Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1991 Feb;59(2):494–502. doi: 10.1128/iai.59.2.494-502.1991

The rcsA gene of Klebsiella pneumoniae O1:K20 is involved in expression of the serotype-specific K (capsular) antigen.

K L McCallum 1, C Whitfield 1
PMCID: PMC257777  PMID: 1987069

Abstract

In Klebsiella pneumoniae, the ability to synthesize large amounts of capsular polysaccharide is an important correlate of virulence. We report the cloning of rcsA from K. pneumoniae serotype O1:K20 and demonstrate that rcsA is involved in the expression of the K antigen capsule. We have determined the nucleotide sequence for the rcsA gene from K. pneumoniae K20 and shown it to be identical to the sequence reported previously for rcsA from strain K21 (Allen et al., J. Gen. Microbiol. 133:331-340, 1987). Southern hybridization results indicate that this gene is widely distributed among different Klebsiella K serotypes. When cloned into Escherichia coli K-12, the K. pneumoniae rcsA gene caused a mucoid phenotype, resulting from the activation of colanic acid synthesis. Activation of colanic acid synthesis was not dependent on growth at low temperatures (less than or equal to 30 degrees C). The K. pneumoniae rcsA gene complemented E. coli K-12 rcsA mutations but could not complement defects in rcsB, suggesting that RcsA may be functionally homologous in these bacteria. The cloned rcsA gene also complemented a defect in nonmucoid strain K20 derivatives that normally produced only trace amounts of K20 antigen and were unable to assemble a wild-type capsular structure. Mutants that were K20-deficient were not complemented. The K antigen capsule of K. pneumoniae therefore joins a growing list of polysaccharide-synthetic systems in which "RcsA-like" proteins are involved.

Full text

PDF

Images in this article

496Image
on p.496
496Image
on p.496
498Image
on p.498
499Image
on p.499
500Image
on p.500
501Image
on p.501

