;178(14):4273–4280. doi: 10.1128/jb.178.14.4273-4280.1996

Identification of the promoter region for the colanic acid polysaccharide biosynthetic genes in Escherichia coli K-12.

V Stout 1
PMCID: PMC178186  PMID: 8763957

Abstract

The colanic acid polysaccharide capsule biosynthetic genes (cps genes) are primarily clustered at one site located at about 45 min on the Escherichia coli chromosome. The network of proteins involved in regulating the expression of these genes includes the two positive regulators RcsA and RcsB. This work describes the site of action of these two activator proteins and the promoter of the cps genes. It is likely that the cps genes are arranged in a single long operon that is at least 13.5 kb. The promoter region was identified with fusions to lacZ that resulted in regulated expression by the Rcs network of regulatory proteins, and the start site of transcription was identified by primer extension. The operator region was cloned from Kohara phage to multicopy plasmids and identified by titrating RcsA or RcsB. Sequence analysis of the promoter and operator region revealed homology to the JUMPstart element found in the untranslated region of many exopolysaccharide biosynthetic operons. In addition, the deduced amino acid sequence of the amino terminus of the first open reading frame of the cps operon was found to be homologous to proteins encoded by the exopolysaccharide biosynthetic operons of Klebsiella pneumoniae and Erwinia amylovora.

Full Text

The Full Text of this article is available as a PDF (367.8 KB).

