Characterization of the novel human AP endonuclease and poly-ADP-ribosyl transferase.

新型人 AP 核酸内切酶和聚 ADP 核糖基转移酶的表征。

• 批准号: 23510059
• 负责人: KANNO Shin-ichiro
• 金额: $ 3.49万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2013
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23510059/
• 关键词: DNA修復; APエンドヌクレアーゼ; PARP1; ポリADPリボシル化; アセチルトランスフェラーゼ

The CYR motif was first identified in the repair protein, PALF (ALPF), and found in various proteins of DNA metabolism and DNA repair. We newly identified the CYR motif containing proteins, Drosophila CG1218 and its human ortholog C4orf27, which contained a conserved DUF2228 domain. We hypothesized that these proteins have DNA metabolism related functions. We isolated cDNA of CG1218 and human C4orf27, and characterized these proteins. Here we show that; 1) a CG1218 protein accumulates at DNA damaged site and human C4orf27 protein forms a hetero dimmer with PARP1 and accumulates at DNA damaged site 2) CG1218 and C4orf27 possess endo/exonuclease activities against abasic site. 3) Suppression of the expression of C4orf27 using siRNA provided the cells with sensitivity to MMS, which produces methylated bases leading to single-strand breaks. These data suggest that DUF2228 protein CG1218-PA and C4orf27 play important roles in the repair of DNA base damage and single-strand breaks.

CYR基序首先在修复蛋白PALF（ALPF）中被发现，并在各种DNA代谢和DNA修复蛋白中发现。我们新鉴定了含有CYR基序的蛋白质，果蝇CG 1218及其人类直系同源物C4 orf 27，其中含有保守的DUF 2228结构域。我们推测这些蛋白质具有与DNA代谢相关的功能。我们分离了CG 1218和人C4 orf 27的cDNA，并对这些蛋白进行了表征。在这里，我们表明，1）CG 1218蛋白在DNA损伤位点处积累，并且人C4 orf 27蛋白与PARP 1形成异源二聚体并在DNA损伤位点处积累。2）CG 1218和C4 orf 27具有针对脱碱基位点的内切/外切核酸酶活性。3)使用siRNA抑制C4 orf 27的表达为细胞提供了对MMS的敏感性，MMS产生甲基化碱基，导致单链断裂。这些数据表明DUF 2228蛋白CG 1218-PA和C4 orf 27在DNA碱基损伤和单链断裂的修复中起重要作用。

项目成果

PALF acts as both a single-stranded DNA endonuclease and a single-stranded DNA 3’-exonuclease and can participate in DNA end joining in a biochemical system.

PALF既充当单链DNA核酸内切酶又充当单链DNA 3-核酸外切酶，并且可以参与生化系统中的DNA末端连接。

• 发表时间: 2011
• 期刊: J Biol Chem
• 影响因子: 4.8
• 作者: Ogiwara H;Ui A;Shiotani B;Zou L;Yasui A;Kohno T;Ogiwara H;Oike T;Ogiwara H et al;Ogiwara H;Li S et al
• 通讯作者: Li S et al

Mutations in UVSSA cause UV-sensitive syndrome and destabilize ERCC6 in transcription-coupled DNA repair

• DOI: 10.1038/ng.2228
• 发表时间: 2012-05-01
• 期刊: NATURE GENETICS
• 影响因子: 30.8
• 作者: Zhang, Xue;Horibata, Katsuyoshi;Tanaka, Kiyoji
• 通讯作者: Tanaka, Kiyoji

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