Abstract
Repair of the ubiquitous mutagenic lesion 7,8-dihydro-8-oxoguanine (8-oxoG) is initiated in eukaryotes by DNA glycosylases/lyases, such as yeast Ogg1, that do not share significant sequence identity with their prokaryotic counterparts, typified by Escherichia coli MutM (Fpg) protein. The unexpected presence of a functional mutM orthologue in the model plant Arabidopsis thaliana has brought into question the existence of functional OGG1 orthologues in plants. We report here the cDNA cloning, expression and functional characterization of AtOGG1, an Arabidopsis thaliana gene widely expressed in different plant tissues which encodes a 40.3 kDa protein with significant sequence identity to yeast and human Ogg1 proteins. Purified AtOgg1 enzyme specifically cleaves duplex DNA containing an 8-OxoG:C mispair, and the repair reaction proceeds through an imine intermediate characteristic of all bifunctional DNA glycosylases/lyases. Consistent with its in vitro activity, expression of AtOGG1 suppresses the mutator phenotype of an E. coli strain deficient in 8-oxoG repair. Our results suggest that AtOgg1 is an structural and functional homologue of Ogg1 and establish the presence of two distinct 8-oxoG repair enzymes in Arabidopsis.
Similar content being viewed by others
References
Aburatani, H., Hippo, Y., Ishida, T., Takashima, R., Matsuba, C., Kodama, T., Takao, M., Yasui, A., Yamamoto, K., Asano, M., Fukasawa, K., Yoshinari, T., Inoue, H., Ohtsuka, E. and Nishimura, S. 1997. Cloning and characterization of mammalian 8-hydroxyguanine-specific DNA glycosylase/apurinic, apyrim-idinic lyase, a functional MutM homolog. Cancer Res. 57: 2151–2156.
Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W. and Lipman, D.J. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl. Acids Res. 25: 3389–3402.
Arai, K., Morishita, K., Shinmura, K., Kohno, T., Kim, S.R., Nohmi, T., Taniwaki, M., Ohwada, S. and Yokota, J. 1997. Cloning of a human homolog of the yeast OGG1 gene that is involved in the repair of oxidative DNA damage. Oncogene 14: 2857–2861.
Bhagwat, M. and Gerlt, J.A. 1996. 3′-and5′-strand cleavage reac-tions catalyzed by the Fpg protein from Escherichia coli occur via successive β-andδ-elimination mechanisms, respectively. Biochemistry 35: 659–665.
Bjoras, M., Luna, L., Johnsen, B., Hoff, E., Haug, T., Rognes, T. and Seeberg, E. 1997. Opposite base-dependent reactions of a human base excision repair enzyme on DNA containing 7,8-dihydro-8-oxoguanine and abasic sites. EMBO J. 16: 6314–6322.
Boiteux, S., Gajewski, E., Laval, J. and Dizdaroglu, M. 1992. Substrate specificity of the Escherichia coli Fpg protein (formamidopyrimidine-DNA glycosylase): excision of purine lesions in DNA produced by ionizing radiation or photosensi-tization. Biochemistry 31: 106–110.
Bruner, S.D., Norman, D.P. and Verdine, G.L. 2000. Structural basis for recognition and repair of the endogenous mutagen 8-oxoguanine in DNA. Nature 403: 859–866.
Castaing, B., Geiger, A., Seliger, H., Nehls, P., Laval, J., Zelwer, C. and Boiteux, S. 1993. Cleavage and binding of a DNA fragment containing a single 8-oxoguanine by wild type and mutant FPG proteins. Nucl. Acids Res. 21: 2899–2905.
Cheng, K.C., Cahill, D.S., Kasai, H., Nishimura, S. and Loeb, L.A. 1992. 8-Hydroxyguanine, an abundant form of oxidative DNA damage, causes G→T and A→C substitutions. J. Biol. Chem. 267: 166–172.
Cokol, M., Nair, R. and Rost, B. 2000. Finding nuclear localization signals. EMBO Rep. 1: 411–415.
Deutsch, W.A., Yacoub, A., Jaruga, P., Zastawny, T.H. and Diz-daroglu, M. 1997. Characterization and mechanism of action of Drosophila ribosomal protein S3 DNA glycosylase activity for the removal of oxidatively damaged DNA bases. J. Biol. Chem. 272: 32857–32860.
Dherin, C., Dizdaroglu, M., Doerflinger, H., Boiteux, S. and Radi-cella, J.P. 2000. Repair of oxidative DNA damage in Drosophila melanogaster: identification and characterization of dOgg1, a second DNA glycosylase activity for 8-hydroxyguanine and formamidopyrimidines. Nucl. Acids Res. 28: 4583–4592.
