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. 1971 May;68(5):1002–1005. doi: 10.1073/pnas.68.5.1002

Enzymatic Characterization of a Mutant of Escherichia coli with an Altered DNA Ligase

Paul Modrich 1, I R Lehman 1
PMCID: PMC389100  PMID: 4995816

Abstract

A temperature-sensitive, radiation-sensitive mutant of Escherichia coli has been assayed for DNA ligase activity in vitro. The strain contains a markedly reduced amount of DNA-joining activity, which is thermolabile. The formation of the ligase-adenylate intermediate is also temperature-sensitive in vitro. Two temperature-resistant revertants of the mutant contain normal amounts of a thermostable ligase. The mutant is killed by growth at 42°C, a temperature at which it displays aberrant DNA synthesis. These results suggest that the ligase is necessary for normal DNA metabolism and viability in this strain.

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Selected References

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

  1. Cozzarelli N. R., Kelly R. B., Kornberg A. A minute circular DNA from Escherichia coli 15. Proc Natl Acad Sci U S A. 1968 Jul;60(3):992–999. doi: 10.1073/pnas.60.3.992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Gellert M., Bullock M. L. DNA ligase mutants of Escherichia coli. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1580–1587. doi: 10.1073/pnas.67.3.1580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Howard-Flanders P. DNA repair. Annu Rev Biochem. 1968;37:175–200. doi: 10.1146/annurev.bi.37.070168.001135. [DOI] [PubMed] [Google Scholar]
  4. Ikeda H., Inuzuka M., Tomizawa J. I. P1-like plasmid in Escherichia coli 15. J Mol Biol. 1970 Jun 14;50(2):457–470. doi: 10.1016/0022-2836(70)90204-4. [DOI] [PubMed] [Google Scholar]
  5. Little J. W., Zimmerman S. B., Oshinsky C. K., Gellert M. Enzymatic joining of DNA strands, II. An enzyme-adenylate intermediate in the dpn-dependent DNA ligase reaction. Proc Natl Acad Sci U S A. 1967 Nov;58(5):2004–2011. doi: 10.1073/pnas.58.5.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Modrich P., Lehman I. R. Enzymatic joining of polynucleotides. IX. A simple and rapid assay of polynucleotide joining (ligase) activity by measurement of circle formation from linear deoxyadenylate-deoxythymidylate copolymer. J Biol Chem. 1970 Jul 25;245(14):3626–3631. [PubMed] [Google Scholar]
  7. Okazaki R., Okazaki T., Sakabe K., Sugimoto K., Sugino A. Mechanism of DNA chain growth. I. Possible discontinuity and unusual secondary structure of newly synthesized chains. Proc Natl Acad Sci U S A. 1968 Feb;59(2):598–605. doi: 10.1073/pnas.59.2.598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Olivera B. M., Hall Z. W., Anraku Y., Chien J. R., Lehman I. R. On the mechanism of the polynucleotide joining reaction. Cold Spring Harb Symp Quant Biol. 1968;33:27–34. doi: 10.1101/sqb.1968.033.01.008. [DOI] [PubMed] [Google Scholar]
  9. Olivera B. M., Lehman I. R. Linkage of polynucleotides through phosphodiester bonds by an enzyme from Escherichia coli. Proc Natl Acad Sci U S A. 1967 May;57(5):1426–1433. doi: 10.1073/pnas.57.5.1426. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Pauling C., Hamm L. Properties of a temperature-sensitive radiation-sensitive mutant of Escherichia coli. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1495–1502. doi: 10.1073/pnas.60.4.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pauling C., Hamm L. Properties of a temperature-sensitive, radiation-sensitive mutant of Escherichia coli. II. DNA replication. Proc Natl Acad Sci U S A. 1969 Dec;64(4):1195–1202. doi: 10.1073/pnas.64.4.1195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Richardson C. C., Masamune Y., Live T. R., Jacquemin-Sablon A., Weiss B., Fareed G. C. Studies on the joining of DNA by polynucleotide ligase of phage T4. Cold Spring Harb Symp Quant Biol. 1968;33:151–164. doi: 10.1101/sqb.1968.033.01.019. [DOI] [PubMed] [Google Scholar]
  13. SHUSTER L., KAPLAN N. O., STOLZENBACH F. E. The reaction of pyridine nucleotides with trifluoroacetic acid anhydride. J Biol Chem. 1955 Jul;215(1):195–209. [PubMed] [Google Scholar]
  14. 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]
  15. Symons R. H. A rapid and improved method for the synthesis of 32P-labelled ribonucleoside 5'-monophosphates. Biochem Biophys Res Commun. 1966 Sep 22;24(6):872–876. doi: 10.1016/0006-291x(66)90329-9. [DOI] [PubMed] [Google Scholar]

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