Reconstitution analysis of mating type switching recombination of fission yeast Schizosaccharomyces pombe

裂殖酵母交配型转换重组的重建分析

• 批准号: 21770007
• 负责人: MURAYAMA Yasuto
• 金额: $ 2.91万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21770007/
• 关键词: 接合型変換; 相同組換え; 遺伝子変換; Swi2-Swi5複合体; 分裂酵母; 試験管内; 再構成; D-loop形成; 生化学; 試験管内再構成; 接合; ヘテロクロマチン; Rad51; mat座位; 非対称分裂

In fission yeast Schizosaccharomyces pombe, mating type switching occurs at mat locus by using a mechanism of homologous recombination. The Swi2-Swi5 protein complex is considered to play an important role in the initiation of gene conversion during mating type switching. To reveal the biochemical function of the complex, I purified the Swi2-Swi5 protein complex, analyzed its biochemical activities and brought two important findings from the study. The first, Swi2 has at least two DNA binding domains; one is located at the N-terminal region which contains AT hook motif, and the other is located at the C-terminal region. The second, the Swi2-Swi5 complex promotes D-loop formation, which is an essential activity for initiation of strand exchange reaction. An N-terminal truncated Swi2 protein complexed with Swi5, which lacks the N-terminal DNA binding domain, promotes still D-loop formation, indicating that The C-terminal region of Swi2 has the essential domain for the D-loop activity. These results strongly suggest that Swi2-Swi5 directly involved in strand exchange reaction during mating type switching.

在裂殖酵母裂殖酵母中，交配型转换通过使用同源重组机制在mat位点发生。Swi 2-Swi 5蛋白复合物被认为在交配型转换期间基因转换的启动中起重要作用。为了揭示该复合物的生化功能，我纯化了Swi 2-Swi 5蛋白复合物，分析了其生化活性，并从研究中获得了两个重要发现。Swi 2至少有两个DNA结合结构域，一个位于N-末端，含有AT钩基序，另一个位于C-末端。第二，Swi 2-Swi 5复合物促进D环形成，这是引发链交换反应的必要活性。一个N-末端截短的Swi 2蛋白与Swi 5复合，缺乏N-末端DNA结合结构域，促进D-环的形成，表明Swi 2的C-末端区域具有D-环活性的必需结构域。这些结果强烈表明Swi 2-Swi 5直接参与交配型转换过程中的链交换反应。

项目成果

The fission yeast meiosis-specific Dmc1 recombinase mediates formation and branch migration of Holliday junction by preferentially promoting strand exchange in a direction opposite to that of Rad51

裂殖酵母减数分裂特异性 Dmc1 重组酶通过优先促进与 Rad51 方向相反的链交换来介导霍利迪连接体的形成和分支迁移

• 期刊: GENES and DEVELOPMENT誌 in press
• 作者: Yasuto Murayama;Yasuhiro Tsutsui;Hiroshi Iwasaki.
• 通讯作者: Hiroshi Iwasaki.

Regulation of Dmc1 recombinase by recombination mediator proteins.

重组介导蛋白对 Dmc1 重组酶的调节。

• 发表时间: 2009
• 作者: Yasuto Murayama;Yumiko Kurokawa;Hiroshi Iwasaki
• 通讯作者: Hiroshi Iwasaki

相同組換えメディエーターによるDmc1リコンビナーゼの正と負の制御

同源重组介质对 Dmc1 重组酶的正向和负向调节

• 发表时间: 2009
• 作者: 村山泰斗;黒川裕美子;岩崎博史
• 通讯作者: 岩崎博史

Biochemical characterization of regulation of Dmc1 recombinase by recombination mediator proteins.

重组介导蛋白对 Dmc1 重组酶调节的生化特征。

• 发表时间: 2009
• 作者: Yasuto Murayama;Yumiko Kurokawa;Hiroshi Iwasaki
• 通讯作者: Hiroshi Iwasaki

Biochemical characterization of regulation of Dmc1 recombinase by recombination mediator proteins

重组介体蛋白调节 Dmc1 重组酶的生化特征

• 发表时间: 2009
• 作者: 村山泰斗(発表者);黒川裕美子;岩崎博史
• 通讯作者: 岩崎博史