Structural biology of human REV1, REV3 and REV7 complex

人类 REV1、REV3 和 REV7 复合物的结构生物学

• 批准号: 20770089
• 负责人: HASHIMOTO Hiroshi
• 金额: $ 2.75万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20770089/
• 关键词: X線結晶解析; DNAポリメラーゼ; DNA複製; DNA損傷; 損傷乗り越えDNA合成; X線結晶構造解析; REV7; REV3; REV1; タンパク質複合体

DNA polymerase ζ (Polζ) is an error-prone DNA polymerase involved in translesion DNA synthesis (TLS). Polζ consists of two subunits: the catalytic REV3, which belongs to B-family DNA polymerase, and the non-catalytic REV7. REV7 also interacts with REV1 polymerase, which is an error-prone Y-family DNA polymerase and also involved in TLS. Cells deficient in one of the three REV proteins and those deficient in all three proteins show similar phenotype, indicating the functional collaboration of the three REV proteins. REV7 interacts with both REV3 and REV1 polymerases, but the structure of REV7 or REV3, as well as the structural and functional basis of the REV1-REV7 and REV3-REV7 interactions remains unknown. Here we show the first crystal structure of human REV7 in complex with a fragment of human REV3 polymerase (residues 1847-1898) and reveal the mechanism underlying REV7-REV3 interaction. The structure indicates that the interaction between REV7 and REV3 creates a structural interface for REV1-binding. Furthermore, we show that the REV7-mediated interactions are responsible for DNA-damage tolerance. Our results highlight the function of REV7 as an adapter protein to recruit Polζ to a lesion site. REV7 is alternatively called MAD2B or MAD2L2 and also involved in various cellular functions such as signal transduction and cell-cycle regulation. Our results will provide a general structural basis for understanding the REV7-interaction.

DNA聚合酶β（Polymerase β，Polymerase β）是一种参与跨损伤DNA合成（translesion DNA synthesis，TLS）的易错DNA聚合酶。聚合酶由两个亚基组成：催化REV 3，属于B家族DNA聚合酶，和非催化REV 7。REV 7还与REV 1聚合酶相互作用，REV 1聚合酶是一种易出错的Y家族DNA聚合酶，也参与TLS。缺乏三种REV蛋白中的一种的细胞和缺乏所有三种蛋白的细胞表现出相似的表型，表明三种REV蛋白的功能协作。REV 7与REV 3和REV 1聚合酶两者相互作用，但REV 7或REV 3的结构以及REV 1-REV 7和REV 3-REV 7相互作用的结构和功能基础仍然未知。在这里，我们展示了与人REV 3聚合酶片段（残基1847-1898）复合的人REV 7的第一个晶体结构，并揭示了REV 7-REV 3相互作用的机制。结构表明REV 7和REV 3之间的相互作用为REV 1结合创建了结构界面。此外，我们表明，REV 7介导的相互作用是负责DNA损伤耐受性。我们的研究结果强调了REV 7作为衔接蛋白将Pol β募集到病变部位的功能。REV 7也被称为MAD 2B或MAD 2L 2，也参与各种细胞功能，如信号转导和细胞周期调节。我们的研究结果将为理解REV 7相互作用提供一般的结构基础。

项目成果

Crystal structure of DNAADP-ribosylating protein, pierisin-1

DNAADP-核糖基化蛋白，pierisin-1 的晶体结构

• 发表时间: 2009
• 作者: T.Shimizu;H.Hashimoto;K.Hiraga;T.Oda;T.Nakano;T.Sugimura;K.Wakabayashi;M.Sato
• 通讯作者: M.Sato

Crystal Structure of Human REV7 in Complex with a Human REV3 Fragment and Structural Implication of the Interaction between DNA Polymerase ζ and REV1

• DOI: 10.1074/jbc.m109.092403
• 发表时间: 2010-04-16
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Hara, Kodai;Hashimoto, Hiroshi;Sato, Mamoru
• 通讯作者: Sato, Mamoru

Structural basis for novel interactions between human TLS polymerases and PCNA

人类 TLS 聚合酶和 PCNA 之间新型相互作用的结构基础

• 发表时间: 2009
• 作者: Asami Hishiki;Hiroshi Hashimoto;Tomo Hanafusa;Keijiro Kamei;Eiji Ohashi;Toshiyuki Shimizu;Haruo Ohmori;Mamoru Sato
• 通讯作者: Mamoru Sato

Structural biology of a nuclear import of proteins by transportin 1

转运蛋白 1 向核输入蛋白质的结构生物学

• 发表时间: 2008
• 作者: M. Sato;T. Imasaki;T. Shimizu;H. Hashimoto;H. Ishida;D. Kamei M. Yamada
• 通讯作者: D. Kamei M. Yamada

The structure of the N-terminal regulatory domain of a plant NADPH oxidase and its functional implications

植物NADPH氧化酶N端调控域的结构及其功能意义

• 发表时间: 2010
• 期刊: J Biol Chem
• 影响因子: 4.8
• 作者: Oda T;Hashimoto H;Kuwabara N;Akashi S;Hayashi K;Kojima C;Wong HL;Kawasaki T;Shimamoto K;Sato M;Shimizu T
• 通讯作者: Shimizu T