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. 1993 Dec;175(23):7515–7522. doi: 10.1128/jb.175.23.7515-7522.1993

Formation of the K30 (group I) capsule in Escherichia coli O9:K30 does not require attachment to lipopolysaccharide lipid A-core.

P R MacLachlan 1, W J Keenleyside 1, C Dodgson 1, C Whitfield 1
PMCID: PMC206907  PMID: 8244920

Abstract

Escherichia coli K antigens (capsular polysaccharides) are divided into two broad classes, designated groups I and II, on the basis of a number of chemical, physical, and genetic criteria. Group I K antigens can be further subdivided on the basis of the absence (group IA) or presence (group IB) of amino sugars in the repeating unit of the K antigen. One criterion proposed for inclusion in group I is covalent linkage of the capsular polysaccharide to the lipid A-core of lipopolysaccharide (LPS). E. coli O9:K30 is a strain with a representative group IA K antigen. This organism synthesizes an LPS-associated low-molecular-weight form of K30 antigen which is called K(LPS). To determine the involvement of LPS lipid A-core in expression of the K30 capsular polysaccharide, E. coli K30/K-12 hybrid strains were constructed with mutations in the E. coli K-12 rfa locus, responsible for the biosynthesis of the LPS core oligosaccharide. These strains lack K(LPS), indicating that a full-length core is required for K(LPS) expression. However, formation of a K30 capsule was unaffected by rfa defects, indicating that attachment to lipid A-core is not an obligatory step for either export of high-molecular-weight capsular polysaccharide or maintenance of the capsular structure on the cell surface. Silver-stained tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles of lipopolysaccharides from other E. coli K serotypes showed that all strains with group IB K antigens expressed some K(LPS). In contrast, some strains with group IA K antigens appear to lack K(LPS). Consequently, although association of group 1 K antigens with lipid A-core is common, it is not a universal marker for inclusion in group I.