Dizdaroglu, M. 1999. Mechanisms of oxidative DNA damage; lesions and their measurement. In: M. Dizdaroglu and A.E. Karakaya (Eds.) Advances in DNA Damage and Repair: Oxy-gen Radical Effects, Cellular Protection, and Biological Con-sequences, Kluwer Academic Publishers, Dordrecht, Netherlands Plenum, New York, pp. 67–87.
Dodson, M.L., Michaels, M.L. and Lloyd, R.S. 1994. Unified cat-alytic mechanism for DNA glycosylases. J. Biol. Chem. 269: 32709–32712.
Eisen, J.A. and Hanawalt, P.C. 1999. A phylogenomic study of DNA repair genes, proteins, and processes. Mutat. Res. 435: 171–213.
Girard, P.M., Guibourt, N. and Boiteux, S. 1997. The Ogg1 protein of Saccharomyces cerevisia e: a 7,8-dihydro-8-oxoguanine DNA glycosylase/AP lyase whose lysine 241 is a critical residue for catalytic activity. Nucl. Acids Res. 25: 3204–3211.
Karahalil, B., Girard, P.M., Boiteux, S. and Dizdaroglu, M. 1998. Substrate specificity of the Ogg1 protein of Saccharomyces cerevisia e: excision of guanine lesions produced in DNA by ion-izing radiation-or hydrogen peroxide/metal ion-generated free radicals. Nucl. Acids Res. 26: 1228–1233.
Krokan, H.E., Standal, R. and Slupphaug, G. 1997. DNA glyco-sylases in the base excision repair of DNA. Biochem. J. 325: 1–16.
Le Page, F., Kwoh, E.E., Avrutskaya, A., Gentil, A., Leadon, S.A., Sarasin, A. and Cooper, P.K. 2000. Transcription-coupled repair of 8-oxoguanine: requirement for XPG, TFIIH, and CSB and implications for Cockayne syndrome. Cell 101: 159–171.
Lindahl, T. 1993. Instability and decay of the primary structure of DNA. Nature 362: 709–715.
Lu, R., Nash, H.M. and Verdine, G.L. 1997. A mammalian DNA repair enzyme that excises oxidatively damaged guanines maps to a locus frequently lost in lung cancer. Curr. Biol. 7: 397–407.
Maki, H. and Sekiguchi, M. 1992. MutT protein specifically hy-drolyses a potent mutagenic substrate for DNA synthesis. Nature 355: 273–275.
McCullough, A.K., Dodson, M.L. and Lloyd, R.S. 1999. Initiation of base excision repair: glycosylase mechanisms and structures. Annu. Rev. Biochem. 68: 255–285.
Michaels, M.L., Cruz, C., Grollman, A.P. and Miller, J.H. 1992. Evidence that MutY and MutM combine to prevent mutations by an oxidatively damaged form of guanine in DNA. Proc. Natl. Acad. Sci. USA 89: 7022–7025.
Michaels, M.L. and Miller, J.H. 1992. The GO system pro-tects organisms from the mutagenic effect of the spontaneous lesion 8-hydroxyguanine (7,8-dihydro-8-oxoguanine). J. Bact. 174: 6321–6325.
Moriya, M. 1993. Single-stranded shuttle phagemid for mutagene-sis studies in mammalian cells: 8-oxoguanine in DNA induces targeted G.C →T.A transversions in simian kidney cells. Proc. Natl. Acad. Sci. USA 90: 1122–1126.
Moriya, M., Ou, C., Bodepudi, V., Johnson, F., Takeshita, M. and Grollman, A.P. 1991. Site-specific mutagenesis using a. gapped duplex vector: a study of translesion synthesis past 8-oxodeoxyguanosine in E. col i. Mutat. Res. 254: 281–288.
Nash, H.M., Bruner, S.D., Scharer, O.D., Kawate, T., Addona, T.A., Spooner, E., Lane, W.S. and Verdine, G.L. 1996. Cloning of a yeast 8-oxoguanine DNA glycosylase reveals the existence of a base-excision DNA-repair protein superfamily. Curr. Biol. 6: 968–980.
Ohtsubo, T., Matsuda, O., Iba, K., Terashima, I., Sekiguchi, M. and Nakabeppu, Y. 1998. Molecular cloning of AtMMH, an Ara-bidopsis thaliana ortholog of the Escherichia coli mutM gene, and analysis of functional domains of its product. Mol. Gen. Genet. 259: 577–590.
