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Proc Natl Acad Sci U S A. 1985 Jan; 82(2): 474–478.
PMCID: PMC397061

Endonuclease III (nth) mutants of Escherichia coli.


Two strains that overproduce endonuclease III were found in a colony bank containing hybrid ColE1-Escherichia coli plasmids. The enzyme was identified in crude extracts by the degradation of partially depyrimidinated DNA in the presence of EDTA, by its sedimentation velocity, and by its associated thymine glycol-DNA glycosylase activity. An insertion mutation was produced by cloning the kanamycin-resistance gene of Tn5 into the plasmid copy of the nth gene. The mutation was then transferred to the chromosome in the following steps: (i) selection for chromosomal integration of the plasmid at 42 degrees C in a temperature-sensitive polA strain, (ii) curing via temperature shifts, and (iii) phage P1-mediated transduction of a new host. The insertion mutant, as well as a separately isolated deletion mutant, had no measurable glycosylase activity for DNA containing thymine glycol. Although such residues are common lesions in oxidized or irradiated DNA, the mutants were not unusually sensitive to H2O2 or gamma-rays. The insertion mutation had a mutator effect (4- to 22-fold enhancement) on one tested allele.

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  • Burton K, Riley WT. Selective degradation of thymidine and thymine deoxynucleotides. Biochem J. 1966 Jan;98(1):70–77. [PMC free article] [PubMed]
  • Frenkel K, Goldstein MS, Duker NJ, Teebor GW. Identification of the cis-thymine glycol moiety in oxidized deoxyribonucleic acid. Biochemistry. 1981 Feb 17;20(4):750–754. [PubMed]
  • Hariharan PV, Achey PM, Cerutti PA. Biological effect of thymine ring saturation in coliphage phiX174-DNA. Radiat Res. 1977 Feb;69(2):375–378. [PubMed]
  • Demple B, Linn S. DNA N-glycosylases and UV repair. Nature. 1980 Sep 18;287(5779):203–208. [PubMed]
  • Bruyninckx WJ, Mason HS, Morse SA. Are physiological oxygen concentrations mutagenic? Nature. 1978 Aug 10;274(5671):606–607. [PubMed]
  • Teoule R, Bert C, Bonicel A. Thymine fragment damage retained in the DNA polynucleotide chain after gamma irradiation in aerated solutions. II. Radiat Res. 1977 Nov;72(2):190–200. [PubMed]
  • Gates FT, Linn S. Endonuclease from Escherichia coli that acts specifically upon duplex DNA damaged by ultraviolet light, osmium tetroxide, acid, or x-rays. J Biol Chem. 1977 May 10;252(9):2802–2807. [PubMed]
  • Warner HR, Demple BF, Deutsch WA, Kane CM, Linn S. Apurinic/apyrimidinic endonucleases in repair of pyrimidine dimers and other lesions in DNA. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4602–4606. [PMC free article] [PubMed]
  • Katcher HL, Wallace SS. Characterization of the Escherichia coli X-ray endonuclease, endonuclease III. Biochemistry. 1983 Aug 16;22(17):4071–4081. [PubMed]
  • Breimer LH, Lindahl T. DNA glycosylase activities for thymine residues damaged by ring saturation, fragmentation, or ring contraction are functions of endonuclease III in Escherichia coli. J Biol Chem. 1984 May 10;259(9):5543–5548. [PubMed]
  • Lindahl T. DNA repair enzymes. Annu Rev Biochem. 1982;51:61–87. [PubMed]
  • Weiss B, Milcarek C. Mass screening for mutants with altered DNases by microassay techniques. Methods Enzymol. 1974;29:180–193. [PubMed]
  • Warner HR, Duncan BK. In vivo synthesis and properties of uracil-containing DNA. Nature. 1978 Mar 2;272(5648):32–34. [PubMed]
  • Lindahl T, Ljungquist S, Siegert W, Nyberg B, Sperens B. DNA N-glycosidases: properties of uracil-DNA glycosidase from Escherichia coli. J Biol Chem. 1977 May 25;252(10):3286–3294. [PubMed]
  • Paquette Y, Crine P, Verly WG. Properties of the endonuclease for depurinated DNA from Escherichia coli. Can J Biochem. 1972 Nov;50(11):1199–1209. [PubMed]
  • Birnboim HC, Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. [PMC free article] [PubMed]
  • Milcarek C, Weiss B. Mutants of Escherichia coli with altered deoxyribonucleases. I. Isolation and characterization of mutants for exonuclease 3. J Mol Biol. 1972 Jul 21;68(2):303–318. [PubMed]
  • Rhoades M, MacHattie LA, Thomas CA., Jr The P22 bacteriophage DNA molecule. I. The mature form. J Mol Biol. 1968 Oct 14;37(1):21–40. [PubMed]
  • Hochhauser SJ, Weiss B. Escherichia coli mutants deficient in deoxyuridine triphosphatase. J Bacteriol. 1978 Apr;134(1):157–166. [PMC free article] [PubMed]
  • Clarke L, Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. [PubMed]
  • Ljungquist S. A new endonuclease from Escherichia coli acting at apurinic sites in DNA. J Biol Chem. 1977 May 10;252(9):2808–2814. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • MARTIN RG, AMES BN. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed]
  • Iida S, Hayatsu H. The permanganate oxidation of thymine. Biochim Biophys Acta. 1970 Jul 16;213(1):1–13. [PubMed]
  • Jorgensen RA, Rothstein SJ, Reznikoff WS. A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. Mol Gen Genet. 1979;177(1):65–72. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Yamaguchi K, Tomizawa J. Establishment of Escherichia coli cells with an integrated high copy number plasmid. Mol Gen Genet. 1980;178(3):525–533. [PubMed]
  • Jochimsen B, Nygaard P, Vestergaard T. Location on the chromosome of Escherichia coli of genes governing purine metabolism. Adenosine deaminase (add), guanosine kinase (gsk) and hypoxanthine phosphoribosyltransferase (hpt). Mol Gen Genet. 1975 Dec 30;143(1):85–91. [PubMed]
  • Fouts KE, Barbour SD. Transductional mapping of ksgB and a new Tn5-induced kasugamycin resistance gene, ksgD, in Escherichia coli K-12. J Bacteriol. 1981 Feb;145(2):914–919. [PMC free article] [PubMed]
  • Radman M. An endonuclease from Escherichia coli that introduces single polynucleotide chain scissions in ultraviolet-irradiated DNA. J Biol Chem. 1976 Mar 10;251(5):1438–1445. [PubMed]
  • Bachmann BJ. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. [PMC free article] [PubMed]
  • Demple B, Halbrook J, Linn S. Escherichia coli xth mutants are hypersensitive to hydrogen peroxide. J Bacteriol. 1983 Feb;153(2):1079–1082. [PMC free article] [PubMed]
  • Walker GC. Plasmid (pKM101)-mediated enhancement of repair and mutagenesis: dependence on chromosomal genes in Escherichia coli K-12. Mol Gen Genet. 1977 Mar 28;152(1):93–103. [PubMed]
  • Duncan BK, Weiss B. Specific mutator effects of ung (uracil-DNA glycosylase) mutations in Escherichia coli. J Bacteriol. 1982 Aug;151(2):750–755. [PMC free article] [PubMed]
  • Walker GC. Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev. 1984 Mar;48(1):60–93. [PMC free article] [PubMed]

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