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. 1989 Jun;171(6):3074–3079. doi: 10.1128/jb.171.6.3074-3079.1989

Activity of CMP-2-keto-3-deoxyoctulosonic acid synthetase in Escherichia coli strains expressing the capsular K5 polysaccharide implication for K5 polysaccharide biosynthesis.

A Finke 1, I Roberts 1, G Boulnois 1, C Pzzani 1, K Jann 1
PMCID: PMC210017  PMID: 2542215

Abstract

The activity of the cytoplasmic CMP-2-keto-3-deoxyoctulosonic acid synthetase (CMP-KDO synthetase), which is low in Escherichia coli rough strains such as E. coli K-12 and in uncapsulated strains such as E. coli O111, was significantly elevated in encapsulated E. coli O10:K5 and O18:K5. This enzyme activity was even higher in an E. coli clone expressing the K5 capsule. This and the following findings suggest a correlation between elevated CMP-KDO synthetase activity and the biosynthesis of the capsular K5 polysaccharide. (i) Expression of the K5 polysaccharide and elevated CMP-KDO synthetase activity were observed with bacteria grown at 37 degrees C but not with cells grown at 20 degrees C or below. (ii) The recovery kinetics of capsule expression of intact bacteria, in vitro K5 polysaccharide-synthesizing activity of bacteria, and CMP-KDO synthetase activity of bacteria after temperature upshift from 18 to 37 degrees C were the same. (iii) Chemicals which inhibit capsule (polysaccharide) expression also inhibited the elevation of CMP-KDO synthetase activity. The chromosomal location of the gene responsible for the elevation of this enzyme activity was narrowed down to the distal segment of the transport region of the K5 expression genes.

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Selected References

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  1. Boulnois G. J., Roberts I. S., Hodge R., Hardy K. R., Jann K. B., Timmis K. N. Analysis of the K1 capsule biosynthesis genes of Escherichia coli: definition of three functional regions for capsule production. Mol Gen Genet. 1987 Jun;208(1-2):242–246. doi: 10.1007/BF00330449. [DOI] [PubMed] [Google Scholar]
  2. Ghalambor M. A., Heath E. C. The biosynthesis of cell wall lipopolysaccharide in Escherichia coli. IV. Purification and properties of cytidine monophosphate 3-deoxy-d-manno-octulosonate synthetase. J Biol Chem. 1966 Jul 10;241(13):3216–3221. [PubMed] [Google Scholar]
  3. 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]
  4. Jann K., Jann B. The K antigens of Escherichia coli. Prog Allergy. 1983;33:53–79. doi: 10.1159/000407421. [DOI] [PubMed] [Google Scholar]
  5. Orskov F., Sharma V., Orskov I. Influence of growth temperature on the development of Escherichia coli polysaccharide K antigens. J Gen Microbiol. 1984 Oct;130(10):2681–2684. doi: 10.1099/00221287-130-10-2681. [DOI] [PubMed] [Google Scholar]
  6. Orskov I., Nyman K. Genetic mapping of the antigenic determinants of two polysaccharide K antigens, K10 and K54, in Escherichia coli. J Bacteriol. 1974 Oct;120(1):43–51. doi: 10.1128/jb.120.1.43-51.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Orskov I., Sharma V., Orskov F. Genetic mapping of the K1 and K4 antigens (L) of Escherichia coli. Non-allelism of K(L) antigens with K antigens of O8:K27(A), O8:K8(L) and O9:K57(B). Acta Pathol Microbiol Scand B. 1976 Jun;84(3):125–131. [PubMed] [Google Scholar]
  9. Osborn M. J., Rick P. D., Rasmussen N. S. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Translocation and integration of an incomplete mutant lipid A into the outer membrane. J Biol Chem. 1980 May 10;255(9):4246–4251. [PubMed] [Google Scholar]
  10. Prihar H. S., Campbell P., Feingold D. S., Jacobsson I., Jensen J. W., Lindahl U., Rodén L. Biosynthesis of heparin. Hydrogen exchange at carbon 5 of the glucuronosyl residues. Biochemistry. 1980 Feb 5;19(3):495–500. doi: 10.1021/bi00544a016. [DOI] [PubMed] [Google Scholar]
  11. Raetz C. R., Purcell S., Meyer M. V., Qureshi N., Takayama K. Isolation and characterization of eight lipid A precursors from a 3-deoxy-D-manno-octylosonic acid-deficient mutant of Salmonella typhimurium. J Biol Chem. 1985 Dec 25;260(30):16080–16088. [PubMed] [Google Scholar]
  12. Ray P. H., Benedict C. D. CTP:CMP-3-deoxy-D-manno-octulosonate cytidylyltransferase (CMP-KDO synthetase). Methods Enzymol. 1982;83:535–540. doi: 10.1016/0076-6879(82)83049-8. [DOI] [PubMed] [Google Scholar]
  13. Rick P. D., Fung L. W., Ho C., Osborn M. J. Lipid A mutants of Salmonella typhimurium. Purification and characterization of a lipid A precursor produced by a mutant in 3-deoxy-D-mannooctulosonate-8-phosphate synthetase. J Biol Chem. 1977 Jul 25;252(14):4904–4912. [PubMed] [Google Scholar]
  14. Roberts I. S., Mountford R., Hodge R., Jann K. B., Boulnois G. J. Common organization of gene clusters for production of different capsular polysaccharides (K antigens) in Escherichia coli. J Bacteriol. 1988 Mar;170(3):1305–1310. doi: 10.1128/jb.170.3.1305-1310.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Roberts I., Mountford R., High N., Bitter-Suermann D., Jann K., Timmis K., Boulnois G. Molecular cloning and analysis of genes for production of K5, K7, K12, and K92 capsular polysaccharides in Escherichia coli. J Bacteriol. 1986 Dec;168(3):1228–1233. doi: 10.1128/jb.168.3.1228-1233.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Vann W. F., Schmidt M. A., Jann B., Jann K. The structure of the capsular polysaccharide (K5 antigen) of urinary-tract-infective Escherichia coli 010:K5:H4. A polymer similar to desulfo-heparin. Eur J Biochem. 1981 May 15;116(2):359–364. doi: 10.1111/j.1432-1033.1981.tb05343.x. [DOI] [PubMed] [Google Scholar]
  17. Westphal O., Jann K., Himmelspach K. Chemistry and immunochemistry of bacterial lipopolysaccharides as cell wall antigens and endotoxins. Prog Allergy. 1983;33:9–39. [PubMed] [Google Scholar]
  18. 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]

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