Radicella, J.P., Dherin, C., Desmaze, C., Fox, M.S. and Boiteux, S. 1997. Cloning and characterization of hOGG1, a human ho-molog of the OGG1 gene of Saccharomyces cerevisia e.Proc. Natl. Acad. Sci. USA 94: 8010–8015.
Roldan-Arjona, T., Wei, Y.F., Carter, K.C., Klungland, A., Anselmino, C., Wang, R.P., Augustus, M. and Lindahl, T. 1997. Molecular cloning and functional expression of a human cDNA encoding the antimutator enzyme 8-hydroxyguanine-DNA gly-cosylase. Proc. Natl. Acad. Sci. USA 94: 8016–8020.
Rosenquist, T.A., Zharkov, D.O. and Grollman, A.P. 1997. Cloning and characterization of a mammalian 8-oxoguanine DNA glyco-sylase. Proc. Natl. Acad. Sci. USA 94: 7429–7434.
Sakumi, K., Furuichi, M., Tsuzuki, T., Kakuma, T., Kawabata, S., Maki, H. and Sekiguchi, M. 1993. Cloning and expression of cDNA for a human enzyme that hydrolyzes 8-oxo-dGTP, a mutagenic substrate for DNA synthesis. J. Biol. Chem. 268: 23524–23530.
Shibutani, S., Takeshita, M. and Grollman, A.P. 1991. Insertion of specific bases during DNA synthesis past the oxidation-damaged base 8-oxodG. Nature 349: 431–434.
Slupska, M.M., Luther, W.M., Chiang, J.H., Yang, H. and Miller, J.H. 1999. Functional expression of hMYH, a human homolog of the Escherichia coli MutY protein. J. Bact. 181: 6210–6213.
Snead, M.A., Alting-Mess, M.A. and Short, J.M. 1997. Clone exci-sion methods for the Lambda ZAP-based vectors. In: I.G. Cowell and C.A. Austin (Eds.) Methods in Molecular Biology, Vol. 69: cDNA Library Protocols, Humana Press, Totowa, NJ, pp. 53–60.
Soni, R. and Murray, A.H. 1994. Isolation of intact DNA and RNA from plant tissues. Anal. Biochem. 218: 474–476.
Studier, F.W., Rosenberg, A.H., Dunn, J.J. and Dubendorff, J.W. 1990. Use of T7 RNA polymerase to direct expression of cloned genes. Meth. Enzymol. 185: 60–89.
Tchou, J., Kasai, H., Shibutani, S., Chung, M.H., Laval, J., Grollman, A.P. and Nishimura, S. 1991. 8-oxoguanine (8-hydroxyguanine) DNA glycosylase and its substrate specificity. Proc. Natl. Acad. Sci. USA 88: 4690–4694.
Thomas, D., Scot, A.D., Barbey, R., Padula, M. and Boiteux, S. 1997. Inactivation of OGG1 increases the incidence of G.C →T.A transversions in Saccharomyces cerevisia e: evidence for en-dogenous oxidative damage to DNA in eukaryotic cells. Mol. Gen. Genet. 254: 171–178.
Thompson, J.D., Higgins, D.G. and Gibson, T.J. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucl. Acids Res. 22: 4673–4680.
van der Kemp, P.A., Thomas, D., Barbey, R., de Oliveira, R. and Boiteux, S. 1996. Cloning and expression in Escherichia coli of the OGG1 gene of Saccharomyces cerevisia e, which codes for a DNA glycosylase that excises 7,8-dihydro-8-oxoguanine and 2,6-diamino-4-hydroxy-5-N-methylformamidopyrimidine. Proc. Natl. Acad. Sci. USA 93: 5197–5202.
Weigel, D., Alvarez, J., Smyth, D.R., Yanofsky, M.F. and Meyerowitz, E.M. 1992. LEAFY controls floral meristem iden-tity in Arabidopsi s. Cell 69: 843–859.
Wood, M.L., Dizdaroglu, M., Gajewski, E. and Essigmann, J.M. 1990. Mechanistic studies of ionizing radiation and oxidative mutagenesis: genetic effects of a single 8-hydroxyguanine (7-hydro-8-oxoguanine) residue inserted at a unique site in a viral genome. Biochemistry 29: 7024–7032.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
García-Ortiz, MV., Ariza, R.R. & Roldán-Arjona, T. An OGG1 orthologue encoding a functional 8-oxoguanine DNA glycosylase/lyase in Arabidopsis thaliana . Plant Mol Biol 47, 795–804 (2001). https://doi.org/10.1023/A:1013644026132
Issue Date:
DOI: https://doi.org/10.1023/A:1013644026132