Screening of bio-active substances by using a peculiar phenotype of yeast mutant

利用酵母突变体特有的表型筛选生物活性物质

• 批准号: 11556017
• 负责人: MIYAKAWA Tokichi
• 金额: $ 7.87万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11556017/
• 关键词: Ca2+ signal; Calcineurin; Drug screening; MAP kinase; 生理活性物質; 出芽酵母; カルシニューリンニューリン; MAPキナーゼ; 細胞周期; カルシウム; 免疫抑制剤

An inappropriate activation of a signaling pathway in yeast often has a deleterious physiological effect and causes various defects, including growth defects. In a certain genetic background (Δzdsl) of Saccharomyces cerevisiae, the cell-cycle progression in G2 is specifically blocked in the medium with CaCl_2 by the hyperactivation of the Ca^<2+>-signaling pathways. Here, we developed a novel drug screening procedure designed to detect the active compounds that specifically attenuate the Ca^<2+>-signaling activity on the basis of the ability to abrogate the growth defect of the cells suffering from the hyperactivated Ca^<2+> signal. Using known calcineurin inhibitors as model compounds, we have established the screening conditions for the drugs that suppress the Ca^<2+>-induced growth inhibition. An indicator strain with an increased drug sensitivity was constructed with a syr1/erg3 null mutation.

酵母中信号通路的不适当激活通常具有有害的生理效应并导致各种缺陷，包括生长缺陷。在一定遗传背景（Δ zeta）的酿酒酵母（Saccharomycescerevisiae）中，G2期的细胞周期进程在CaCl_2培养基中被特异性阻断，这是由于Ca^<2+>-信号通路的过度激活所致。在此，我们开发了一种新的药物筛选程序，旨在检测特异性减弱Ca^2+信号活性的活性化合物，该活性化合物基于消除遭受过度活化的Ca^2+信号的细胞的生长缺陷的能力。以已知的钙调磷酸酶抑制剂为模型化合物，我们建立了抑制Ca^2+诱导的生长抑制的药物筛选条件。构建了具有syr 1/erg 3无效突变的药物敏感性增加的指示菌株。

项目成果

Masaki Mizunuma et al.: "GSK-3 kinase Mck1 and calcineurin coordinately mediates Hsl1 down-regulation by Ca^<2+> in budding yeast"EMBO J.. 20. 1074-1085 (2001)

Masaki Mizunuma 等人：“GSK-3 激酶 Mck1 和钙调神经磷酸酶在芽殖酵母中协调介导 Ca^2 下调 Hsl1”EMBO J.. 20. 1074-1085 (2001)

K. Kakugawa et al.: "Isolation of yeast Kurtzumanomyces sp. I-11, novel producer of mannosylerythritol lipid"Biosci. Blotechnol. Biochem.. 66. 188-191 (2002)

K. Kakukawa 等人：“酵母 Kurtzumanomyces sp. I-11 的分离，甘露糖赤藓糖醇脂质的新型生产者”Biosci。

Rota Tsujii et al.: "Ebp2, yeast homologue of a human protein that interacts with Epstein-Barr virus nuclear antigen 1, is required for pre-rRNA processing and ribosomal subunit assembly"Genes to Cells. 5. 543-553 (2000)

Rota Tsujii 等人：“Ebp2 是一种与 Epstein-Barr 病毒核抗原 1 相互作用的人类蛋白质的酵母同源物，是前 rRNA 加工和核糖体亚基组装所必需的”Genes to Cells。

Zhifeng Cui et al.: "Functional analysis of the promoter of the yeast SNQ2 gene encording a multidrug resistance transporter thai confers the resistance to 4-Nitroquinoline N-Oxide,"Biosci, Biotechnol. Biochem.. 63. 162-167 (1999)

崔志峰等人：“编码多药抗性转运蛋白的酵母 SNQ2 基因启动子的功能分析，该转运蛋白赋予对 4-硝基喹啉 N-氧化物的抗性”，Biosci，Biotechnol。

K. Kakugawa et al.: "Cloning, characterization, and expression of cDNA encoding a lipase from Kurtzumanomyces sp. I-11"Biosci. Biotechnot. Biochem.. 66. 1328-1336 (2002)

K. Kakukawa 等人：“编码来自 Kurtzumanomyces sp. I-11 的脂肪酶的 cDNA 的克隆、表征和表达”Biosci。