Selected References

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

  1. Allen P., Hart C. A., Saunders J. R. Isolation from Klebsiella and characterization of two rcs genes that activate colanic acid capsular biosynthesis in Escherichia coli. J Gen Microbiol. 1987 Feb;133(2):331–340. doi: 10.1099/00221287-133-2-331. [DOI] [PubMed] [Google Scholar]
  2. Arakawa Y., Wacharotayankun R., Nagatsuka T., Ito H., Kato N., Ohta M. Genomic organization of the Klebsiella pneumoniae cps region responsible for serotype K2 capsular polysaccharide synthesis in the virulent strain Chedid. J Bacteriol. 1995 Apr;177(7):1788–1796. doi: 10.1128/jb.177.7.1788-1796.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bernhard F., Poetter K., Geider K., Coplin D. L. The rcsA gene from Erwinia amylovora: identification, nucleotide sequence, and regulation of exopolysaccharide biosynthesis. Mol Plant Microbe Interact. 1990 Nov-Dec;3(6):429–437. doi: 10.1094/mpmi-3-429. [DOI] [PubMed] [Google Scholar]
  4. Brill J. A., Quinlan-Walshe C., Gottesman S. Fine-structure mapping and identification of two regulators of capsule synthesis in Escherichia coli K-12. J Bacteriol. 1988 Jun;170(6):2599–2611. doi: 10.1128/jb.170.6.2599-2611.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bugert P., Geider K. Molecular analysis of the ams operon required for exopolysaccharide synthesis of Erwinia amylovora. Mol Microbiol. 1995 Mar;15(5):917–933. doi: 10.1111/j.1365-2958.1995.tb02361.x. [DOI] [PubMed] [Google Scholar]
  6. Coleman M., Pearce R., Hitchin E., Busfield F., Mansfield J. W., Roberts I. S. Molecular cloning, expression and nucleotide sequence of the rcsA gene of Erwinia amylovora, encoding a positive regulator of capsule expression: evidence for a family of related capsule activator proteins. J Gen Microbiol. 1990 Sep;136(9):1799–1806. doi: 10.1099/00221287-136-9-1799. [DOI] [PubMed] [Google Scholar]
  7. Frosch M., Edwards U., Bousset K., Krausse B., Weisgerber C. Evidence for a common molecular origin of the capsule gene loci in gram-negative bacteria expressing group II capsular polysaccharides. Mol Microbiol. 1991 May;5(5):1251–1263. doi: 10.1111/j.1365-2958.1991.tb01899.x. [DOI] [PubMed] [Google Scholar]
  8. GOEBEL W. F. Colanic acid. Proc Natl Acad Sci U S A. 1963 Apr;49:464–471. doi: 10.1073/pnas.49.4.464. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gervais F. G., Phoenix P., Drapeau G. R. The rcsB gene, a positive regulator of colanic acid biosynthesis in Escherichia coli, is also an activator of ftsZ expression. J Bacteriol. 1992 Jun;174(12):3964–3971. doi: 10.1128/jb.174.12.3964-3971.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gottesman S., Stout V. Regulation of capsular polysaccharide synthesis in Escherichia coli K12. Mol Microbiol. 1991 Jul;5(7):1599–1606. doi: 10.1111/j.1365-2958.1991.tb01906.x. [DOI] [PubMed] [Google Scholar]
  11. Gottesman S., Trisler P., Torres-Cabassa A. Regulation of capsular polysaccharide synthesis in Escherichia coli K-12: characterization of three regulatory genes. J Bacteriol. 1985 Jun;162(3):1111–1119. doi: 10.1128/jb.162.3.1111-1119.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Hinton D. M. Transcript analyses of the uvsX-40-41 region of bacteriophage T4. Changes in the RNA as infection proceeds. J Biol Chem. 1989 Aug 25;264(24):14432–14439. [PubMed] [Google Scholar]
  14. Hobbs M., Reeves P. R. The JUMPstart sequence: a 39 bp element common to several polysaccharide gene clusters. Mol Microbiol. 1994 Jun;12(5):855–856. doi: 10.1111/j.1365-2958.1994.tb01071.x. [DOI] [PubMed] [Google Scholar]
  15. Houng H. S., Noon K. F., Ou J. T., Baron L. S. Expression of Vi antigen in Escherichia coli K-12: characterization of ViaB from Citrobacter freundii and identity of ViaA with RcsB. J Bacteriol. 1992 Sep;174(18):5910–5915. doi: 10.1128/jb.174.18.5910-5915.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jann K., Jann B. Polysaccharide antigens of Escherichia coli. Rev Infect Dis. 1987 Sep-Oct;9 (Suppl 5):S517–S526. doi: 10.1093/clinids/9.supplement_5.s517. [DOI] [PubMed] [Google Scholar]
  17. Jayaratne P., Keenleyside W. J., MacLachlan P. R., Dodgson C., Whitfield C. Characterization of rcsB and rcsC from Escherichia coli O9:K30:H12 and examination of the role of the rcs regulatory system in expression of group I capsular polysaccharides. J Bacteriol. 1993 Sep;175(17):5384–5394. doi: 10.1128/jb.175.17.5384-5394.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Keenleyside W. J., Jayaratne P., MacLachlan P. R., Whitfield C. The rcsA gene of Escherichia coli O9:K30:H12 is involved in the expression of the serotype-specific group I K (capsular) antigen. J Bacteriol. 1992 Jan;174(1):8–16. doi: 10.1128/jb.174.1.8-16.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  20. Kroll J. S., Loynds B., Brophy L. N., Moxon E. R. The bex locus in encapsulated Haemophilus influenzae: a chromosomal region involved in capsule polysaccharide export. Mol Microbiol. 1990 Nov;4(11):1853–1862. doi: 10.1111/j.1365-2958.1990.tb02034.x. [DOI] [PubMed] [Google Scholar]
  21. Maurizi M. R., Trisler P., Gottesman S. Insertional mutagenesis of the lon gene in Escherichia coli: lon is dispensable. J Bacteriol. 1985 Dec;164(3):1124–1135. doi: 10.1128/jb.164.3.1124-1135.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McCallum K. L., Whitfield C. The rcsA gene of Klebsiella pneumoniae O1:K20 is involved in expression of the serotype-specific K (capsular) antigen. Infect Immun. 1991 Feb;59(2):494–502. doi: 10.1128/iai.59.2.494-502.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Poetter K., Coplin D. L. Structural and functional analysis of the rcsA gene from Erwinia stewartii. Mol Gen Genet. 1991 Sep;229(1):155–160. doi: 10.1007/BF00264225. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
  26. Stout V., Gottesman S. RcsB and RcsC: a two-component regulator of capsule synthesis in Escherichia coli. J Bacteriol. 1990 Feb;172(2):659–669. doi: 10.1128/jb.172.2.659-669.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stout V., Torres-Cabassa A., Maurizi M. R., Gutnick D., Gottesman S. RcsA, an unstable positive regulator of capsular polysaccharide synthesis. J Bacteriol. 1991 Mar;173(5):1738–1747. doi: 10.1128/jb.173.5.1738-1747.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Torres-Cabassa A. S., Gottesman S. Capsule synthesis in Escherichia coli K-12 is regulated by proteolysis. J Bacteriol. 1987 Mar;169(3):981–989. doi: 10.1128/jb.169.3.981-989.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Trisler P., Gottesman S. lon transcriptional regulation of genes necessary for capsular polysaccharide synthesis in Escherichia coli K-12. J Bacteriol. 1984 Oct;160(1):184–191. doi: 10.1128/jb.160.1.184-191.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zinkewich-Péotti K., Fraser J. M. New locus for exopolysaccharide overproduction in Escherichia coli K-12. J Bacteriol. 1988 Mar;170(3):1405–1407. doi: 10.1128/jb.170.3.1405-1407.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES