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. 1996 May 15;24(10):1950–1953. doi: 10.1093/nar/24.10.1950

Apurinic/apyrimidinic (AP) endonuclease from Dictyostelium discoideum: cloning, nucleotide sequence and induction by sublethal levels of DNA damaging agents.

T M Freeland 1, R B Guyer 1, A Z Ling 1, R A Deering 1
PMCID: PMC145883  PMID: 8657579

Abstract

We have cloned an AP endonuclease gene (APEA) from Dictyostelium discoideum, along with 1.8 kb of the 5' flanking region. There are no introns. The sequence predicts a protein of 361 amino acids, showing high homology to the major human/Escherichia coli exonuclease III family of AP endonucleases. There is 47% identity and 64% similarity to the Ape endonuclease of human cells using the C-terminal 257 amino acids of the Dictyostelium protein. The 104 amino acids on the N-terminus show only low homology with other AP endonucleases. Instead, this region shows high homology with the acid-rich regions of proteins associated with chromatin, such as nucleolins and HMG proteins. The gene is transcriptionally activated up to 7-fold after treatment of cells with sublethal levels of DNA damaging agents, including ultraviolet light, MNNG and bleomycin. Induction does not occur following blocking of replication fork polymerases with aphidicolin. It is not eliminated by treatment with kinase or phosphatase inhibitors. Four DNA damage-sensitive mutants all retained the DNA damage-induced up-regulation.

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Selected References

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  1. Barzilay G., Walker L. J., Robson C. N., Hickson I. D. Site-directed mutagenesis of the human DNA repair enzyme HAP1: identification of residues important for AP endonuclease and RNase H activity. Nucleic Acids Res. 1995 May 11;23(9):1544–1550. doi: 10.1093/nar/23.9.1544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bronner C. E., Welker D. L., Deering R. A. Mutations affecting sensitivity of the cellular slime mold Dictyostelium discoideum to DNA-damaging agents. Mutat Res. 1992 Sep;274(3):187–200. doi: 10.1016/0921-8777(92)90065-b. [DOI] [PubMed] [Google Scholar]
  3. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  4. Demple B., Harrison L. Repair of oxidative damage to DNA: enzymology and biology. Annu Rev Biochem. 1994;63:915–948. doi: 10.1146/annurev.bi.63.070194.004411. [DOI] [PubMed] [Google Scholar]
  5. Fornace A. J., Jr, Zmudzka B., Hollander M. C., Wilson S. H. Induction of beta-polymerase mRNA by DNA-damaging agents in Chinese hamster ovary cells. Mol Cell Biol. 1989 Feb;9(2):851–853. doi: 10.1128/mcb.9.2.851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guyer R. B., Deering R. A. Apurinic/apyrimidinic-specific endonuclease activities from Dictyostelium discoideum. Biochim Biophys Acta. 1985 Apr 19;824(4):304–312. doi: 10.1016/0167-4781(85)90036-3. [DOI] [PubMed] [Google Scholar]
  7. Guyer R. B., Nonnemaker J. M., Deering R. A. Uracil-DNA glycosylase activity from Dictyostelium discoideum. Biochim Biophys Acta. 1986 Dec 18;868(4):262–264. doi: 10.1016/0167-4781(86)90063-1. [DOI] [PubMed] [Google Scholar]
  8. Luethy J. D., Holbrook N. J. Activation of the gadd153 promoter by genotoxic agents: a rapid and specific response to DNA damage. Cancer Res. 1992 Jan 1;52(1):5–10. [PubMed] [Google Scholar]
  9. Mauldin S. K., Freeland T. M., Deering R. A. Differential repair of UV damage in a developmentally regulated gene of Dictyostelium discoideum. Mutat Res. 1994 Mar;314(2):187–198. doi: 10.1016/0921-8777(94)90082-5. [DOI] [PubMed] [Google Scholar]
  10. Navas T. A., Zhou Z., Elledge S. J. DNA polymerase epsilon links the DNA replication machinery to the S phase checkpoint. Cell. 1995 Jan 13;80(1):29–39. doi: 10.1016/0092-8674(95)90448-4. [DOI] [PubMed] [Google Scholar]
  11. Nellen W., Datta S., Reymond C., Sivertsen A., Mann S., Crowley T., Firtel R. A. Molecular biology in Dictyostelium: tools and applications. Methods Cell Biol. 1987;28:67–100. doi: 10.1016/s0091-679x(08)61637-4. [DOI] [PubMed] [Google Scholar]
  12. Nelson W. G., Kastan M. B. DNA strand breaks: the DNA template alterations that trigger p53-dependent DNA damage response pathways. Mol Cell Biol. 1994 Mar;14(3):1815–1823. doi: 10.1128/mcb.14.3.1815. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nunoshiba T., Demple B. Potent intracellular oxidative stress exerted by the carcinogen 4-nitroquinoline-N-oxide. Cancer Res. 1993 Jul 15;53(14):3250–3252. [PubMed] [Google Scholar]
  14. Ochman H., Gerber A. S., Hartl D. L. Genetic applications of an inverse polymerase chain reaction. Genetics. 1988 Nov;120(3):621–623. doi: 10.1093/genetics/120.3.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sussman M., Sussman R. R. The regulatory program for UDPgalactose polysaccharide transferase activity during slime mold cytodifferentiation: requirement for specific synthesis of ribonucleic acid. Biochim Biophys Acta. 1965 Nov 8;108(3):463–473. doi: 10.1016/0005-2787(65)90038-9. [DOI] [PubMed] [Google Scholar]
  16. Szymkowski D. E., Deering R. A. Identification and characterization of a Dictyostelium discoideum ribosomal protein gene. Nucleic Acids Res. 1990 Aug 25;18(16):4695–4701. doi: 10.1093/nar/18.16.4695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wilson D. M., 3rd, Deutsch W. A., Kelley M. R. Cloning of the Drosophila ribosomal protein S3: another multifunctional ribosomal protein with AP endonuclease DNA repair activity. Nucleic Acids Res. 1993 May 25;21(10):2516–2516. doi: 10.1093/nar/21.10.2516. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Xue Z., Mélèse T. Nucleolar proteins that bind NLSs: a role in nuclear import or ribosome biogenesis? Trends Cell Biol. 1994 Dec;4(12):414–417. doi: 10.1016/0962-8924(94)90095-7. [DOI] [PubMed] [Google Scholar]
  19. Zhou Z., Elledge S. J. DUN1 encodes a protein kinase that controls the DNA damage response in yeast. Cell. 1993 Dec 17;75(6):1119–1127. doi: 10.1016/0092-8674(93)90321-g. [DOI] [PubMed] [Google Scholar]

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