Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Feb;174(3):1063–1067. doi: 10.1128/jb.174.3.1063-1067.1992

Involvement of rcsB in Klebsiella K2 capsule synthesis in Escherichia coli K-12.

R Wacharotayankun 1, Y Arakawa 1, M Ohta 1, T Hasegawa 1, M Mori 1, T Horii 1, N Kato 1
PMCID: PMC206199  PMID: 1732199

Abstract

Escherichia coli K-12 harboring a part of the structural genes for the Klebsiella K2 capsular polysaccharide (cpsK*) expresses a large amount of K2 capsular polysaccharide as a thick capsule in the presence of plasmids carrying rmpA and rcsB. We have previously shown that expression of the Klebsiella K2 capsule in E. coli HB101 harboring cpsK* depends on the presence of rmpA, a regulatory gene from a large plasmid of Klebsiella pneumoniae Chedid (O1:K2). E. coli K-12 JM109, however, produces only a small amount of K2 capsular polysaccharide, even in the presence of plasmids carrying rmpA as well as the cpsK* structural genes. Introduction of the rcsB gene, a positive regulator of colanic acid capsule synthesis in E. coli K-12 which was cloned from HB101 on a plasmid, into JM109 cells carrying cpsK* and rmpA, results in the expression of a thick K2 capsule. By Northern (RNA) hybridization analysis, rcsB has been found to enhance transcription of a long strand of mRNA (longer than 14 kb) from cpsK*. These E. coli transformants which produce a thick K2 capsule also express colanic acid production at high levels. Therefore, rcsB can act as a positive regulator of Klebsiella K2 capsule production and two capsular polysaccharides can be expressed in E. coli simultaneously. With a somewhat different strain background, we have found that both of the colanic acid regulators, rcsA and rcsB, contribute to the basal level of Klebsiella K2 capsule expression but that the presence of multicopy rcsB in either an rcsB or an rcsA mutant of E. coli is sufficient to increase the expression of K2 capsular polysaccharide. These results suggest further parallels between the regulation of colanic acid synthesis in E. coli and the regulation of Klebsiella K2 capsule synthesis.

Full text

PDF

Images in this article

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., Ohta M., Kido N., Mori M., Ito H., Komatsu T., Fujii Y., Kato N. Chromosomal beta-lactamase of Klebsiella oxytoca, a new class A enzyme that hydrolyzes broad-spectrum beta-lactam antibiotics. Antimicrob Agents Chemother. 1989 Jan;33(1):63–70. doi: 10.1128/aac.33.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Arakawa Y., Ohta M., Wacharotayankun R., Mori M., Kido N., Ito H., Komatsu T., Sugiyama T., Kato N. Biosynthesis of Klebsiella K2 capsular polysaccharide in Escherichia coli HB101 requires the functions of rmpA and the chromosomal cps gene cluster of the virulent strain Klebsiella pneumoniae Chedid (O1:K2). Infect Immun. 1991 Jun;59(6):2043–2050. doi: 10.1128/iai.59.6.2043-2050.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Arakawa Y., Wacharotayankun R., Ohta M., Shoji K., Watahiki M., Horii T., Kato N. Construction of a novel suicide vector: selection for Escherichia coli HB101 recombinants carrying the DNA insert. Gene. 1991 Jul 31;104(1):81–84. doi: 10.1016/0378-1119(91)90468-q. [DOI] [PubMed] [Google Scholar]
  5. BATSHON B. A., BAER H., SHAFFER M. F. Immunologic paralysis produced in mice by Klebsiella pneumoniae type 2 polysaccharide. J Immunol. 1963 Jan;90:121–126. [PubMed] [Google Scholar]
  6. Bolivar F., Backman K. Plasmids of Escherichia coli as cloning vectors. Methods Enzymol. 1979;68:245–267. doi: 10.1016/0076-6879(79)68018-7. [DOI] [PubMed] [Google Scholar]
  7. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Cryz S. J., Jr, Mortimer P. M., Mansfield V., Germanier R. Seroepidemiology of Klebsiella bacteremic isolates and implications for vaccine development. J Clin Microbiol. 1986 Apr;23(4):687–690. doi: 10.1128/jcm.23.4.687-690.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Edmondson A. S., Cooke E. M. The production of antisera to the Klebsiella capsular antigens. J Appl Bacteriol. 1979 Jun;46(3):579–584. doi: 10.1111/j.1365-2672.1979.tb00858.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. Kiseleva B. S., Krasnogolovets V. N. Rol' Klebsiella pneumoniae v étiologii bakterial'nogo sepsisa. Zh Mikrobiol Epidemiol Immunobiol. 1983 Feb;(2):20–25. [PubMed] [Google Scholar]
  13. Kornblum J. S., Projan S. J., Moghazeh S. L., Novick R. P. A rapid method to quantitate non-labeled RNA species in bacterial cells. Gene. 1988;63(1):75–85. doi: 10.1016/0378-1119(88)90547-1. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Mizuta K., Ohta M., Mori M., Hasegawa T., Nakashima I., Kato N. Virulence for mice of Klebsiella strains belonging to the O1 group: relationship to their capsular (K) types. Infect Immun. 1983 Apr;40(1):56–61. doi: 10.1128/iai.40.1.56-61.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mori M., Ohta M., Agata N., Kido N., Arakawa Y., Ito H., Komatsu T., Kato N. Identification of species and capsular types of Klebsiella clinical isolates, with special reference to Klebsiella planticola. Microbiol Immunol. 1989;33(11):887–895. doi: 10.1111/j.1348-0421.1989.tb00976.x. [DOI] [PubMed] [Google Scholar]
  17. Nassif X., Honoré N., Vasselon T., Cole S. T., Sansonetti P. J. Positive control of colanic acid synthesis in Escherichia coli by rmpA and rmpB, two virulence-plasmid genes of Klebsiella pneumoniae. Mol Microbiol. 1989 Oct;3(10):1349–1359. doi: 10.1111/j.1365-2958.1989.tb00116.x. [DOI] [PubMed] [Google Scholar]
  18. Simoons-Smit A. M., Verweij-van Vught A. M., MacLaren D. M. The role of K antigens as virulence factors in Klebsiella. J Med Microbiol. 1986 Mar;21(2):133–137. doi: 10.1099/00222615-21-2-133. [DOI] [PubMed] [Google Scholar]
  19. Stauffer G. V., Plamann M. D., Stauffer L. T. Construction and expression of hybrid plasmids containing the Escherichia coli glyA genes. Gene. 1981 Jun-Jul;14(1-2):63–72. doi: 10.1016/0378-1119(81)90148-7. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Sugiyama T., Kido N., Arakawa Y., Mori M., Naito S., Ohta M., Kato N. Rapid small-scale preparation method of cell surface polysaccharides. Microbiol Immunol. 1990;34(7):635–641. doi: 10.1111/j.1348-0421.1990.tb01039.x. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. 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]
  26. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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

RESOURCES