Molecular mechanisms of branch migration and resolution of Holliday junctions

霍利迪连接体分支迁移和分解的分子机制

• 批准号: 06404002
• 负责人: SHINAGAWA Hideo
• 金额: $ 15.74万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06404002/
• 关键词: DNA recombination; DNA repair; Holliday junction; Mutant; RecG protein; RuvA protein; RuvB protein; RuvC protein; RuvCエンドヌクレアーゼ; RuvGタンパク質; RuvCタンパク; エンドヌクレアーゼ; X線結晶回折; 染色体

We studied reaction mechanisms and functions of RuvA and RuvB protein complex and RecG protein which drives branch migration of Holliday junctions and RuvC protein which endonucleolytically resolves Holliday junctions.1.We discovered that RecG protein has a unique function to resolve the R-loops formed at the replication origin and negatively regulate the initiation of replication.2.We studied the substrate specificity of RuvC resolvase with various synthetic Holliday junctions and found that the existence of thymine residue at or near crossover point is important but the sequence homology at the junction is not important for the cleavage.3.We cloned and analyzed the ruvA,ruvB and ruvC genes of Pseudomonas aeruginosa and found that although the arrangement of the genes are different from that of the Escherichia coli genes, they possess the similar functions.4.We cloned, expressed the Thermus thermophilus ruvB gene and characterized the RuvB protein.5.We studied the morphology of ruv mutants and recG mutants and found that the abortive recombination in these mutants inhibit the chromosome nondisjunction.

我们研究了RUVA和RUVB蛋白复合物和RECG蛋白的反应机制和功能，这些蛋白可以驱动霍利迪连接和RUVC蛋白的分支迁移，该分支迁移，核酸内核酸溶解溶解Holliday结合。1。我们发现，RECG蛋白具有独特的功能，可以解析出在R-loolops中的独特功能。复制起源和负面调节复制的开始。2。我们研究了用各种合成的霍利迪连接处的RUVC分解的底物特异性3.我们克隆并分析了铜绿假单胞菌的Ruva，Ruvb和Ruvc基因，发现该基因的排列与大肠杆菌基因的排列不同，它们具有相似的功能。 ，表达了热嗜热RUVB基因并表征了RUVB蛋白。5。我们研究了Ruv突变体和RECG突变体的形态，发现这些突变体中的流产重组抑制了染色体非分离染色体。

项目成果

品川日出夫: "臨床分子生物学(分担執筆:DNA修復と突然変異・癌)" 日本臨床, 6 (1994)

品川秀夫：“临床分子生物学（贡献者：DNA 修复和突变/癌症）”Nippon Clinical，6 (1994)

Hishida, T., Iwasaki, H., Ishioka, K. Shinagawa, H.: "Molecular analysis of the Pseudomonas aeryginosa genes, ruvA, ruvB and ruvC, involved in processing of homologous recombination intermediates." Gene. 182. 63-70 (1996)

Hishida, T.、Iwasaki, H.、Ishioka, K. Shinakawa, H.：“铜绿假单胞菌基因 ruvA、ruvB 和 ruvC 的分子分析，参与同源重组中间体的加工。”

Shida,T.,Iwasaki,H.,Saito,A.,Kyogoku,Y.,Shinagawa,H.: "Analysis of substrate specificity of the RuvC Holliday junction resolvase with synthetic Holiday junctions." J.Biol.Chem.271. 26105-26109 (1996)

Shida,T.、Iwasaki,H.、Saito,A.、Kyogoku,Y.、Shinakawa,H.：“使用合成假日连接点分析 RuvC Holliday 连接点解析酶的底物特异性。”

Ariyoshi, M., Vassylyev, D.G., Iwasaki, H., Nakamura, H., Shinagawa, H.and Morikawa, K.: "Atomic structure of the RuvC resolvase : a Holliday junction-specific endonuclease from E.coli." Cell. 78. 1063-1072 (1994)

Ariyoshi, M.、Vassylyev, D.G.、Iwasaki, H.、Nakamura, H.、Shinakawa, H. 和 Morikawa, K.：“RuvC 解离酶的原子结构：来自大肠杆菌的霍利迪连接特异性核酸内切酶。”

Tahahagi,M.,Iwasaki,H.,Shinagawa,H.: "Structual requirements of substrate DNA for binding to and cleavage by RuvC、 a Holliday junction resolvase.21GC02:J.Biol.Chem.," 269. 15132-15139 (1994)

Tahahagi, M.、Iwasaki, H.、Shinakawa, H.：“底物 DNA 与霍利迪连接解析酶 RuvC 结合和切割的结构要求。21GC02：J.Biol.Chem.”，269. 15132-15139（ 1994）