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Selected References

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  1. Beynon L. M., Dumanski A. J., McLean R. J., MacLean L. L., Richards J. C., Perry M. B. Capsule structure of Proteus mirabilis (ATCC 49565). J Bacteriol. 1992 Apr;174(7):2172–2177. doi: 10.1128/jb.174.7.2172-2177.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chakraborty A. K., Friebolin H., Stirm S. Primary structure of the Escherichia coli serotype K30 capsular polysaccharide. J Bacteriol. 1980 Feb;141(2):971–972. doi: 10.1128/jb.141.2.971-972.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clarke A. J., Sarabia V., Keenleyside W., MacLachlan P. R., Whitfield C. The compositional analysis of bacterial extracellular polysaccharides by high-performance anion-exchange chromatography. Anal Biochem. 1991 Nov 15;199(1):68–74. doi: 10.1016/0003-2697(91)90270-4. [DOI] [PubMed] [Google Scholar]
  4. Frosch M., Müller A. Phospholipid substitution of capsular polysaccharides and mechanisms of capsule formation in Neisseria meningitidis. Mol Microbiol. 1993 May;8(3):483–493. doi: 10.1111/j.1365-2958.1993.tb01592.x. [DOI] [PubMed] [Google Scholar]
  5. Gibb A. P., Barclay G. R., Poxton I. R., di Padova F. Frequencies of lipopolysaccharide core types among clinical isolates of Escherichia coli defined with monoclonal antibodies. J Infect Dis. 1992 Nov;166(5):1051–1057. doi: 10.1093/infdis/166.5.1051. [DOI] [PubMed] [Google Scholar]
  6. Goldman R. C., Joiner K., Leive L. Serum-resistant mutants of Escherichia coli O111 contain increased lipopolysaccharide, lack an O antigen-containing capsule, and cover more of their lipid A core with O antigen. J Bacteriol. 1984 Sep;159(3):877–882. doi: 10.1128/jb.159.3.877-882.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goldman R. C., Leive L. Heterogeneity of antigenic-side-chain length in lipopolysaccharide from Escherichia coli 0111 and Salmonella typhimurium LT2. Eur J Biochem. 1980;107(1):145–153. doi: 10.1111/j.1432-1033.1980.tb04635.x. [DOI] [PubMed] [Google Scholar]
  8. Goldman R. C., White D., Orskov F., Orskov I., Rick P. D., Lewis M. S., Bhattacharjee A. K., Leive L. A surface polysaccharide of Escherichia coli O111 contains O-antigen and inhibits agglutination of cells by O-antiserum. J Bacteriol. 1982 Sep;151(3):1210–1221. doi: 10.1128/jb.151.3.1210-1221.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gotschlich E. C., Fraser B. A., Nishimura O., Robbins J. B., Liu T. Y. Lipid on capsular polysaccharides of gram-negative bacteria. J Biol Chem. 1981 Sep 10;256(17):8915–8921. [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. Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Holst O., Zähringer U., Brade H., Zamojski A. Structural analysis of the heptose/hexose region of the lipopolysaccharide from Escherichia coli K-12 strain W3100. Carbohydr Res. 1991 Aug 20;215(2):323–335. doi: 10.1016/0008-6215(91)84031-9. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hungerer D., Jann K., Jann B., Orskov F., Orskov I. Immunochemistry of K antigens of Escherichia coli. 4. The K antigen of E. coli O 9:K30:H12. Eur J Biochem. 1967 Jul;2(1):115–126. doi: 10.1111/j.1432-1033.1967.tb00115.x. [DOI] [PubMed] [Google Scholar]
  15. Jann B., Jann K. Structure and biosynthesis of the capsular antigens of Escherichia coli. Curr Top Microbiol Immunol. 1990;150:19–42. doi: 10.1007/978-3-642-74694-9_2. [DOI] [PubMed] [Google Scholar]
  16. Jann K., Dengler T., Jann B. Core-lipid A on the K40 polysaccharide of Escherichia coli O8:K40:H9, a representative of group I capsular polysaccharides. Zentralbl Bakteriol. 1992 Jan;276(2):196–204. doi: 10.1016/s0934-8840(11)80006-x. [DOI] [PubMed] [Google Scholar]
  17. Jann K., Jann B. Capsules of Escherichia coli, expression and biological significance. Can J Microbiol. 1992 Jul;38(7):705–710. doi: 10.1139/m92-116. [DOI] [PubMed] [Google Scholar]
  18. Jann K., Jann B. The K antigens of Escherichia coli. Prog Allergy. 1983;33:53–79. doi: 10.1159/000407421. [DOI] [PubMed] [Google Scholar]
  19. Jansson P. E., Lindberg A. A., Lindberg B., Wollin R. Structural studies on the hexose region of the core in lipopolysaccharides from Enterobacteriaceae. Eur J Biochem. 1981 Apr;115(3):571–577. doi: 10.1111/j.1432-1033.1981.tb06241.x. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Johnson K. G., Perry M. B. Improved techniques for the preparation of bacterial lipopolysaccharides. Can J Microbiol. 1976 Jan;22(1):29–34. doi: 10.1139/m76-004. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Kiss P., Rinno J., Schmidt G., Mayer H. Structural studies on the immunogenic form of the enterobacterial common antigen. Eur J Biochem. 1978 Jul 17;88(1):211–218. doi: 10.1111/j.1432-1033.1978.tb12440.x. [DOI] [PubMed] [Google Scholar]
  24. Klena J. D., Ashford R. S., 2nd, Schnaitman C. A. Role of Escherichia coli K-12 rfa genes and the rfp gene of Shigella dysenteriae 1 in generation of lipopolysaccharide core heterogeneity and attachment of O antigen. J Bacteriol. 1992 Nov;174(22):7297–7307. doi: 10.1128/jb.174.22.7297-7307.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Kuhn H. M., Meier-Dieter U., Mayer H. ECA, the enterobacterial common antigen. FEMS Microbiol Rev. 1988 Sep;4(3):195–222. doi: 10.1111/j.1574-6968.1988.tb02743.x. [DOI] [PubMed] [Google Scholar]
