Structural and functional analysis and its application of the yeast novel N-asetylransferase that detoxifies a proline analogue

脯氨酸类似物解毒酵母新型N-乙酰基转移酶的结构和功能分析及其应用

• 批准号: 15380076
• 负责人: TAKAGI Hiroshi
• 金额: $ 9.54万
• 依托单位: Fukui Prefectural University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380076/
• 关键词: Yeast; N-Acetyltrasnferase; Azetidine-2-carboxylic acid; Anti-oxidant enzyme; Oxidative stress; Reactive oxygen species; Freezing stress; Ethanol stress; プロリンアナログ耐性

The MPR1 gene (sigma 1278b gene for proline-analogue resistance) of the budding yeast Saccharomyces cerevisiae, which is found in the Σ1278b strain, encodes a novel N-acetyltransferase (Mpr1) that detoxifies the proline analogue azetidine-2-carboxylate (AZC). In this study, we analyzed the structure and function of Mpr1 as follows.1)We found that mpr1-disrupted cells were hypersensitive to oxidative stresses such as H_2O_2 and heat shock and contained increased levels of reactive oxygen species (ROS). In contrast, expression of MPR1 leads to a decrease in ROS level and an increase in cell viability after oxidative treatments. These results indicate that Mpr1 can reduce intracellular ROS levels. It was also found that put2-disrupted yeast cells lacking Δ^1-pyrroline-5-carboxylate (P5C) dehydrogenease increased ROS levels and put2-disrupted cells that expressed MPR1 had considerably lower ROS levels. Mpr1 acetylates P5C or its equilibrium compound glutamtate-γ-semialdehyde. These results … More suggest that P5C is toxic to yeast cells due to the formation of ROS and that Mpr1 regulates the ROS level under P5C-induced oxidative stress.2) We found that yeast cells exhibited increased levels of ROS during freezing and thawing. Gene disruption and expression experiments indicated that Mpr1 protects yeast cells from freezing stress by reducing the intracellular levels of ROS. Similar results were obtained from ethanol stress. Similar results were obtained from ethanol stress. Gene disruption and expression experiments indicated that Mpr1 protect yeast cells from ethanol stress by reducing the intracellular ROS.3) We investigated whether MPR1 would function similarly in bacterial and plant cells and if its expression could confer resistance to AZC. Our studies confirm that MPR1 can function in a similar fashion in Escherichia coli and tobacco as in yeast to detoxify the toxic AZC so could potentially be used as a new selectable marker for E.coli and plant transformation.In summary, as a novel antioxidant mechanism, Mpr1 is supposed to reduce the intracellular oxidation level by acetylating the toxic metabolite(s) involved in ROS production, such as P5C/GSA. The proposed role of Mpr1 against oxidative stress could be promising for breeding novel stress-resistant yeast strains. Less

MPR1基因（Sigma 1278b基因用于盐酸盐抗性的基因）酿酒酵母odes N-乙酰基转移酶（MPR1）在这项研究中，将脯氨酸类似物Azetide-2-羧酸盐（AZC）解毒。 1）我们发现，MPR1破坏的细胞是高敏的毒性S，例如H_2O_2和热休克，并含有增加的活性氧（ROS）的水平。降低细胞内的ROS水平它的平衡化合物谷氨酸 - 甲醛。这些结果……更多地表明，由于ROS的形成，P5C对酵母细胞有毒，并且MPR1 ROS p5ced氧化应激。2）我们发现，在和融化过程中，我们发现了ROS S的增加。基因破坏和实验表明，MPR1通过降低ROS的细胞内水平来保护年度冻结，表达实验表明，Tatect酵母菌与细胞内ROS的Tatect酵母菌胁迫。3） IFS表达对AZC的耐药性。在P5C/GSA中，涉及ROS的有毒代谢物（S）的细胞内氧化作用，MPR1对氧化应激的支撑作用有望减少新型应激。

项目成果

Xing-Hai Zhang, H.Takagi, Jack.M.Widholm: "Expression of a novel yeast gene that detoxifies the proline analog azetidine-2-carboxylate confers resistance during tobacco seed germination, callus and shoot formation"Plant Cell Reports. Vol.22 No.8. 615-622

张星海、H.Takagi、Jack.M.Widholm：“表达一种新型酵母基因，可以解毒脯氨酸类似物氮杂环丁烷-2-羧酸酯，从而在烟草种子发芽、愈伤组织和芽形成过程中赋予抗性”《植物细胞报告》。

酵母に発見した「アセチル化酵素Mpr1」による新しい抗酸化メカニズム活性酸素種の生成に関わる物質を基質とした酸化ストレス防御システム

在酵母中发现的使用乙酰转移酶 Mpr1 的新抗氧化机制 使用参与活性氧生成的物质作为底物的氧化应激防御系统

• 发表时间: 2006
• 期刊: 化学と生物 44
• 作者: Yusei Miyamoto;Atsuhiro Kanayama;Jun-Ichiro Inoue;Yoshiko Sugita-Konishi Makoto Shimizu.;高木 博史
• 通讯作者: 高木 博史

Expression of a novel yeast gene that detoxifies the proline analog azetidine-2-carboxylate confers resistance during tobacco seed germination, callus and shoot formation

• DOI: 10.1007/s00299-003-0741-3
• 发表时间: 2004-03-01
• 期刊: PLANT CELL REPORTS
• 影响因子: 6.2
• 作者: Zhang, XH;Takagi, H;Widholm, JM
• 通讯作者: Widholm, JM

N-acetyltransferase Mpr1 confers freeze tolerance on Saccharomyces cerevisiae by reducing reactive oxygen species

• DOI: 10.1093/jb/mvi134
• 发表时间: 2005-10-01
• 期刊: JOURNAL OF BIOCHEMISTRY
• 影响因子: 2.7
• 作者: Du, XY;Takagi, H
• 通讯作者: Takagi, H

健康長寿研究成果報告書 酵母に見いだした酵素Mpr1よる新しい抗酸化メカニズム

健康长寿研究成果报告：酵母中发现Mpr1酶新抗氧化机制

• 发表时间: 2005
• 作者: M.Nomura;H.Takagi;高木 博史(分担執筆)
• 通讯作者: 高木 博史(分担執筆)