%0 Journal Article
%T Cloning and Characterization of a Wheat Homologue of Apurinic/Apyrimidinic Endonuclease Ape1L
%A Botagoz Joldybayeva
%A Paulina Prorok
%A Inga R. Grin
%A Dmitry O. Zharkov
%A Alexander A. Ishenko
%A Barbara Tudek
%A Amangeldy K. Bissenbaev
%A Murat Saparbaev
%J PLOS ONE
%D 2014
%I Public Library of Science (PLoS)
%R 10.1371/journal.pone.0092963
%X Background Apurinic/apyrimidinic (AP) endonucleases are key DNA repair enzymes involved in the base excision repair (BER) pathway. In BER, an AP endonuclease cleaves DNA at AP sites and 3¡ä-blocking moieties generated by DNA glycosylases and/or oxidative damage. A Triticum aestivum cDNA encoding for a putative homologue of ExoIII family AP endonucleases which includes E. coli Xth, human APE1 and Arabidopsis thaliana AtApe1L has been isolated and its protein product purified and characterized. Methodology/Principal Findings We report that the putative wheat AP endonuclease, referred here as TaApe1L, contains AP endonuclease, 3¡ä-repair phosphodiesterase, 3¡ä-phosphatase and 3¡ä¡ú5¡ä exonuclease activities. Surprisingly, in contrast to bacterial and human AP endonucleases, addition of Mg2+ and Ca2+ (5¨C10 mM) to the reaction mixture inhibited TaApe1L whereas the presence of Mn2+, Co2+ and Fe2+ cations (0.1¨C1.0 mM) strongly stimulated all its DNA repair activities. Optimization of the reaction conditions revealed that the wheat enzyme requires low divalent cation concentration (0.1 mM), mildly acidic pH (6¨C7), low ionic strength (20 mM KCl) and has a temperature optimum at around 20¡ãC. The steady-state kinetic parameters of enzymatic reactions indicate that TaApe1L removes 3¡ä-blocking sugar-phosphate and 3¡ä-phosphate groups with good efficiency (kcat/KM = 630 and 485 ¦ÌM£¿1¡¤min£¿1, respectively) but possesses a very weak AP endonuclease activity as compared to the human homologue, APE1. Conclusions/Significance Taken together, these data establish the DNA substrate specificity of the wheat AP endonuclease and suggest its possible role in the repair of DNA damage generated by endogenous and environmental factors.
%U http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0092963