Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1973 Sep;115(3):1011–1020. doi: 10.1128/jb.115.3.1011-1020.1973

Derepression of Uridine Diphosphate-Glucose Pyrophosphorylase (galU) in capR(lon), capS, and capT Mutants and Studies on the galU Repressor

Christine E Buchanan a,1, Alvin Markovitz a
PMCID: PMC246349  PMID: 4580555

Abstract

Mutation of the capR(lon), capS, or capT genes in Escherichia coli K-12 causes overproduction of capsular polysaccharide leading to a mucoid phenotype. Several of the enzymes involved in capsular polysaccharide synthesis are derepressed in cap mutants. Previously it was shown that uridine diphosphate-glucose (UDPG) pyrophosphorylase, an enzyme involved in the synthesis of three of the nucleotide sugar precursors of the capsule, is derepressed in capR mutants. The control of galU, the gene which codes for UDPG pyrophosphorylase, is described in this study. In addition, it has been found that the enzyme is also derepressed in capS and capT mutants. The effect of galU gene dosage in cap mutants and the wild-type strain (all lysogenic for φ80) was studied by infecting them with the purified transducing phage φ80dgalU. The level of UDPG pyrophosphorylase increased in proportion to the number of galU copies added. The rate of enzyme synthesis in the mutants was about sixfold higher than in the wild type per galU gene added for multiplicities of infection from one to twenty. Thus, all the galU copies added to the wild-type lysogen were repressed. We obtain greater than 20 galU copies per cell by infecting the nonlysogenic strain which allows multiplication of φ80dgalU. With some number of galU copies greater than 20, the rate of UDPG pyrophosphorylase synthesis in the wild type approaches the mutant rate of synthesis. The results suggest that there may indeed be a galU repressor pool in the cell which can be completely titrated. This pool must be composed of more than 20 galU repressor molecules. Since the capR, capS, and capT gene products or combinations thereof are known to control other widely separated operons of the cell besides the galU gene, it is postulated that the galU repressor may be capable of binding other operators. This would account for the relatively large pool of galU repressors per cell.

