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. 1988 Jun;170(6):2599–2611. doi: 10.1128/jb.170.6.2599-2611.1988

Fine-structure mapping and identification of two regulators of capsule synthesis in Escherichia coli K-12.

J A Brill 1, C Quinlan-Walshe 1, S Gottesman 1
PMCID: PMC211177  PMID: 2836365

Abstract

Positive and negative regulatory elements involved in the synthesis of colanic acid, the capsular polysaccharide of Escherichia coli K-12, have been identified previously. RcsB, a positive regulator for transcription of the structural genes of colanic acid synthesis (cps), is a protein of about 26 kilodaltons which probably acts as a multimer, rcsC, which maps close to rcsB at 48 min on the E. coli chromosome, exerts a negative effect on expression of the structural genes and codes for a protein of about 100 kilodaltons. The two genes appear to be transcribed in opposite directions, with the C-terminal ends of the genes being less than 0.3 kilobases apart. Multicopy expression of rcsB is lethal in rcsC mutants which carry cps-lac fusions, probably owing to accumulation of intermediates in the capsule synthesis pathway in these cells. Examination of double mutants and cells carrying multicopy rcsB+ plasmids reveal an rcsA-independent pathway for capsule synthesis. We hypothesize that RcsC may act as an environmental sensor, transmitting information to the RcsB positive regulator.

