Activation of plastid-localized latent polyphenol oxidase by autophagy induced by sucrose-starvation
蔗糖饥饿诱导的自噬激活质体定位的潜在多酚氧化酶
基本信息
- 批准号:12640630
- 负责人:
- 金额:$ 2.05万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2000
- 资助国家:日本
- 起止时间:2000 至 2002
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Polyphenol oxidase (PPO) that is translated as precursor protein in cytoplasm, then transported and localized in plastids as a latent form. However, PPO's substrates are localized in the other compartment (vacuoles). Plant PPO is well known as one of the most popular enzymes that are involved in browning of the injured or diseased tissues. However, its biological function in the living plant cell is not clear. In the present study, we demonstrate that the plastid-localized latent PPO is most probably activated by proteolytic cleavage involved in autophagy. Main experimental results are shown as follows.1) We cloned 2 PPO isoform cDNAs from sweet potato cells.2) Expression of PPO was induced by cell transfer to fresh medium, cell proliferation and treatment with methyl jasmonate.3) Latent PPO was activated by proteolytic cleavage in the cells by sucrose starvation or methyl jasmonate.4) A vacuole-localized cysteine endopeptidase (PPOase) that can cleave the latent PPO was partially purified. PPOase showed a high substrate specificity for PPO.5) Proteolytic cleavage of latent PPO in the sweet potato cells that had been cultured in sucrose-deleted medium was markedly inhibited by E-64 or 3-methyladenine. Plastid number severely decrease by sucrose starvation, but both E-64 and 3-methyladenine also inhibited the breakdown of plastids.6) Vacuoles were isolated from E-64 treated cells that had been cultured in sucrose-deleted medium. The vacuoles containing many DAPI-stained structures that seemed to be plastids were observed under fluorescent microscopy.These experimental results indicate that autophagy is induced by sucrose starvation in sweet potato cells, and the active PPO is generated by proteolytic cleavage of the plastid-localized latent PPO involved in cysteine endopeptidase in the vacuoles.
多酚氧化酶(PPO)是一种在细胞质中作为前体蛋白翻译,然后以潜伏形式运输并定位于质体中的酶。然而,PPO的底物位于另一个隔室(液泡)中。植物多酚氧化酶(PPO)是参与植物损伤或病变组织的布朗宁的最常见的酶之一。然而,其在植物活细胞中的生物学功能尚不清楚。在本研究中,我们证明了质体定位的潜在PPO最有可能被激活的蛋白水解裂解参与自噬。主要实验结果如下:1)从甘薯细胞中克隆了2个PPO同工型cDNA。2)通过细胞转移到新鲜培养基中诱导PPO表达,细胞增殖和茉莉酸甲酯处理; 3)蔗糖饥饿或茉莉酸甲酯引起的细胞中的蛋白水解裂解激活了潜在的PPO; 4)液泡定位的半胱氨酸内肽酶(PPO酶)部分纯化了能切割潜在PPO的酶。5)E-64和3-甲基腺嘌呤能显著抑制甘薯细胞中潜在PPO的蛋白酶解。蔗糖饥饿使质体数目严重减少,但E-64和3-甲基腺嘌呤也能抑制质体的破裂。6)从E-64处理的细胞中分离出了异源醇。荧光显微镜下观察到液泡中含有许多DAPI染色的类似质体的结构,这些实验结果表明,蔗糖饥饿诱导甘薯细胞发生自噬,而活性PPO是由液泡中半胱氨酸内肽酶参与的位于质体中的潜在PPO被蛋白水解裂解而产生的。
项目成果
期刊论文数量(21)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Xu, W., Shioiri, H., Kojima, M., Nozue, M.: "Primarys structure and expression of a 24-kD vacuolar protein(VP24)precursor in anthocyanin-producing cells of sweet potato in suspension culture"Plant Physiology. 125. 447-455 (2001)
Xu, W., Shioiri, H., Kojima, M., Nozue, M.:“悬浮培养的甘薯花青素生产细胞中 24-kD 液泡蛋白 (VP24) 前体的初级结构和表达”植物生理学
- DOI:
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- 影响因子:0
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Xu, W., Shioiri, H., Kojima, M. and Nozue, M.: "Primary structure and expression of a 24-kD vacuolar protein (VP24) precursor in anthocyanin-producing cells of sweet potato in suspension culture."Plant Physiology. 125. 447-455 (2001)
Xu, W.、Shioiri, H.、Kojima, M. 和 Nozue, M.:“悬浮培养的甘薯花青素生成细胞中 24-kD 液泡蛋白 (VP24) 前体的一级结构和表达。”
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- 影响因子:0
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Xu,W.,Moriya,K.,Yamada,K.,Nishimura,M.,Shioiri,H.,Kojima,M.,Nozue,M.: "Detection and characterization of a 36-kDa peptide in C-terminal region of a 24-kDa vacuolar protein (VP24) precursor in anthocyanin-producing sweet potato cells in suspension culture"
Xu,W.,Moriya,K.,Yamada,K.,Nishimura,M.,Shioiri,H.,Kojima,M.,Nozue,M.:“C 末端区域 36-kDa 肽的检测和表征
- DOI:
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- 影响因子:0
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Xu, W., Moriya, K., Yamada, K., Mishimura, M., Shioiri, H., Kojima, M., Nozue, M.: "Detection and characterization of a 36-kDa peptide in C-terminal region of a 24-kDa vacuolar protein (VP24) precursor in anthocyanin-producing sweet potato cells in suspen
Xu, W.、Moriya, K.、Yamada, K.、Mishimura, M.、Shioiri, H.、Kojima, M.、Nozue, M.:“C 末端区域 36 kDa 肽的检测和表征
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- 影响因子:0
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Xu, W., Shioiri, H., Kojima, M. and Nozue, M.: "Expression and localization of a 36-kDa peptide derived from a 24-kDa vacuolar protein (VP24) precursor in anthocyanin-producing sweet potato cells in suspension culture."Plant Biotechnology. 18. 203-208 (20
Xu, W.、Shioiri, H.、Kojima, M. 和 Nozue, M.:“源自 24-kDa 液泡蛋白 (VP24) 前体的 36-kDa 肽在产生花青素的甘薯细胞中的表达和定位
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24580367 - 财政年份:2012
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