Full text

PDF
1011

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ADLER H. I., HARDIGREE A. A. ANALYSIS OF A GENE CONTROLLING CELL DIVISION AND SENSITIVITY TO RADIATION IN ESCHERICHIA COLI. J Bacteriol. 1964 Mar;87:720–726. doi: 10.1128/jb.87.3.720-726.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BREITMAN T. R. The feedback inhibition of thymidine kinase. Biochim Biophys Acta. 1963 Jan 8;67:153–155. doi: 10.1016/0006-3002(63)91807-9. [DOI] [PubMed] [Google Scholar]
  3. BUTTIN G., JACOB F., MONOD J. [Constituent synthesis of galactokinase following the development of lambda bacteriophages in Escherichia coli K 12]. C R Hebd Seances Acad Sci. 1960 Mar 28;250:2471–2473. [PubMed] [Google Scholar]
  4. BUTTIN G. M'ECANISMES R'EGULATEURS DANS LA BIOSYNTH'ESE DES ENZYMES DU M'ETABOLISME DU GALACTOSE CHEZ ESCHERICHIA COLI K12. III. L'"EFFET DE D'ER'EPRESSION" PROVOQU'E PAR LE D'EVELOPPEMENT DU PHAGE LAMBDA. J Mol Biol. 1963 Dec;7:610–631. doi: 10.1016/s0022-2836(63)80108-4. [DOI] [PubMed] [Google Scholar]
  5. BUTTIN G. Some aspects of regulation in the synthesis of the enzymes governing galactose metabolism. Cold Spring Harb Symp Quant Biol. 1961;26:213–216. doi: 10.1101/sqb.1961.026.01.025. [DOI] [PubMed] [Google Scholar]
  6. Beckwith J. R., Signer E. R., Epstein W. Transposition of the Lac region of E. coli. Cold Spring Harb Symp Quant Biol. 1966;31:393–401. doi: 10.1101/sqb.1966.031.01.051. [DOI] [PubMed] [Google Scholar]
  7. Buchanan C. E., Hua S. S., Avni H., Markovitz A. Transcriptional control of the calactose operon by the capR (lon) and capT genes. J Bacteriol. 1973 May;114(2):891–893. doi: 10.1128/jb.114.2.891-893.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bush J. W., Markovitz A. The Genetic Basis for Mucoidy and Radiation Sensitivity in capR (lon) Mutants of E. coli K-12. Genetics. 1973 Jun;74(2):215–225. doi: 10.1093/genetics/74.2.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Calendar R. The regulation of phage development. Annu Rev Microbiol. 1970;24:241–296. doi: 10.1146/annurev.mi.24.100170.001325. [DOI] [PubMed] [Google Scholar]
  10. Deeb S. S. Morphogenesis of bacteriophage phi 80: identification of the cistron 13 product. J Virol. 1972 Jan;9(1):174–181. doi: 10.1128/jvi.9.1.174-181.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Epstein W. Transposition of the lac region of Escherichia coli. IV. Escape from repression in bacteriophage-carried lac genes. J Mol Biol. 1967 Dec 28;30(3):529–543. doi: 10.1016/0022-2836(67)90366-x. [DOI] [PubMed] [Google Scholar]
  12. FUKASAWA T., JOKURA K., KURAHASHI K. A new enzymic defect of galactose metabolism in Escherichia coli K-12 mutants. Biochem Biophys Res Commun. 1962 Apr 3;7:121–125. doi: 10.1016/0006-291x(62)90158-4. [DOI] [PubMed] [Google Scholar]
  13. Germaine G. R., Rogers P. Role of gal repressor depletion in lambda-dg transduction escape synthesis. J Mol Biol. 1970 Jan 28;47(2):121–135. doi: 10.1016/0022-2836(70)90334-7. [DOI] [PubMed] [Google Scholar]
  14. Gilbert W., Müller-Hill B. Isolation of the lac repressor. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1891–1898. doi: 10.1073/pnas.56.6.1891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gilbert W., Müller-Hill B. The lac operator is DNA. Proc Natl Acad Sci U S A. 1967 Dec;58(6):2415–2421. doi: 10.1073/pnas.58.6.2415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. HERSHEY A. D. An upper limit to the protein content of the germinal substance of bacteriophage T2. Virology. 1955 May;1(1):108–127. doi: 10.1016/0042-6822(55)90009-x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Hiraga S., Igarashi K., Yura T. A deoxythymidine kinase-deficient mutant of Escherichia coli. I. Isolation and some properties. Biochim Biophys Acta. 1967 Aug 22;145(1):41–51. doi: 10.1016/0005-2787(67)90652-1. [DOI] [PubMed] [Google Scholar]
  19. Hua S. S., Markovitz A. Multiple regulator gene control of the galactose operon in Escherichia coli K-12. J Bacteriol. 1972 Jun;110(3):1089–1099. doi: 10.1128/jb.110.3.1089-1099.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Igarashi K., Hiraga S., Yura T. A deoxythymidine kinase deficient mutant of Escherichia coli. II. Mapping and transduction studies with phage phi 80. Genetics. 1967 Nov;57(3):643–654. doi: 10.1093/genetics/57.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Inokuchi H., Ozeki H. Phenotypic mixing between bacteriophage phi80 and lambda in vitro and in vivo. Virology. 1970 Aug;41(4):701–710. doi: 10.1016/0042-6822(70)90434-4. [DOI] [PubMed] [Google Scholar]
