Biochemical characterization of apurinic/apyrimidinic endonuclease Triticum aestivum
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Keywords:
AP-endonuclease, TaAPE1L, wheat.Abstract
The gene encoding putative AP-endonuclease of Triticum aestivum was identified by homology to human and Arabidopsis AP-endonucleases by application of tBLASTn NCBI database search tool. For the first time cDNA of the wheat AP-endonuclease gene was isolated by reverse transcription-PCR. Functional expression of TaAPE1L in E.coli was performed and the protein was purified with nickel-based affinity chromatography to homogenous state. Through use of 5’-[P32] – marked oligonucleotide duplexes containing single tetrahydrofuran (analogue of AP-site) AP-endonuclease activity of TaAPE1L was shown. TaApe1L-catalyzed AP endonuclease activity increases depending on incubation time and that addition of EDTA in the reaction mixture completely blocks AP endonuclease activity of TaApe1LReferences
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2 Saparbaev M., Laval J. 3,N4-ethenocytosine, a highly mutagenic adduct, is a primary substrate for E. coli double-stranded uracil-DNA glycosylase and human mismatch-specific thymine-DNA glycosylase // Proc. Natl. Acad. Sci. – 1998. – Vol. 95. – P. 8508-8513.
3 Ischenko A.A., Saparbaev M.K. Alternative nucleotide incision repair pathway for oxidative DNA damage // Nature. – 2002. –Vol. 415. – Р. 183-187.
4 Cordoba-Canero D., Roldan-Arjona T., Ariza R. Arabidopsis ARP endonuclease functions in a branched base excision DNA repair pathway completed by LIG1 // Plant J. – 2011. – №68. – P.693-702
5 Zharkov D.O. Base excision DNA repair // Cell Mol .Life Sci. – 2008. – №65. -P.1544-1565
6 Sancar A., Lindsey-Boltz L., Unsal-Kacmaz K., Linn S. Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints // Annu. Rev. Biochem. – 2004. – №73. – P.39-85.
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Bissenbaev, A. K., Zholdybaeva, B. S., Tusumkhanova, Z. T., Akishev, Z. D., & Saparbaev, M. K. (2015). Biochemical characterization of apurinic/apyrimidinic endonuclease Triticum aestivum. Experimental Biology, 60(1), 27–30. Retrieved from https://bb.kaznu.kz/index.php/biology/article/view/49
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Bioinformatics, Genomics and Proteomics. Physical-chemical Biology
