A role for saccharomyces cerevisiae Tpa1 protein in direct alkylation repair

Shivange, G and Kodipelli, N and M, Monisha and Roy, Anindya (2014) A role for saccharomyces cerevisiae Tpa1 protein in direct alkylation repair. Journal of Biological Chemistry (289). pp. 35939-35952. ISSN 0021-9258

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Alkylating agents induce cytotoxic DNA base adducts. In this work, we provide evidence to suggest, for the first time, that Saccharomyces cerevisiae Tpa1 protein is involved inDNAalkylation repair. Little is known about Tpa1 as a repair protein beyond the initial observation from a high-throughput analysis indicating that deletion of TPA1 causes methyl methane sulfonate sensitivity in S. cerevisiae. Using purified Tpa1, we demonstrate that Tpa1 repairs both single- and doublestranded methylated DNA. Tpa1 is a member of the Fe(II) and 2-oxoglutarate-dependent dioxygenase family, and we show that mutation of the amino acid residues involved in cofactor binding abolishes the Tpa1 DNA repair activity. Deletion of TPA1 along with the base excision repair pathway DNA glycosylase MAG1 renders the tpa1▵mag1▵ double mutant highly susceptible to methylation-induced toxicity. We further demonstrate that the trans-lesion synthesis DNA polymerase Pol- (REV3) plays a key role in tolerating DNA methyl-base lesions and that tpa1▵mag1rev▵3 triple mutant is extremely susceptible to methylation-induced toxicity. Our results indicate a synergism between the base excision repair pathway and direct alkylation repair by Tpa1 in S. cerevisiae. We conclude that Tpa1 is a hitherto unidentified DNA repair protein in yeast and that it plays a crucial role in reverting alkylated DNA base lesions and cytotoxicity.

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IITH Creators:
IITH CreatorsORCiD
Roy, Anindyahttp://orcid.org/0000-0001-8561-907X
Item Type: Article
Uncontrolled Keywords: Alkylation; Amino acids; Cytotoxicity; DNA; Methane; Methylation; Proteins; Toxicity; Yeast 2-oxoglutarate-dependent dioxygenase; Alkylating agents; Amino acid residues; Base excision repairs; Cofactor binding; Direct alkylation; DNA repair proteins; High-throughput analysis
Subjects: Others > Biotechnology
Divisions: Department of Biotechnology
Depositing User: Team Library
Date Deposited: 12 Jan 2015 05:20
Last Modified: 05 Dec 2017 04:53
URI: http://raiith.iith.ac.in/id/eprint/1311
Publisher URL: https://doi.org/10.1074/jbc.M114.590216
OA policy: http://www.sherpa.ac.uk/romeo/issn/0021-9258/
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