Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1979 Dec;140(3):825–834. doi: 10.1128/jb.140.3.825-834.1979

Conditionally lethal amber mutations in the dnaA region of the Escherichia coli chromosome that affect chromosome replication.

M Kimura, T Miki, S Hiraga, T Nagata, T Yura
PMCID: PMC216721  PMID: 160413

Abstract

Three amber mutations, dna-801, dna-803, and dna-806, were isolated by localized mutagenesis of the dnaA-oriC region of the chromosome from an Escherichia coli strain carrying temperature-sensitive amber suppressors. When the mutations were not suppressed at 42 degrees C, the cells did not grow and DNA synthesis was arrested. They were very closely linked to each other and to the dnaA46 mutation. The mutant phenotype of each strain was converted to the wild type by infecting the mutants with specialized transducing phase lambda i21 dnaA-2 but not with lambda i21 tna. Derivatives of lambda i21 dnaA-2, each of which carried the amber mutation dna-801 dna-803, or dna-806, converted the dnaA mutant phenotype to Dna+ but did not convert rhe amber mutants to the wild-type phenotype. E. coli uvrB cells were irradiated with ultraviolet light and infected with each of these phage strains. An analysis of proteins synthesized in the cells revealed that two proteins with molecular weights of 50,000 and 43,000 were specified by lambda i21 dnaA-2 but not by lambda i21 tna. When the ultraviolet-irradiated cells did not carry an amber suppressor, the derivative phage with the amber mutation invariably failed to produce the 43,000-dalton protein, but when the host cell carried supF (tyrT), the protein was produced. The 50,000-dalton protein was unaffected.

