A Role for Saccharomyces cerevisiae Tpa1 Protein in Direct Alkylation Repair*
A Role for Saccharomyces cerevisiae Tpa1 Protein in Direct Alkylation Repair*
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酿酒酵母 Tpa1 蛋白在直接烷基化修复中的作用*
DOI：
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发表时间：
2014
影响因子：
4.8
通讯作者：
Roy Anindya
中科院分区：
文献类型：
--
作者：
Gururaj Shivange;Naveena Kodipelli;M. Monisha;Roy Anindya
背景：Fe(II)/2-酮戊二酸 (2OG) 依赖性双加氧酶修复 DNA 的烷基碱基损伤。结果：Tpa1是一种Fe(II)/2OG依赖性双加氧酶，在酿酒酵母中介导烷基碱基修复，用DNA糖基化酶MAG1删除TPA1对甲基化诱导的毒性敏感性具有协同作用。结论：Tpa1蛋白在DNA烷基化修复中发挥着至关重要的作用。意义：我们的结果提供了酿酒酵母中任何 Fe(II)/2OG 依赖性双加氧酶直接烷基化修复的第一个证据。烷化剂诱导细胞毒性 DNA 碱基加合物。在这项工作中，我们首次提供证据表明酿酒酵母 Tpa1 蛋白参与 DNA 烷基化修复。除了高通量分析的初步观察结果表明，TPA1 的缺失会导致酿酒酵母对甲磺酸甲酯敏感之外，人们对 Tpa1 作为修复蛋白知之甚少。使用纯化的 Tpa1，我们证明 Tpa1 可以修复单链和双链甲基化 DNA。 Tpa1 是 Fe(II) 和 2-酮戊二酸依赖性双加氧酶家族的成员，我们发现参与辅因子结合的氨基酸残基的突变会消除 Tpa1 DNA 修复活性。 TPA1 以及碱基切除修复途径 DNA 糖基化酶 MAG1 的缺失使得 tpa1Δmag1Δ 双突变体对甲基化诱导的毒性高度敏感。我们进一步证明，跨损伤合成 DNA 聚合酶 Pol z (REV3) 在耐受 DNA 甲基碱基损伤中发挥着关键作用，并且 tpa1Δmag1revΔ3 三重突变体对甲基化诱导的毒性极其敏感。我们的结果表明酿酒酵母中碱基切除修复途径和 Tpa1 直接烷基化修复之间存在协同作用。我们得出的结论是，Tpa1 是酵母中迄今为止尚未鉴定的 DNA 修复蛋白，它在恢复烷基化 DNA 碱基损伤和细胞毒性方面发挥着至关重要的作用。
Background: Fe(II)/2-oxoglutarate (2OG)-dependent dioxygenases repair alkyl-base lesions of DNA. Results: Tpa1 is a Fe(II)/2OG-dependent dioxygenase and mediates alkyl-base repair in Saccharomyces cerevisiae, and deleting TPA1 with DNA glycosylase MAG1 had a synergistic effect on the susceptibility to methylation-induced toxicity. Conclusion: Tpa1 protein plays a crucial role in DNA alkylation repair. Significance: Our results provide the first evidence of direct alkylation repair by any Fe(II)/2OG-dependent dioxygenases in Saccharomyces cerevisiae. 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 in DNA alkylation 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 double-stranded 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 tpa1Δmag1Δ double mutant highly susceptible to methylation-induced toxicity. We further demonstrate that the trans-lesion synthesis DNA polymerase Polζ (REV3) plays a key role in tolerating DNA methyl-base lesions and that tpa1Δmag1revΔ3 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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影响因子：
16
作者：
Ougland, R;Zhang, CM;Falnes, PO
通讯作者：
Falnes, PO
DOI：
10.1073/pnas.0403489101
发表时间：
2004-09-28
影响因子：
11.1
作者：
Delaney, JC;Essigmann, JM
通讯作者：
Essigmann, JM
能够修复烷基化 DNA 的布氏锥虫 Alkb 同源物的表征。
影响因子：
2.1
作者：
Simmons,JanaM;Koslowsky,DonnaJ;Hausinger,RobertP
通讯作者：
Hausinger,RobertP
影响因子：
50.3
作者：
Isaacs, JS;Jung, YJ;Neckers, L
通讯作者：
Neckers, L
酿酒酵母 DNA 聚合酶 eta 绕过胸腺嘧啶-胸腺嘧啶二聚体的准确性。
DOI：
10.1073/pnas.97.7.3094
发表时间：
2000
影响因子：
11.1
作者：
Washington,MT;Johnson,RE;Prakash,S;Prakash,L
通讯作者：
Prakash,L