  22. Koch A. L. The tight binding nature of the repressor-operon interaction. J Theor Biol. 1967 Aug;16(2):166–186. doi: 10.1016/0022-5193(67)90003-3. [DOI] [PubMed] [Google Scholar]
  23. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  24. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  25. Lieberman M. M., Buchanan C. E., Markovitz A. Derepression of GDP-alpha-mannose and UDP-glucose pyrophosphorylases by a regulator gene mutation; episomal dominance in partial diploids. Proc Natl Acad Sci U S A. 1970 Mar;65(3):625–632. doi: 10.1073/pnas.65.3.625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lieberman M. M., Markovitz A. Depression of guanosine diphosphate-mannose pyrophosphorylase by mutations in two different regulator genes involved in capsular polysaccharide synthesis in Escherichia coli K-12. J Bacteriol. 1970 Mar;101(3):965–972. doi: 10.1128/jb.101.3.965-972.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lieberman M. M., Shaparis A., Markovitz A. Control of uridine diphosphate-glucose dehydrogenase synthesis and uridine diphosphate-glucuronic acid accumulation by a regulator gene mutation in Escherichia coli K-12. J Bacteriol. 1970 Mar;101(3):959–964. doi: 10.1128/jb.101.3.959-964.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. MARKOVITZ A. REGULATORY MECHANISMS FOR SYNTHESIS OF CAPSULAR POLYSACCHARIDE IN MUCOID MUTANTS OF ESCHERICHIA COLI K12. Proc Natl Acad Sci U S A. 1964 Feb;51:239–246. doi: 10.1073/pnas.51.2.239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mackie G., Wilson D. B. Regulation of the gal operon of Escherichia coli by the capR gene. J Biol Chem. 1972 May 25;247(10):2973–2978. [PubMed] [Google Scholar]
  30. Markovitz A., Baker B. Suppression of radiation sensitivity and capsular polysaccharide synthesis in Escherichia coli K-12 by ochre suppressors. J Bacteriol. 1967 Aug;94(2):388–395. doi: 10.1128/jb.94.2.388-395.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Markovitz A., Lieberman M. M., Rosenbaum N. Derepression of phosphomannose isomerase by regulator gene mutations involved in capsular polysaccharide synthesis in Escherichia coli K-12. J Bacteriol. 1967 Nov;94(5):1497–1501. doi: 10.1128/jb.94.5.1497-1501.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Markovitz A., Rosenbaum N. A regulator gene that is dominant on an episome and recessive on a chromosome. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1084–1091. doi: 10.1073/pnas.54.4.1084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Markovitz A., Sydiskis R. J., Lieberman M. M. Genetic and biochemical studies on mannose-negative mutants that are deficient in phosphomannose isomerase in Escherichia coli K-12. J Bacteriol. 1967 Nov;94(5):1492–1496. doi: 10.1128/jb.94.5.1492-1496.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Parks J. S., Gottesman M., Shimada K., Weisberg R. A., Perlman R. L., Pastan I. Isolation of the gal repressor. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1891–1895. doi: 10.1073/pnas.68.8.1891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ptashne M., Hopkins N. The operators controlled by the lambda phage repressor. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1282–1287. doi: 10.1073/pnas.60.4.1282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ptashne M. Specific binding of the lambda phage repressor to lambda DNA. Nature. 1967 Apr 15;214(5085):232–234. doi: 10.1038/214232a0. [DOI] [PubMed] [Google Scholar]
  37. REVEL H. R., LURIA S. E. Biosynthesis of beta-D-galactosidase controlled by phage-carried genes. II. The behavior of phage-transduced z-plus genes toward regulatory mechanisms. Proc Natl Acad Sci U S A. 1961 Dec 15;47:1968–1974. doi: 10.1073/pnas.47.12.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. REVEL H. R., LURIA S. E., ROTMAN B. Biosynthesis of B-D-galactosidase controlled by phage-carried genes. I. Induced beta-D-galactosidase biosynthesis after transduction of gene z-plus by phage. Proc Natl Acad Sci U S A. 1961 Dec 15;47:1956–1967. doi: 10.1073/pnas.47.12.1956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. REVEL H. R., LURIA S. E., YOUNG N. L. Biosynthesis of beta-D-galactosidase controlled by phage-carried genes. III. Dereprssion of beta-d-galactosidase synthesis following induction of phage development in lysogenic bacteria. Proc Natl Acad Sci U S A. 1961 Dec 15;47:1974–1980. doi: 10.1073/pnas.47.12.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Reichardt L., Kaiser A. D. Control of lambda repressor synthesis. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2185–2189. doi: 10.1073/pnas.68.9.2185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Riggs A. D., Bourgeois S., Newby R. F., Cohn M. DNA binding of the lac repressor. J Mol Biol. 1968 Jul 14;34(2):365–368. doi: 10.1016/0022-2836(68)90261-1. [DOI] [PubMed] [Google Scholar]