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. 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]
  3. 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]
  4. 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]
  5. Cryz S. J., Jr, Fürer F., Germanier R. Experimental Klebsiella pneumoniae burn wound sepsis: role of capsular polysaccharide. Infect Immun. 1984 Jan;43(1):440–441. doi: 10.1128/iai.43.1.440-441.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DUGUID J. P., WILKINSON J. F. The influence of cultural conditions on polysaccharide production by Aerobacter aerogenes. J Gen Microbiol. 1953 Oct;9(2):174–189. doi: 10.1099/00221287-9-2-174. [DOI] [PubMed] [Google Scholar]
  7. Ditta G., Schmidhauser T., Yakobson E., Lu P., Liang X. W., Finlay D. R., Guiney D., Helinski D. R. Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid. 1985 Mar;13(2):149–153. doi: 10.1016/0147-619x(85)90068-x. [DOI] [PubMed] [Google Scholar]
  8. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Domenico P., Diedrich D. L., Straus D. C. Extracellular polysaccharide production by Klebsiella pneumoniae and its relationship to virulence. Can J Microbiol. 1985 May;31(5):472–478. doi: 10.1139/m85-088. [DOI] [PubMed] [Google Scholar]
  10. Domenico P., Johanson W. G., Jr, Straus D. C. Lobar pneumonia in rats produced by clinical isolates of Klebsiella pneumoniae. Infect Immun. 1982 Jul;37(1):327–335. doi: 10.1128/iai.37.1.327-335.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garegg P. J., Lindberg B., Onn T., Sutherland I. W. Comparative structural studies on the M-antigen from Salmonella typhimurium. Escherichia coli and Aerobacter cloacae. Acta Chem Scand. 1971;25(6):2103–2108. doi: 10.3891/acta.chem.scand.25-2103. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Grant W. D., Sutherland I. W., Wilkinson J. F. Exopolysaccharide colanic acid and its occurrence in the Enterobacteriaceae. J Bacteriol. 1969 Dec;100(3):1187–1193. doi: 10.1128/jb.100.3.1187-1193.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Homonylo M. K., Wilmot S. J., Lam J. S., MacDonald L. A., Whitfield C. Monoclonal antibodies against the capsular K antigen of Escherichia coli (O9:K30(A):H12): characterisation and use in analysis of K antigen organisation on the cell surface. Can J Microbiol. 1988 Oct;34(10):1159–1165. doi: 10.1139/m88-204. [DOI] [PubMed] [Google Scholar]
  16. Hull R. A., Gill R. E., Hsu P., Minshew B. H., Falkow S. Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate. Infect Immun. 1981 Sep;33(3):933–938. doi: 10.1128/iai.33.3.933-938.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Knauf V. C., Nester E. W. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid. 1982 Jul;8(1):45–54. doi: 10.1016/0147-619x(82)90040-3. [DOI] [PubMed] [Google Scholar]
  18. Laakso D. H., Homonylo M. K., Wilmot S. J., Whitfield C. Transfer and expression of the genetic determinants for O and K antigen synthesis in Escherichia coli O9:K(A)30 and Klebsiella sp. O1:K20, in Escherichia coli K12. Can J Microbiol. 1988 Aug;34(8):987–992. doi: 10.1139/m88-173. [DOI] [PubMed] [Google Scholar]
  19. McCallum K. L., Schoenhals G., Laakso D., Clarke B., Whitfield C. A high-molecular-weight fraction of smooth lipopolysaccharide in Klebsiella serotype O1:K20 contains a unique O-antigen epitope and determines resistance to nonspecific serum killing. Infect Immun. 1989 Dec;57(12):3816–3822. doi: 10.1128/iai.57.12.3816-3822.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Montgomerie J. Z. Epidemiology of Klebsiella and hospital-associated infections. Rev Infect Dis. 1979 Sep-Oct;1(5):736–753. doi: 10.1093/clinids/1.5.736. [DOI] [PubMed] [Google Scholar]
  22. Nakashima I., Kobayashi T., Kato N. Alterations in the antibody response to bovine serum albumin by capsular polysaccharide of Klebsiella pneumoniae. J Immunol. 1971 Oct;107(4):1112–1121. [PubMed] [Google Scholar]
  23. Nassif X., Fournier J. M., Arondel J., Sansonetti P. J. Mucoid phenotype of Klebsiella pneumoniae is a plasmid-encoded virulence factor. Infect Immun. 1989 Feb;57(2):546–552. doi: 10.1128/iai.57.2.546-552.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. ORSKOV I. Immunological paralysis induced in rabbits by a heavily capsulated Klebsiella strain. Acta Pathol Microbiol Scand. 1956;38(5):375–384. [PubMed] [Google Scholar]
  26. Pollack M. Significance of circulating capsular antigen in Klebsiella infections. Infect Immun. 1976 Jun;13(6):1543–1548. doi: 10.1128/iai.13.6.1543-1548.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. 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]
  29. 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]
  30. Takahashi M., Yoshida K., San Clemente C. L. Relation of colonial morphologies in soft agar to morphological and biological properties of the K-9 strain of Klebsiella pneumoniae and its variants. Can J Microbiol. 1977 Apr;23(4):448–451. doi: 10.1139/m77-066. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Torres-Cabassa A., Gottesman S., Frederick R. D., Dolph P. J., Coplin D. L. Control of extracellular polysaccharide synthesis in Erwinia stewartii and Escherichia coli K-12: a common regulatory function. J Bacteriol. 1987 Oct;169(10):4525–4531. doi: 10.1128/jb.169.10.4525-4531.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Troy F. A., Frerman F. E., Heath E. C. The biosynthesis of capsular polysaccharide in Aerobacter aerogenes. J Biol Chem. 1971 Jan 10;246(1):118–133. [PubMed] [Google Scholar]
  35. Whitfield C., Vimr E. R., Costerton J. W., Troy F. A. Protein synthesis is required for in vivo activation of polysialic acid capsule synthesis in Escherichia coli K1. J Bacteriol. 1984 Jul;159(1):321–328. doi: 10.1128/jb.159.1.321-328.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Williams P., Lambert P. A., Brown M. R., Jones R. J. The role of the O and K antigens in determining the resistance of Klebsiella aerogenes to serum killing and phagocytosis. J Gen Microbiol. 1983 Jul;129(7):2181–2191. doi: 10.1099/00221287-129-7-2181. [DOI] [PubMed] [Google Scholar]
  37. Williams P., Lambert P. A., Haigh C. G., Brown M. R. The influence of the O and K antigens of Klebsiella aerogenes on surface hydrophobicity and susceptibility to phagocytosis and antimicrobial agents. J Med Microbiol. 1986 Mar;21(2):125–132. doi: 10.1099/00222615-21-2-125. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Yokochi T., Nakashima I., Kato N. Effect of capsular polysaccharide of Klebsiella pneumoniae on the differentiation and functional capacity of macrophages cultured in vitro. Microbiol Immunol. 1977 Oct 20;21(10):601–610. doi: 10.1111/j.1348-0421.1977.tb00328.x. [DOI] [PubMed] [Google Scholar]
  40. Yokochi T., Nakashima I., Kato N. Further studies on generation of macrophages in in vitro cultures of mouse spleen cells and its inhibition by the capsular polysaccharide of Klebsiella pneumoniae. Microbiol Immunol. 1979;23(6):487–499. doi: 10.1111/j.1348-0421.1979.tb00488.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES