Direct observation of DNA rotation during branch migration of Holliday junction DNA by Escherichia coli RuvA-RuvB protein complex

Han YW; Tani T; Hayashi M; Hishida T; Iwasaki H; Shinagawa H; Harada Y

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Direct observation of DNA rotation during branch migration of Holliday junction DNA by Escherichia coli RuvA-RuvB protein complex

机译：通过大肠杆菌RuvA-RuvB蛋白复合物直接观察霍利迪结DNA分支迁移过程中DNA旋转

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The Escherichia coli RuvA-RuvB complex promotes branch migration of Holliday junction DNA, which is the central intermediate of homologous recombination. Like many DNA motor proteins, it is suggested that RuvA-RuvB promotes branch migration by driving helical rotation of the DNA. To clarify the RuvA-RuvB-mediated branch migration mechanism in more detail, we observed DNA rotation during Holliday junction branch migration by attaching a bead to one end of cruciform DNA that was fixed to a glass surface at the opposite end. Bead rotation was observed when RuvA, RuvB, and ATP were added to the solution. We measured the rotational rates of the beads caused by RuvA-RuvB-mediated branch migration at various ATP concentrations. The data provided a K-m value of 65 mu M and a V-max value of 1.6 revolutions per second, which corresponds to 8.3 bp per second. This real-time observation of the DNA rotation not only allows us to measure the kinetics of the RuvA-RuvB-mediated branch migration, but also opens the possibility of elucidating the branch migration mechanism in detail.

机译：大肠杆菌RuvA-RuvB复合物促进霍利迪连接DNA的分支迁移，霍利迪连接DNA是同源重组的核心中间产物。像许多DNA马达蛋白一样，有人建议RuvA-RuvB通过驱动DNA的螺旋旋转来促进分支迁移。为了更详细地阐明RuvA-RuvB介导的分支迁移机制，我们在霍利迪结分支迁移过程中观察了DNA旋转，方法是将珠子连接到十字形DNA的一端，该一端固定在另一端的玻璃表面上。当将RuvA，RuvB和ATP添加到溶液中时，观察到珠子旋转。我们测量了在各种ATP浓度下由RuvA-RuvB介导的分支迁移引起的珠的旋转速率。数据提供的K-m值为65μM，V-max值为每秒1.6转，相当于每秒8.3 bp。对DNA旋转的这种实时观察不仅使我们能够测量RuvA-RuvB介导的分支迁移的动力学，而且还提供了详细阐明分支迁移机制的可能性。

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《Proceedings of the National Academy of Sciences of the United States of America》 |2006年第31期|p. 11544-11548|共5页
作者
Han YW; Tani T; Hayashi M; Hishida T; Iwasaki H; Shinagawa H; Harada Y;
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作者单位

Tokyo Metropolitan Inst Med Sci, Bunkyo Ku, Tokyo 1138613, Japan;

Osaka Univ, Res Inst Microbial Dis, Suita, Osaka 5650871, Japan;

Yokohama City Univ, Grad Sch Integrated Sci, Tsurumi Ku, Yokohama, Kanagawa 2300045, Japan;

JST, CREST, Kawaguchi, Saitama 3320012, Japan;

BioAcad Inc, Ibaraki, Osaka 5670085, Japan;

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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种 eng
中图分类自然科学总论;
关键词
AAA(+) ATPase family; motor protein; single-molecule analysis; MOTOR PROTEIN; HOMOLOGOUS RECOMBINATION; RECOGNITION PROTEIN; CRYSTAL-STRUCTURE; SINGLE-MOLECULE; TRANSLOCATION; HELICASE; SEQUENCE; DOMAIN;

机译：AAA（+）ATPase家族;运动蛋白;单分子分析;运动蛋白;同源重组;识别蛋白;晶体结构;单分子;易位;螺旋酶;序列;域;

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专利

1. Recognition and manipulation of branched DNA by the RusA Holliday junction resolvase of Escherichia coli. [J] . Chan SN, Vincent SD, Lloyd RG Nucleic Acids Research . 1998,第7期

机译：大肠杆菌的RusA Holliday连接解离酶对分支DNA的识别和操纵。
2. Recognition and manipulation of branched DNA by the RusA Holliday junction resolvase of Escherichia coli [J] . Robert G. Lloyd, Sau N. Chan, Simon D. Vincent Nucleic acids research . 1998,第7期

机译：大肠杆菌的RusA Holliday连接解离酶对分支DNA的识别和操纵
3. Structural similarities between Escherichia coli RuvA protein and other DNA-binding proteins and a mutational analysis of its binding to the Holliday junction [J] . Rafferty JB., Hargreaves D., Artymiuk PJ., Journal of Molecular Biology . 1998,第1期

机译：大肠杆菌RuvA蛋白与其他DNA结合蛋白之间的结构相似性及其与霍利迪连接的结合的突变分析
4. Fluorimetric characterization of tryptophan residues in Escherichia coli single-stranded DNA-binding (SSB) protein and its poly(dT) complex [C] . Jose R. Casas-Finet Fluorescence Science and Technology . 2000

机译：大肠杆菌单链DNA结合（SSB）蛋白及其聚（DT）复合物中的色氨酸残基的荧光表征
5. Molecular studies in protein/DNA recognition: Crystallographic trials of the retrovirus integrase protein and the crystal structure of the Escherichia coli response regulator NarL(C)/DNA complex. [D] . Maris, Ann Elizabeth. 2002

机译：蛋白质/ DNA识别的分子研究：逆转录病毒整合酶蛋白的结晶学试验和大肠杆菌反应调节因子NarL（C）/ DNA复合物的晶体结构。
6. Direct observation of DNA rotation during branch migration of Holliday junction DNA by Escherichia coli RuvA–RuvB protein complex [O] . Yong-Woon Han, Tomomi Tani, Masahito Hayashi, 2006

机译：通过大肠杆菌RuvA–RuvB蛋白复合物直接观察霍利迪结DNA分支迁移过程中DNA旋转
7. Direct observation of DNA rotation during branch migration of Holliday junction DNA by Escherichia coli RuvA–RuvB protein complex [O] . Han, Yong-Woon, Tani, Tomomi, Hayashi, Masahito, 2006

机译：通过大肠杆菌RuvA–RuvB蛋白复合物直接观察霍利迪结DNA分支迁移过程中DNA旋转

Direct observation of DNA rotation during branch migration of Holliday junction DNA by Escherichia coli RuvA-RuvB protein complex

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