Homing directed by VDE endonuclease in Saccharomyces cerevisiae

酿酒酵母中 VDE 核酸内切酶指导的归巢

• 批准号: 10216203
• 负责人: OHYA Yoshikazu
• 金额: $ 23.94万
• 依托单位: The University of Tokyo
• 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: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10216203/
• 关键词: VDE; Saccharomyces cerecvisiae; Non-homologous end joining; DNA repair; 遺伝的組み換え; ホーミング; 遺伝子ホーミング; VDE; 減数分裂; DNA切断; 核移行; 細胞死効果; 利己的遺伝子; ホーミングエンドヌクレアーゼ

In Saccharomyces cerevisiae, DNA double strand breaks (DSBs) are repaired by several mechanisms. Non-homologous end joining (NHEJ) efficiency is down-regulated in diploid cells relative to haploid cells at mitosis, but the mechanism of this mating type regulation at meiosis is unknown. When DSBs occur without homologous, template at meiosis, a few spores survive. In survival spores, DSBs might be repaired by NHEJ. To test this idea, we introduced DSB specifically in mieiosis by VDE endonuclease. and examined spore viability in the wild-type, nejl Δ and yku70Δ strains. The spore viability of these deletion mutants reduced in comparison with wild type. The viable spores contain short insertions or deletions typical for DNA repair caused by NHEL. The results suggest that DSBs is repaired by NHEJ in survival spores and DSBs repair by NHEJ occurs at meiosis.

在酿酒酵母中，DNA双链断裂（DSB）通过几种机制修复。非同源末端连接（NHEJ）效率在二倍体细胞中相对于在有丝分裂时的单倍体细胞下调，但在减数分裂时这种交配型调节的机制尚不清楚。当DSB在减数分裂时没有同源模板时，少数孢子存活。在存活的孢子中，DSB可能被NHEJ修复。为了验证这一想法，我们通过VDE内切酶将DSB特异性地引入到减数分裂中。并检查野生型、nejl Δ和yku70Δ菌株中的孢子活力。与野生型相比，这些缺失突变体的孢子活力降低。活孢子含有NHEL引起的DNA修复的典型短插入或缺失。结果表明，在存活孢子中，NHEJ修复DSB，NHEJ修复DSB发生在减数分裂时。

项目成果

Nagai, Y: "Karyopherin-mediated Nuclear Import of the Homing Endonuclease, VDE, Is Required for Self-propagation of the Coding Region."Mol.Cell Biol.. 23. 1726-1736 (2003)

Nagai, Y：“编码区域的自我繁殖需要核转运蛋白介导的归巢核酸内切酶 (VDE) 的核导入。”Mol.Cell Biol.. 23. 1726-1736 (2003)

Nogami, S: "Homing at an Extragenic locus Mediated by VDE (PI-Scel) of Saccharomyces cerevisiae"Yeast. 19. 773-782 (2002)

Nogami, S：“酿酒酵母 VDE (PI-Scel) 介导的外源位点归巢”酵母。

Okuda, Y: "Occurrence, horizonatal transfer and degeneration of VDE intein family in Saccharomycete yeasts"Yeast. (in press). (2003)

Okuda, Y：“酵母中 VDE 内含肽家族的发生、水平转移和退化”酵母。

Homma,K.: "Phosphatidylinositol-4-Phosphate 5-Kinase Localized on the plasma Membrane Is Essential for Yeast Cell Morphogenesis." J.Biol.Chem.273. 15779-15786 (1998)

Homma,K.：“位于质膜上的磷脂酰肌醇 4 磷酸 5 激酶对于酵母细胞形态发生至关重要。”

Mizutani, R: "Protein-splicing reaction via a thiazolidine intermediate : crystal structure of the VMA1-derived endonuclease bearing the N-and C-terminal propeptides"J.Mol.Biol.. 316. 919-929 (2002)

Mizutani, R：“通过噻唑烷中间体进行的蛋白质剪接反应：带有 N 端和 C 端前肽的 VMA1 衍生核酸内切酶的晶体结构”J.Mol.Biol.. 316. 919-929 (2002)