Abstract
DNA damage generated by oxygen radicals includes base-free apurinic/apyrimidinic (AP) sites and strand breaks that bear deoxyribose fragments. The yeast Saccharomyces cerevisiae repairs such DNA lesions by using a single major enzyme. We have cloned the yeast structural gene (APN1) encoding this AP endonuclease/3'-repair diesterase by immunological screening of a yeast genomic DNA expression library in lambda gt11. Gene disruption experiments confirm that the Apn1 protein accounts for greater than or equal to 97% of both AP endonuclease and DNA 3'-repair diesterase activities in yeast cell-free extracts. The DNA and predicted amino acid sequences for the APN1 gene are homologous to those for the nfo gene encoding DNA endonuclease IV of Escherichia coli. This conservation of structure between a eukaryotic enzyme and its prokaryotic counterpart underscores the fundamental nature of their roles in DNA repair.
MeSH Terms
Amino Acid Sequence
Base Sequence
Blotting, Western
Cloning, Molecular
DNA Repair
DNA-(Apurinic or Apyrimidinic Site) Lyase
Deoxyribonuclease IV (Phage T4-Induced)
Endodeoxyribonucleases/genetics
Escherichia coli/enzymology,genetics
Escherichia coli Proteins
Genes, Fungal
Molecular Sequence Data
Restriction Mapping
Saccharomyces cerevisiae/enzymology,genetics
Chemicals
Escherichia coli Proteins
Endodeoxyribonucleases
Deoxyribonuclease IV (Phage T4-Induced)
endonuclease IV, E coli
DNA-(Apurinic or Apyrimidinic Site) Lyase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Popoff S C
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
Spira A I
Johnson A W
Demple B
References (25)
25 references, click to expand
-
Endonuclease IV (nfo) mutant of Escherichia coli.
J Bacteriol. 1986 Dec;168(3):1120-7
PMID: 2430946
-
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
Anal Biochem. 1976 May 7;72:248-54
PMID: 942051
-
Chemical changes induced in DNA by ionizing radiation.
Prog Nucleic Acid Res Mol Biol. 1985;32:115-54
PMID: 3003798
-
Yeast DNA 3'-repair diesterase is the major cellular apurinic/apyrimidinic endonuclease: substrate specificity and kinetics.
J Biol Chem. 1988 Dec 5;263(34):18017-22
PMID: 3056936
-
Yeast DNA diesterase for 3'-fragments of deoxyribose: purification and physical properties of a repair enzyme for oxidative DNA damage.
J Biol Chem. 1988 Dec 5;263(34):18009-16
PMID: 3056935
-
Nucleotide sequence of the xth gene of Escherichia coli K-12.
J Bacteriol. 1988 Oct;170(10):4542-7
PMID: 3049539
-
DNA damage and oxygen radical toxicity.
Science. 1988 Jun 3;240(4857):1302-9
PMID: 3287616
-
Purification and properties of the major nuclease from mitochondria of Saccharomyces cerevisiae.
J Biol Chem. 1988 Jun 5;263(16):7691-702
PMID: 3286639
-
Multiple DNA repair activities for 3'-deoxyribose fragments in Escherichia coli.
Nucleic Acids Res. 1989 Jan 25;17(2):587-600
PMID: 2464796
-
Nucleotide sequence of the nfo gene of Escherichia coli K-12.
J Bacteriol. 1988 Nov;170(11):5141-5
PMID: 2460435
-
AP endonucleases and DNA glycosylases that recognize oxidative DNA damage.
Environ Mol Mutagen. 1988;12(4):431-77
PMID: 2461302
-
Homogeneous Escherichia coli endonuclease IV. Characterization of an enzyme that recognizes oxidative damage in DNA.
J Biol Chem. 1988 Jun 15;263(17):8066-71
PMID: 2453510
-
Phosphodiesterases involved in DNA repair.
Adv Enzymol Relat Areas Mol Biol. 1987;60:1-34
PMID: 2444076
-
Endonuclease IV of Escherichia coli is induced by paraquat.
Proc Natl Acad Sci U S A. 1987 May;84(10):3189-93
PMID: 2437576
-
Exonuclease III and endonuclease IV remove 3' blocks from DNA synthesis primers in H2O2-damaged Escherichia coli.
Proc Natl Acad Sci U S A. 1986 Oct;83(20):7731-5
PMID: 2429316
-
Endonuclease-resistant apyrimidinic sites formed by neocarzinostatin at cytosine residues in DNA: evidence for a possible role in mutagenesis.
Proc Natl Acad Sci U S A. 1985 May;82(10):3182-6
PMID: 2582408
-
Genetic map of Saccharomyces cerevisiae, edition 10.
Yeast. 1989 Sep-Oct;5(5):321-403
PMID: 2678811
-
Yeast/E. coli shuttle vectors with multiple unique restriction sites.
Yeast. 1986 Sep;2(3):163-7
PMID: 3333305
-
A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains.
Genetics. 1987 Aug;116(4):541-5
PMID: 3305158
-
DNA repair enzymes.
Annu Rev Biochem. 1982;51:61-87
PMID: 6287922
-
Yeast RNA polymerase II genes: isolation with antibody probes.
Science. 1983 Nov 18;222(4625):778-82
PMID: 6356359
-
Transformation of intact yeast cells treated with alkali cations.
J Bacteriol. 1983 Jan;153(1):163-8
PMID: 6336730
-
Escherichia coli xth mutants are hypersensitive to hydrogen peroxide.
J Bacteriol. 1983 Feb;153(2):1079-82
PMID: 6337115
-
A comprehensive set of sequence analysis programs for the VAX.
Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387-95
PMID: 6546423
-
Oxidants and human disease: some new concepts.
FASEB J. 1987 Nov;1(5):358-64
PMID: 2824268