  42. Riggs A. D., Suzuki H., Bourgeois S. Lac repressor-operator interaction. I. Equilibrium studies. J Mol Biol. 1970 Feb 28;48(1):67–83. doi: 10.1016/0022-2836(70)90219-6. [DOI] [PubMed] [Google Scholar]
  43. Sato K., Matsushiro A. The tryptophan operon regulated by phage immunity. J Mol Biol. 1965 Dec;14(2):608–610. doi: 10.1016/s0022-2836(65)80212-1. [DOI] [PubMed] [Google Scholar]
  44. Sato K., Nishimune Y., Sato M., Numich R., Matsushiro A. Suppressor-sensitive mutants of coliphage phi-80. Virology. 1968 Apr;34(4):637–649. [PubMed] [Google Scholar]
  45. Schleif R., Greenblatt J., Davis R. W. Dual control of arabinose genes on transducing phage lambda-dara. J Mol Biol. 1971 Jul 14;59(1):127–150. doi: 10.1016/0022-2836(71)90417-7. [DOI] [PubMed] [Google Scholar]
  46. Shapiro J. A. Chromosomal location of the gene determining uridine diphoglucose formation in Escherichia coli K-12. J Bacteriol. 1966 Aug;92(2):518–520. doi: 10.1128/jb.92.2.518-520.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Signer E. R. Interaction of prophages at the att80 site with the chromosome of Escherichia coli. J Mol Biol. 1966 Jan;15(1):243–255. doi: 10.1016/s0022-2836(66)80224-3. [DOI] [PubMed] [Google Scholar]
  48. Skalka A. Nucleotide distribution and functional orientation in the deoxyribonucleic acid of phage phi 80. J Virol. 1969 Feb;3(2):150–156. doi: 10.1128/jvi.3.2.150-156.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. TAYLOR A. L., THOMAN M. S. THE GENETIC MAP OF ESCHERICHIA COLI K-12. Genetics. 1964 Oct;50:659–677. doi: 10.1093/genetics/50.4.659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Taylor A. L., Trotter C. D. Linkage map of Escherichia coli strain K-12. Bacteriol Rev. 1972 Dec;36(4):504–524. doi: 10.1128/br.36.4.504-524.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Urm E., Yang H., Zubay G., Kelker N., Maas W. In vitro repression of n- -acetyl-L-ornithinase synthesis in Escherichia coli. Mol Gen Genet. 1973;121(1):1–7. doi: 10.1007/BF00353688. [DOI] [PubMed] [Google Scholar]
  52. Walker J. R., Ussery C. L., Allen J. S. Bacterial cell division regulation: lysogenization of conditional cell division lon - mutants of Escherichia coli by bacteriophage. J Bacteriol. 1973 Mar;113(3):1326–1332. doi: 10.1128/jb.113.3.1326-1332.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wetekam W., Staack K., Ehring R. DNA-dependent in vitro synthesis of enzymes of the galactose operon of Escherichia coli. Mol Gen Genet. 1971;112(1):14–27. doi: 10.1007/BF00266928. [DOI] [PubMed] [Google Scholar]
  54. Wiesmeyer H. Prophage repression as a model for the study of gene regulation. I. Titration of the lambda repressor. J Bacteriol. 1966 Jan;91(1):89–94. doi: 10.1128/jb.91.1.89-94.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Willard M., Echols H. Role of bacteriophage DNA replication in lambda-dg escape synthesis. J Mol Biol. 1968 Feb 28;32(1):37–46. doi: 10.1016/0022-2836(68)90143-5. [DOI] [PubMed] [Google Scholar]
  56. Yamagishi H., Nakamura K., Ozeki H. Cohesion occurring between DNA molecules of temperate phages phi 80 and lambda or phi 81. Biochem Biophys Res Commun. 1965 Sep 22;20(6):727–732. doi: 10.1016/0006-291x(65)90077-x. [DOI] [PubMed] [Google Scholar]
  57. Yamamoto K. R., Alberts B. M., Benzinger R., Lawhorne L., Treiber G. Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification. Virology. 1970 Mar;40(3):734–744. doi: 10.1016/0042-6822(70)90218-7. [DOI] [PubMed] [Google Scholar]
  58. Yarmolinsky M. B., Wiesmeyer H. REGULATION BY COLIPHAGE LAMBDA OF THE EXPRESSION OF THE CAPACITY TO SYNTHESIZE A SEQUENCE OF HOST ENZYMES. Proc Natl Acad Sci U S A. 1960 Dec;46(12):1626–1645. doi: 10.1073/pnas.46.12.1626. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES