Genetic Analysis of Components Involved in Vitamin B12 Uptake in Escherichia coli (original) (raw)

Abstract

The products of three genes are involved in cyanocobalamin (B12) uptake in Escherichia coli. btuB (formerly bfe), located at min 88 on the Escherichia coli linkage map, codes for a protein component of the outer membrane which serves as receptor for B12, the E colicins, and bacteriophage BF23. Four phenotypic classes of mutants varying in response to these agents were found to carry mutations that, based on complementation and reversion analyses, reside in the single btuB cistron. In one mutant class, ligand binding to the receptor appeared to be normal, but subsequent B12 uptake was defective. The level of receptor and rate of uptake were responsive to btuB gene dosage. Previous studies showed that the tonB product was necessary for energy-dependent B12 uptake but not for its binding. Other than those in tonB, no mutations that conferred insensitivity to group B colicins affected B12 utilization. The requirement for the btuB and tonB products could be bypassed by elevated levels of B12 (>1 μM) or by mutations compromising the integrity of the outer membrane as a permeability barrier. Utilization of elevated B12 concentrations in strains lacking the btuB-tonB uptake system was dependent on the function of the btuC product. This gene was located at 37.7 min on the linkage map, with the order pps-btuC-pheS. Strains altered in btuC but with an intact btuB-tonB system were only slightly impaired in B12 utilization, being defective in its accumulation. This defect was manifested as inability to retain B12, such that intracellular label was almost completely lost by exchange or efflux. It is proposed that btuC encodes a transport system for B12 in the periplasm.