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Selected References

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  1. Allen P. M., Fisher D., Saunders J. R., Hart C. A. The role of capsular polysaccharide K21b of Klebsiella and of the structurally related colanic-acid polysaccharide of Escherichia coli in resistance to phagocytosis and serum killing. J Med Microbiol. 1987 Dec;24(4):363–370. doi: 10.1099/00222615-24-4-363. [DOI] [PubMed] [Google Scholar]
  2. Backman K. C., Chen Y. M., Ueno-Nishio S., Magasanik B. The product of glnL is not essential for regulation of bacterial nitrogen assimilation. J Bacteriol. 1983 Apr;154(1):516–519. doi: 10.1128/jb.154.1.516-519.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradshaw-Rouse J. J., Whatley M. H., Coplin D. L., Woods A., Sequeira L., Kelman A. Agglutination of Erwinia stewartii Strains with a Corn Agglutinin: Correlation with Extracellular Polysaccharide Production and Pathogenicity. Appl Environ Microbiol. 1981 Aug;42(2):344–350. doi: 10.1128/aem.42.2.344-350.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bueno R., Pahel G., Magasanik B. Role of glnB and glnD gene products in regulation of the glnALG operon of Escherichia coli. J Bacteriol. 1985 Nov;164(2):816–822. doi: 10.1128/jb.164.2.816-822.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Charette M. F., Henderson G. W., Markovitz A. ATP hydrolysis-dependent protease activity of the lon (capR) protein of Escherichia coli K-12. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4728–4732. doi: 10.1073/pnas.78.8.4728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chattoraj D. K., Cordes K., Berman M. L., Das A. Mutagenesis and mutation transfer induced by ultraviolet light in plasmid-cloned DNA. Gene. 1984 Feb;27(2):213–222. doi: 10.1016/0378-1119(84)90142-2. [DOI] [PubMed] [Google Scholar]
  7. Chung C. H., Goldberg A. L. The product of the lon (capR) gene in Escherichia coli is the ATP-dependent protease, protease La. Proc Natl Acad Sci U S A. 1981 Aug;78(8):4931–4935. doi: 10.1073/pnas.78.8.4931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coplin D. L., Frederick R. D., Majerczak D. R., Haas E. S. Molecular cloning of virulence genes from Erwinia stewartii. J Bacteriol. 1986 Nov;168(2):619–623. doi: 10.1128/jb.168.2.619-623.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Costerton J. W., Irvin R. T., Cheng K. J. The role of bacterial surface structures in pathogenesis. Crit Rev Microbiol. 1981;8(4):303–338. doi: 10.3109/10408418109085082. [DOI] [PubMed] [Google Scholar]
  10. Goldberg A. L., Swamy K. H., Chung C. H., Larimore F. S. Proteases in Escherichia coli. Methods Enzymol. 1981;80(Pt 100):680–702. doi: 10.1016/s0076-6879(81)80052-3. [DOI] [PubMed] [Google Scholar]
  11. Gottesman S., Gottesman M., Shaw J. E., Pearson M. L. Protein degradation in E. coli: the lon mutation and bacteriophage lambda N and cII protein stability. Cell. 1981 Apr;24(1):225–233. doi: 10.1016/0092-8674(81)90518-3. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Gottesman S., Zipser D. Deg phenotype of Escherichia coli lon mutants. J Bacteriol. 1978 Feb;133(2):844–851. doi: 10.1128/jb.133.2.844-851.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. HOWARD-FLANDERS P., SIMSON E., THERIOT L. A LOCUS THAT CONTROLS FILAMENT FORMATION AND SENSITIVITY TO RADIATION IN ESCHERICHIA COLI K-12. Genetics. 1964 Feb;49:237–246. doi: 10.1093/genetics/49.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kanaya S., Crouch R. J. The rnh gene is essential for growth of Escherichia coli. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3447–3451. doi: 10.1073/pnas.81.11.3447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Leigh J. A., Signer E. R., Walker G. C. Exopolysaccharide-deficient mutants of Rhizobium meliloti that form ineffective nodules. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6231–6235. doi: 10.1073/pnas.82.18.6231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Magasanik B. Genetic control of nitrogen assimilation in bacteria. Annu Rev Genet. 1982;16:135–168. doi: 10.1146/annurev.ge.16.120182.001031. [DOI] [PubMed] [Google Scholar]
  20. Maloy S. R., Nunn W. D. Selection for loss of tetracycline resistance by Escherichia coli. J Bacteriol. 1981 Feb;145(2):1110–1111. doi: 10.1128/jb.145.2.1110-1111.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maurer R., Osmond B. C., Shekhtman E., Wong A., Botstein D. Functional interchangeability of DNA replication genes in Salmonella typhimurium and Escherichia coli demonstrated by a general complementation procedure. Genetics. 1984 Sep;108(1):1–23. doi: 10.1093/genetics/108.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Mizuno T., Chou M. Y., Inouye M. A comparative study on the genes for three porins of the Escherichia coli outer membrane. DNA sequence of the osmoregulated ompC gene. J Biol Chem. 1983 Jun 10;258(11):6932–6940. [PubMed] [Google Scholar]
  24. Mizuno T., Chou M. Y., Inouye M. DNA sequence of the promoter region of the ompC gene and the amino acid sequence of the signal peptide of pro-OmpC protein of Escherichia coli. FEBS Lett. 1983 Jan 10;151(1):159–164. doi: 10.1016/0014-5793(83)80364-0. [DOI] [PubMed] [Google Scholar]
  25. Mizusawa S., Gottesman S. Protein degradation in Escherichia coli: the lon gene controls the stability of sulA protein. Proc Natl Acad Sci U S A. 1983 Jan;80(2):358–362. doi: 10.1073/pnas.80.2.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mizusawa S., Ward D. F. A bacteriophage lambda vector for cloning with BamHI and Sau3A. Gene. 1982 Dec;20(3):317–322. doi: 10.1016/0378-1119(82)90200-1. [DOI] [PubMed] [Google Scholar]
  27. Nara F., Mizuno T., Mizushima S. Complementation analysis of the wild-type and mutant ompR genes exhibiting different phenotypes of osmoregulation of the ompF and ompC genes of Escherichia coli. Mol Gen Genet. 1986 Oct;205(1):51–55. [PubMed] [Google Scholar]
  28. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ninfa A. J., Magasanik B. Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5909–5913. doi: 10.1073/pnas.83.16.5909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nixon B. T., Ronson C. W., Ausubel F. M. Two-component regulatory systems responsive to environmental stimuli share strongly conserved domains with the nitrogen assimilation regulatory genes ntrB and ntrC. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7850–7854. doi: 10.1073/pnas.83.20.7850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Slauch J. M., Garrett S., Jackson D. E., Silhavy T. J. EnvZ functions through OmpR to control porin gene expression in Escherichia coli K-12. J Bacteriol. 1988 Jan;170(1):439–441. doi: 10.1128/jb.170.1.439-441.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. 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]
  35. Way J. C., Davis M. A., Morisato D., Roberts D. E., Kleckner N. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene. 1984 Dec;32(3):369–379. doi: 10.1016/0378-1119(84)90012-x. [DOI] [PubMed] [Google Scholar]
  36. Wyman A. R., Wolfe L. B., Botstein D. Propagation of some human DNA sequences in bacteriophage lambda vectors requires mutant Escherichia coli hosts. Proc Natl Acad Sci U S A. 1985 May;82(9):2880–2884. doi: 10.1073/pnas.82.9.2880. [DOI] [PMC free article] [PubMed] [Google Scholar]

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