  27. Kuhn H. M., Neter E., Mayer H. Modification of the lipid moiety of the enterobacterial common antigen by the "Pseudomonas factor". Infect Immun. 1983 May;40(2):696–700. doi: 10.1128/iai.40.2.696-700.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kuo J. S., Doelling V. W., Graveline J. F., McCoy D. W. Evidence for covalent attachment of phospholipid to the capsular polysaccharide of Haemophilus influenzae type b. J Bacteriol. 1985 Aug;163(2):769–773. doi: 10.1128/jb.163.2.769-773.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Lesse A. J., Campagnari A. A., Bittner W. E., Apicella M. A. Increased resolution of lipopolysaccharides and lipooligosaccharides utilizing tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Immunol Methods. 1990 Jan 24;126(1):109–117. doi: 10.1016/0022-1759(90)90018-q. [DOI] [PubMed] [Google Scholar]
  31. Low K. B. Conjugational methods for mapping with Hfr and F-prime strains. Methods Enzymol. 1991;204:43–62. doi: 10.1016/0076-6879(91)04005-9. [DOI] [PubMed] [Google Scholar]
  32. MacIntyre S., Lucken R., Owen P. Smooth lipopolysaccharide is the major protective antigen for mice in the surface extract from IATS serotype 6 contributing to the polyvalent Pseudomonas aeruginosa vaccine PEV. Infect Immun. 1986 Apr;52(1):76–84. doi: 10.1128/iai.52.1.76-84.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. McCallum K. L., Laakso D. H., Whitfield C. Use of a bacteriophage-encoded glycanase enzyme in the generation of lipopolysaccharide O side chain deficient mutants of Escherichia coli O9:K30 and Klebsiella O1:K20: role of O and K antigens in resistance to complement-mediated serum killing. Can J Microbiol. 1989 Nov;35(11):994–999. doi: 10.1139/m89-166. [DOI] [PubMed] [Google Scholar]
  34. McGrath B. C., Osborn M. J. Localization of the terminal steps of O-antigen synthesis in Salmonella typhimurium. J Bacteriol. 1991 Jan;173(2):649–654. doi: 10.1128/jb.173.2.649-654.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Munford R. S., Hall C. L., Rick P. D. Size heterogeneity of Salmonella typhimurium lipopolysaccharides in outer membranes and culture supernatant membrane fragments. J Bacteriol. 1980 Nov;144(2):630–640. doi: 10.1128/jb.144.2.630-640.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nikaido H., Vaara M. Molecular basis of bacterial outer membrane permeability. Microbiol Rev. 1985 Mar;49(1):1–32. doi: 10.1128/mr.49.1.1-32.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Orskov I., Orskov F., Jann B., Jann K. Serology, chemistry, and genetics of O and K antigens of Escherichia coli. Bacteriol Rev. 1977 Sep;41(3):667–710. doi: 10.1128/br.41.3.667-710.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Palva E. T., Mäkelä P. H. Lipopolysaccharide heterogeneity in Salmonella typhimurium analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Eur J Biochem. 1980;107(1):137–143. doi: 10.1111/j.1432-1033.1980.tb04634.x. [DOI] [PubMed] [Google Scholar]
  39. Parker C. T., Kloser A. W., Schnaitman C. A., Stein M. A., Gottesman S., Gibson B. W. Role of the rfaG and rfaP genes in determining the lipopolysaccharide core structure and cell surface properties of Escherichia coli K-12. J Bacteriol. 1992 Apr;174(8):2525–2538. doi: 10.1128/jb.174.8.2525-2538.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Pelkonen S., Häyrinen J., Finne J. Polyacrylamide gel electrophoresis of the capsular polysaccharides of Escherichia coli K1 and other bacteria. J Bacteriol. 1988 Jun;170(6):2646–2653. doi: 10.1128/jb.170.6.2646-2653.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Peterson A. A., McGroarty E. J. High-molecular-weight components in lipopolysaccharides of Salmonella typhimurium, Salmonella minnesota, and Escherichia coli. J Bacteriol. 1985 May;162(2):738–745. doi: 10.1128/jb.162.2.738-745.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schmidt G., Jann B., Jann K. Immunochemistry of R lipopolysaccharides of Escherichia coli. Different core regions in the lipopolysaccharides of O group 8. Eur J Biochem. 1969 Oct;10(3):501–510. doi: 10.1111/j.1432-1033.1969.tb00717.x. [DOI] [PubMed] [Google Scholar]
  43. Schmidt G., Jann B., Jann K., Orskov I., Orskov F. Genetic determinants of the synthesis of the polysaccharide capsular antigen K27(A) of Escherichia coli. J Gen Microbiol. 1977 Jun;100(2):355–361. doi: 10.1099/00221287-100-2-355. [DOI] [PubMed] [Google Scholar]
  44. Sternberg N. L., Maurer R. Bacteriophage-mediated generalized transduction in Escherichia coli and Salmonella typhimurium. Methods Enzymol. 1991;204:18–43. doi: 10.1016/0076-6879(91)04004-8. [DOI] [PubMed] [Google Scholar]
  45. 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]
  46. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  47. Whitfield C., Schoenhals G., Graham L. Mutants of Escherichia coli O9:K30 with altered synthesis and expression of the capsular K30 antigen. J Gen Microbiol. 1989 Oct;135(10):2589–2599. doi: 10.1099/00221287-135-10-2589. [DOI] [PubMed] [Google Scholar]
  48. 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]
  49. Wilkinson R. G., Gemski P., Jr, Stocker B. A. Non-smooth mutants of Salmonella typhimurium: differentiation by phage sensitivity and genetic mapping. J Gen Microbiol. 1972 May;70(3):527–554. doi: 10.1099/00221287-70-3-527. [DOI] [PubMed] [Google Scholar]

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