796

Selected References

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

  1. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bassford P. J., Jr, Bradbeer C., Kadner R. J., Schnaitman C. A. Transport of vitamin B12 in tonB mutants of Escherichia coli. J Bacteriol. 1976 Oct;128(1):242–247. doi: 10.1128/jb.128.1.242-247.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradbeer C., Woodrow M. L., Khalifah L. I. Transport of vitamin B12 in Escherichia coli: common receptor system for vitamin B12 and bacteriophage BF23 on the outer membrane of the cell envelope. J Bacteriol. 1976 Mar;125(3):1032–1039. doi: 10.1128/jb.125.3.1032-1039.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradbeer C., Woodrow M. L. Transport of vitamin B12 in Escherichia coli: energy dependence. J Bacteriol. 1976 Oct;128(1):99–104. doi: 10.1128/jb.128.1.99-104.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buxton R. S. Genetic analysis of Escherichia coli K12 mutants resistant to bacteriophage BF23 and the E-group colicins. Mol Gen Genet. 1971;113(2):154–156. doi: 10.1007/BF00333188. [DOI] [PubMed] [Google Scholar]
  6. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Datta D. B., Krämer C., Henning U. Diploidy for a structural gene specifying a major protein of the outer cell envelope membrane from Escherichia coli K-12. J Bacteriol. 1976 Dec;128(3):834–841. doi: 10.1128/jb.128.3.834-841.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davies J. K., Reeves P. Genetics of resistance to colicins in Escherichia coli K-12: cross-resistance among colicins of group B. J Bacteriol. 1975 Jul;123(1):96–101. doi: 10.1128/jb.123.1.96-101.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Di Girolamo P. M., Bradbeer C. Transport of vitamin B 12 in Escherichia coli. J Bacteriol. 1971 Jun;106(3):745–750. doi: 10.1128/jb.106.3.745-750.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Di Girolamo P. M., Kadner R. J., Bradbeer C. Isolation of vitamin B 12 transport mutants of Escherichia coli. J Bacteriol. 1971 Jun;106(3):751–757. doi: 10.1128/jb.106.3.751-757.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Di Masi D. R., White J. C., Schnaitman C. A., Bradbeer C. Transport of vitamin B12 in Escherichia coli: common receptor sites for vitamin B12 and the E colicins on the outer membrane of the cell envelope. J Bacteriol. 1973 Aug;115(2):506–513. doi: 10.1128/jb.115.2.506-513.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ennis H. L. Mutants of Escherichia coli sensitive to antibiotics. J Bacteriol. 1971 Aug;107(2):486–490. doi: 10.1128/jb.107.2.486-490.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Frost G. E., Rosenberg H. Relationship between the tonB locus and iron transport in Escherichia coli. J Bacteriol. 1975 Nov;124(2):704–712. doi: 10.1128/jb.124.2.704-712.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hantke K., Braun V. Membrane receptor dependent iron transport in Escherichia coli. FEBS Lett. 1975 Jan 1;49(3):301–305. doi: 10.1016/0014-5793(75)80771-x. [DOI] [PubMed] [Google Scholar]
  15. Jasper P., Whitney E., Silver S. Genetic locus determining resistance to phage BF23 and colicins E 1 , E 2 and E 3 in Escherichia coli. Genet Res. 1972 Jun;19(3):305–312. doi: 10.1017/s0016672300014555. [DOI] [PubMed] [Google Scholar]
  16. Kadner R. J., Bassford P. J., Jr Relation of cell growth and colicin tolerance to vitamin B12 uptake in Escherichia coli. J Bacteriol. 1977 Jan;129(1):254–264. doi: 10.1128/jb.129.1.254-264.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kadner R. J., Liggins G. L. Transport of vitamin B12 in Escherichia coli: genetic studies. J Bacteriol. 1973 Aug;115(2):514–521. doi: 10.1128/jb.115.2.514-521.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kadner R. J., Winkler H. H. Isolation and characterization of mutations affecting the transport of hexose phosphates in Escherichia coli. J Bacteriol. 1973 Feb;113(2):895–900. doi: 10.1128/jb.113.2.895-900.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Low B. Formation of merodiploids in matings with a class of Rec- recipient strains of Escherichia coli K12. Proc Natl Acad Sci U S A. 1968 May;60(1):160–167. doi: 10.1073/pnas.60.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oeschger M. P., Berlyn M. K. A simple procedure for localized mutagenesis using nitrosoguanidine. Mol Gen Genet. 1974;134(1):77–83. doi: 10.1007/BF00332814. [DOI] [PubMed] [Google Scholar]
  21. Pugsley A. P., Reeves P. Iron uptake in colicin B-resistant mutants of Escherichia coli K-12. J Bacteriol. 1976 Jun;126(3):1052–1062. doi: 10.1128/jb.126.3.1052-1062.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Radke K. L., Siegel E. C. Mutation preventing capsular polysaccharide synthesis in Escherichia coli K-12 and its effect on bacteriophage resistance. J Bacteriol. 1971 May;106(2):432–437. doi: 10.1128/jb.106.2.432-437.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sabet S. F., Schnaitman C. A. Purification and properties of the colicin E3 receptor of Escherichia coli. J Biol Chem. 1973 Mar 10;248(5):1797–1806. [PubMed] [Google Scholar]
  24. Tamaki S., Sato T., Matsuhashi M. Role of lipopolysaccharides in antibiotic resistance and bacteriophage adsorption of Escherichia coli K-12. J Bacteriol. 1971 Mar;105(3):968–975. doi: 10.1128/jb.105.3.968-975.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Taylor R. T., Norrell S. A., Hanna M. L. Uptake of cyanocobalamin by Escherichia coli B: some characteristics and evidence for a binding protein. Arch Biochem Biophys. 1972 Feb;148(2):366–381. doi: 10.1016/0003-9861(72)90154-3. [DOI] [PubMed] [Google Scholar]
  26. Vinopal R. T., Fraenkel D. G. PfkB and pfkC loci of Escherichia coli. J Bacteriol. 1975 Jun;122(3):1153–1161. doi: 10.1128/jb.122.3.1153-1161.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. White B. J., Hochhauser S. J., Cintron N. M., Weiss B. Genetic mapping of xthA, the structural gene for exonuclease III in Escherichia coli K-12. J Bacteriol. 1976 Jun;126(3):1082–1088. doi: 10.1128/jb.126.3.1082-1088.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. White J. C., DiGirolamo P. M., Fu M. L., Preston Y. A., Bradbeer C. Transport of vitamin B 12 in Escherichia coli. Location and properties of the initial B 12 -binding site. J Biol Chem. 1973 Jun 10;248(11):3978–3986. [PubMed] [Google Scholar]
  29. Zipkas D., Riley M. Proposal concerning mechanism of evolution of the genome of Escherichia coli. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1354–1358. doi: 10.1073/pnas.72.4.1354. [DOI] [PMC free article] [PubMed] [Google Scholar]