Full text

PDF
825

Images in this article

830Image
on p.830
831Image
on p.831
832Image
on p.832

Selected References

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

  1. Abe M., Tomizawa J. Chromosome replication in Escherichia coli K12 mutant affected in the process of DNA initiation. Genetics. 1971 Sep;69(1):1–15. doi: 10.1093/genetics/69.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andoh T., Ozeki H. Suppressor gene Su3+ of E. coli, a structural gene for tyrosine TRNA. Proc Natl Acad Sci U S A. 1968 Mar;59(3):792–799. doi: 10.1073/pnas.59.3.792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baas P. D., Jansz H. S. Bacteriophage phiX174 DNA synthesis in a replication-deficient host: determination of the origin of phiX DNA replication. J Mol Biol. 1976 Apr 15;102(3):633–656. doi: 10.1016/0022-2836(76)90339-9. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Berger I., Geyl D., Böck A., Stöffler G., Wittmann H. G. Localized mutagenesis of the aroE-strA section of the Escherichia coli chromosome coding for ribosomal proteins. Mol Gen Genet. 1975 Dec 1;141(3):207–211. doi: 10.1007/BF00341800. [DOI] [PubMed] [Google Scholar]
  6. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  7. Borck K., Beggs J. D., Brammar W. J., Hopkins A. S., Murray N. E. The construction in vitro of transducing derivatives of phage lambda. Mol Gen Genet. 1976 Jul 23;146(2):199–207. doi: 10.1007/BF00268089. [DOI] [PubMed] [Google Scholar]
  8. Butlin J. D., Cox G. B., Gibson F. Oxidative phosphorylation in Escherichia coli K12. Mutations affecting magnesium ion- or calcium ion-stimulated adenosine triphosphatase. Biochem J. 1971 Aug;124(1):75–81. doi: 10.1042/bj1240075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Carl P. L. Escherichia coli mutants with temperature-sensitive synthesis of DNA. Mol Gen Genet. 1970;109(2):107–122. doi: 10.1007/BF00269647. [DOI] [PubMed] [Google Scholar]
  10. Furth M. E., Blattner F. R., McLeester C., Dove W. F. Genetic structure of the replication origin of bacteriophage lambda. Science. 1977 Dec 9;198(4321):1046–1051. doi: 10.1126/science.929186. [DOI] [PubMed] [Google Scholar]
  11. Hiraga S. Novel F prime factors able to replicate in Escherichia coli Hfr strains. Proc Natl Acad Sci U S A. 1976 Jan;73(1):198–202. doi: 10.1073/pnas.73.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hirota Y., Mordoh J., Jacob F. On the process of cellular division in Escherichia coli. 3. Thermosensitive mutants of Escherichia coli altered in the process of DNA initiation. J Mol Biol. 1970 Nov 14;53(3):369–387. doi: 10.1016/0022-2836(70)90072-0. [DOI] [PubMed] [Google Scholar]
  13. Hong J. S., Ames B. N. Localized mutagenesis of any specific small region of the bacterial chromosome. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3158–3162. doi: 10.1073/pnas.68.12.3158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Horiuchi T., Nagata T. Mutations affecting growth of the Escherichia coli cell under a condition of DNA polymerase I-deficiency. Mol Gen Genet. 1973;123(1):89–110. doi: 10.1007/BF00282992. [DOI] [PubMed] [Google Scholar]
  15. Ikeda H., Tomizawa J. I. Transducing fragments in generalized transduction by phage P1. I. Molecular origin of the fragments. J Mol Biol. 1965 Nov;14(1):85–109. doi: 10.1016/s0022-2836(65)80232-7. [DOI] [PubMed] [Google Scholar]
  16. Joh K., Hiraga S. Genetic mapping of the chromosomal replication origin of Salmonella typhimurium. J Bacteriol. 1979 May;138(2):297–304. doi: 10.1128/jb.138.2.297-304.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kanazawa H., Miki T., Tamura F., Yura T., Futai M. Specialized transducing phage lambda carrying the genes for coupling factor of oxidative phosphorylation of Escherichia coli: increased synthesis of coupling factor on induction of prophage lambda asn. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1126–1130. doi: 10.1073/pnas.76.3.1126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Karamata D., Gross J. D. Isolation and genetic analysis of temperature-sensitive mutants of B. subtilis defective in DNA synthesis. Mol Gen Genet. 1970;108(3):277–287. doi: 10.1007/BF00283358. [DOI] [PubMed] [Google Scholar]
  19. Kellenberger-Gujer G., Podhajska A. J., Caro L. A cold sensitive dnaA mutant of E. coli which overinitiates chromosome replication at low temperature. Mol Gen Genet. 1978 Jun 1;162(1):9–16. doi: 10.1007/BF00333845. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Miki T., Hiraga S., Nagata T., Yura T. Bacteriophage lambda carrying the Escherichia coli chromosomal region of the replication origin. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5099–5103. doi: 10.1073/pnas.75.10.5099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Miki T., Kimura M., Hiraga S., Nagata T., Yura T. Cloning and physical mapping of the dnaA region of the Escherichia coli chromosome. J Bacteriol. 1979 Dec;140(3):817–824. doi: 10.1128/jb.140.3.817-824.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Murgola E. J., Yanofsky C. Structural interactions between amino acid residues at positions 22 and 211 in the tryptophan synthetase alpha chain of Escherichia coli. J Bacteriol. 1974 Feb;117(2):444–448. doi: 10.1128/jb.117.2.444-448.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Murray K., Murray N. E. Phage lambda receptor chromosomes for DNA fragments made with restriction endonuclease III of Haemophilus influenzae and restriction endonuclease I of Escherichia coli. J Mol Biol. 1975 Nov 5;98(3):551–564. doi: 10.1016/s0022-2836(75)80086-6. [DOI] [PubMed] [Google Scholar]
  25. Nagata T., Horiuchi T. An amber dna mutant of Escherichia coli K12 affecting DNA ligase. J Mol Biol. 1974 Aug 5;87(2):369–373. doi: 10.1016/0022-2836(74)90156-9. [DOI] [PubMed] [Google Scholar]
  26. Nagata T., Horiuchi T. Isolation and characterization of a temperature-sensitive amber suppressor mutant of Escherichia coli K12. Mol Gen Genet. 1973;123(1):77–88. doi: 10.1007/BF00282991. [DOI] [PubMed] [Google Scholar]
  27. Schaefler S., Maas W. K. Inducible system for the utilization of beta-glucosides in Escherichia coli. II. Description of mutant types and genetic analysis. J Bacteriol. 1967 Jan;93(1):264–272. doi: 10.1128/jb.93.1.264-272.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wechsler J. A., Gross J. D. Escherichia coli mutants temperature-sensitive for DNA synthesis. Mol Gen Genet. 1971;113(3):273–284. doi: 10.1007/BF00339547. [DOI] [PubMed] [Google Scholar]
  29. Yamamori T., Ito K., Nakamura Y., Yura T. Transient regulation of protein synthesis in Escherichia coli upon shift-up of growth temperature. J Bacteriol. 1978 Jun;134(3):1133–1140. doi: 10.1128/jb.134.3.1133-1140.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yasuda S., Hirota Y. Cloning and mapping of the replication origin of Escherichia coli. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5458–5462. doi: 10.1073/pnas.74.12.5458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. von Meyenburg K., Hansen F. G., Nielsin L. D., Riise E. Origin of replication, oriC, or the Escherichia coli chromosome on specialized transducing phages lambda asn. Mol Gen Genet. 1978 Apr 17;160(3):287–295. doi: 10.1007/BF00332972. [DOI] [PubMed] [Google Scholar]

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

RESOURCES