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. 1974 May;71(5):2048–2051. doi: 10.1073/pnas.71.5.2048

A Conditional Lethal Mutant of Escherichia coli K12 Defective in the 5′ → 3′ Exonuclease Associated with DNA Polymerase I

E Bruce Konrad 1, I R Lehman 1
PMCID: PMC388383  PMID: 4600786

Abstract

A mutant strain of E. coli, initially identified by an abnormally high frequency of recombination, has been found to be defective in the 5′ → 3′ exonuclease associated with DNA polymerase I, but not in the polymerase activity. This defect is tolerated at 30°, but is lethal at 43°. Like other polymerase I mutants, the strain is unusually sensitive to methyl methanesulfonate and to ultraviolet irradiation; it is also unable to support the growth of phage λ defective in general recombination, and shows a reduced rate of joining of 10S “Okazaki fragments.” These results demonstrate that a functional DNA polymerase I is essential for normal growth and viability in E. coli K12.

Keywords: DNA synthesis, UV sensitivity, genetic recombination

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Selected References

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

  1. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bazill G. W., Hall R., Gross J. D. DNA synthesis in lysates of RecB- and Rec+ E. coli cells. Nat New Biol. 1971 Oct 27;233(43):281–283. doi: 10.1038/newbio233281a0. [DOI] [PubMed] [Google Scholar]
  3. Beckwith J. R., Signer E. R. Transposition of the lac region of Escherichia coli. I. Inversion of the lac operon and transduction of lac by phi80. J Mol Biol. 1966 Aug;19(2):254–265. doi: 10.1016/s0022-2836(66)80003-7. [DOI] [PubMed] [Google Scholar]
  4. Brutlag D., Atkinson M. R., Setlow P., Kornberg A. An active fragment of DNA polymerase produced by proteolytic cleavage. Biochem Biophys Res Commun. 1969 Dec 4;37(6):982–989. doi: 10.1016/0006-291x(69)90228-9. [DOI] [PubMed] [Google Scholar]
  5. De Lucia P., Cairns J. Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature. 1969 Dec 20;224(5225):1164–1166. doi: 10.1038/2241164a0. [DOI] [PubMed] [Google Scholar]
  6. Glickman B. W., van Sluis C. A., Heijneker H. L., Rörsch A. A mutant of Escherichia coli K12 deficient in the 5'-3' exonucleolytic activity of DNA polymerase I. I. General characterization. Mol Gen Genet. 1973 Jul 31;124(1):69–82. doi: 10.1007/BF00267166. [DOI] [PubMed] [Google Scholar]
  7. Heijneker H. L., Ellens D. J., Tjeerde R. H., Glickman B. W., van Dorp B., Pouwels P. H. A mutant of Escherichia coli K12 deficient in the 5'-3' exonucleolytic activity of DNA polymerase I. II. Purification and properties of the mutant enzyme. Mol Gen Genet. 1973 Jul 31;124(1):83–96. doi: 10.1007/BF00267167. [DOI] [PubMed] [Google Scholar]
  8. Hirose S., Okazaki R., Tamanoi F. Mechanism of DNA chain growth. XI. Structure of RNA-linked DNA fragments of Escherichia coli. J Mol Biol. 1973 Jul 15;77(4):501–517. doi: 10.1016/0022-2836(73)90219-2. [DOI] [PubMed] [Google Scholar]
  9. Kato T., Kondo S. Genetic and molecular characteristics of X-ray-sensitive mutants of Escherichia coli defective in repair synthesis. J Bacteriol. 1970 Nov;104(2):871–881. doi: 10.1128/jb.104.2.871-881.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Klenow H., Henningsen I. Selective elimination of the exonuclease activity of the deoxyribonucleic acid polymerase from Escherichia coli B by limited proteolysis. Proc Natl Acad Sci U S A. 1970 Jan;65(1):168–175. doi: 10.1073/pnas.65.1.168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Konrad E. B., Modrich P., Lehman I. R. Genetic and enzymatic characterization of a conditional lethal mutant of Escherichia coli K12 with a temperature-sensitive DNA ligase. J Mol Biol. 1973 Jul 15;77(4):519–529. doi: 10.1016/0022-2836(73)90220-9. [DOI] [PubMed] [Google Scholar]
  12. Kuempel P. L., Veomett G. E. A possible function of DNA polymerase in chromosome replication. Biochem Biophys Res Commun. 1970 Nov 25;41(4):973–980. doi: 10.1016/0006-291x(70)90180-4. [DOI] [PubMed] [Google Scholar]
  13. Lehman I. R., Chien J. R. Persistence of deoxyribonucleic acid polymerase I and its 5'--3' exonuclease activity in PolA mutants of Escherichia coli K12. J Biol Chem. 1973 Nov 25;248(22):7717–7723. [PubMed] [Google Scholar]
  14. Low K. B. Escherichia coli K-12 F-prime factors, old and new. Bacteriol Rev. 1972 Dec;36(4):587–607. doi: 10.1128/br.36.4.587-607.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Monk M., Kinross J. Conditional lethality of recA and recB derivatives of a strain of Escherichia coli K-12 with a temperature-sensitive deoxyribonucleic acid polymerase I. J Bacteriol. 1972 Mar;109(3):971–978. doi: 10.1128/jb.109.3.971-978.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Okazaki R., Arisawa M., Sugino A. Slow joining of newly replicated DNA chains in DNA polymerase I-deficient Escherichia coli mutants. Proc Natl Acad Sci U S A. 1971 Dec;68(12):2954–2957. doi: 10.1073/pnas.68.12.2954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. STEINBERG C. M., EDGAR R. S. A critical test of a current theory of genetic recombination in bacteriophage. Genetics. 1962 Feb;47:187–208. doi: 10.1093/genetics/47.2.187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]

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