Structure and function of DNA repair enzymes

DNA修复酶的结构和功能

• 批准号: 05270101
• 负责人: YASUI Akira
• 金额: $ 93.7万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05270101/
• 关键词: DNA repair; protein structure; DNA damages; DNA binding protein; 修復酵素; 分子生物学; 遺伝情報; 遺伝子; 結晶化

In this research project a number of novel DNA repair genes and proteins were isolated and structures of various repair proteins were determined as listed below ;1) Cyclobutane pyrimidine dimer (CPD) photolyases from various higher eukaryotes including aplacental mammals and a 6-4photoproduct (64PP) photolyase gene from Drosophila melanogaster were firstly isolated. Novel UV endonuclease genes for CPD and 64PP was isolated from eukaryotes and a procaryote.2) APEX (AP endonuclease) gene was isolated from mouse and its roles in DNA repair and development were identified.3) A human gene encoding a RecQ homolog was isolated. Disruption of a yeast homolog of the cloned human gene gave a MMS sensitive, sporulation-deficient mutant.4) A novel photosensitive human cell line like a Cockayne's syndrome (CS) was identified.5) p53 protein was found to be induced in cell lines of CS of XPA with less UV dose than in XPC or HeLa cells. It was concluded that transcription block by UV damage activates p53.6) An in vitro assay for NER was established using SV40 mini chromosome and used to isolate XPC and HHR23 genes. Furthermore, the association of both gene products was found to be essential for NER activity.7) Structures of repair proteins, RUVC,XPA,T4 CPD glycosylase, 3-methyl adenine glycosylase and CPD photolyase, were firstly determined.

本研究分离了一系列新的DNA修复基因和蛋白，并确定了各种修复蛋白的结构，具体如下：1）首次从包括无胎盘哺乳动物在内的多种高等真核生物中分离到环丁烷嘧啶二聚体（CPD）光解酶基因，并从黑腹果蝇（Drosophilamelanogaster）中分离到一个6- 4光产物（64 PP）光解酶基因。从真核生物和原核生物中分离到CPD和64 PP的紫外核酸内切酶基因; 2）从小鼠中分离到APEX（AP内切酶）基因，并鉴定了其在DNA修复和发育中的作用; 3）分离到一个编码RecQ同源物的人基因。4）鉴定了一种新的光敏人类细胞系，如Cockayne综合征（CS）。5）发现在XPA的CS细胞系中，p53蛋白在比XPC或HeLa细胞更少的UV剂量下被诱导。6）利用SV 40微型染色体建立了NER的体外检测方法，并用于分离XPC和HHR 23基因。7）首次确定了修复蛋白RUVC、XPA、T4 CPD糖基化酶、3-甲基腺嘌呤糖基化酶和CPD光裂合酶的结构。

项目成果

Todo,T.: "A new photoreactivating enzyme that specifically repair ultraviolet light-induced (6-4) photoproducts" Nature. 361. 371-374 (1993)

Todo,T.：“一种新的光活化酶，专门修复紫外线诱导的 (6-4) 光产物”，《自然》。

Ariyoshi,M.: "Atomic structure of the RuvC resolvase:a Holliday junction specific endonuclease from E.coli." Cell. 76. 1063-1072 (1994)

Ariyoshi,M.：“RuvC 分解酶的原子结构：来自大肠杆菌的霍利迪连接体特异性核酸内切酶。”

Yasui,A.,: "Avew class of DNA photolyases present in various oganisms" EMBO J.13. 6143-6151 (1994)

Yasui,A.，：“各种生物体中存在的 Avew 类 DNA 光裂合酶”EMBO J.13。

Kimura,K.: "Growth state-and cell cycle-dependent fluchnation" J.Biol.Chem.269. 1173-1176 (1994)

Kimura,K.：“生长状态和细胞周期依赖性波动”J.Biol.Chem.269。

Yamaizumi,M.: "U.v.-induced nuclear accumulation of p53 is evoked through DNA damage of actively transcribed genes independent of the cell cycle." Oncogene. 9. 2775-2784 (1994)

Yamaizumi,M.：“紫外线诱导的 p53 核积累是通过独立于细胞周期的活跃转录基因的 DNA 损伤引起的。”