A Plant Protein Kinase and A Phosphatidylinositol 3-Kinase Involved in Vacuolar Protein Transport and Development

参与液泡蛋白转运和发育的植物蛋白激酶和磷脂酰肌醇 3-激酶

• 批准号: 9630108
• 负责人: Daryll Dewald
• 金额: $ 5.45万
• 依托单位: Utah State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-04-01 至 1999-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9630108&HistoricalAwards=false
• 关键词: Plant; Protein; Kinase; Phosphatidylinositol; Involved

96-30108 DeWald The plant cell vacuole is functionally similar to fungi vacuoles and to lysosome of mammalian cells. In addition, plant cell vacuoles have other functions; they function in regulating cytosolic pH, osmotic shock, and in the storage of proteins. Plant vacuole can occupy up to 90% of the total cellular volume. The maintenance of vacuoles requires the specific sorting of proteins and lipids from their sites of synthesis to the vacuoles. The process of recognition, sorting, and transport of proteins to the vacuole is complex. More than 30 genes have been implicated in vacuolar protein sorting. The goal of the proposed research is to determine the role of a phosphatidylinositol 3-kinase (PI 3- kinase) and that of a serine/threonine protein kinase in protein sorting to plant cell vacuole. The specific objectives are to study 1) the role of Vps 34p PI 3-kinase activity in protein sorting to plant vacuoles in Arabidopsis thaliana; 2) the role of Vsp 15p, a protein kinase, in protein sorting to plant vacuole in A. thaliana. The gene for Vps 15p will be cloned and sequence. Cloning of the gene will be done by functional complementation of a Vps 15p yeast mutant by transforming with an A. thaliana cDNA library, or by PCR-based amplification of gene fragments that will be used to probe the cDNA library to identify the gene. The role of Vps 15p and of Vps 34p will be assessed in A. thaliana by examining tissue and temporal gene expression patterns in wild type plants, and by documenting the effects of overexpressing the gene in transgenic plants. It is expected that this data will provide new insights into how protein transport is regulated in plants. %%% The plant cell vacuole is functionally similar to fungi vacuoles and to lysosome of mammalian cells. In addition, plant cell vacuoles have other functions; they function in regulating cytosolic pH, osmotic shock, and in the storage of proteins. Plant vacuole can occupy up to 90% of the total cellular vo lume. The maintenance of vacuoles requires the specific sorting of proteins and lipids from their sites of synthesis to the vacuoles. The process of recognition, sorting, and transport of proteins to the vacuole is complex. More than 30 genes have been implicated in vacuolar protein sorting. The goal of the proposed research is to determine the role of a phosphatidylinositol 3-kinase (PI 3- kinase) and that of a serine/threonine protein kinase in protein sorting to plant cell vacuole. The specific objectives are to study 1) the role of Vps 34p PI 3-kinase activity in protein sorting to plant vacuoles in Arabidopsis thaliana; 2) the role of Vsp 15p, a protein kinase, in protein sorting to plant vacuole in A. thaliana. The gene for Vps 15p will be cloned and sequence. Cloning of the gene will be done by functional complementation of a Vps 15p yeast mutant by transforming with an A. thaliana cDNA library, or by PCR-based amplification of gene fragments that will be used to probe the cDNA library to identify the gene. The role of Vps 15p and of Vps 34p will be assessed in A. thaliana by examining tissue and temporal gene expression patterns in wild type plants, and by documenting the effects of overexpressing the gene in transgenic plants. It is expected that this data will provide new insights into how protein transport is regulated in plants. ***

96-30108 DeWald 植物细胞液泡在功能上类似于真菌液泡和哺乳动物细胞的溶酶体。此外，植物细胞液泡还有其他功能；它们的功能是调节细胞质 p​​H 值、渗透压休克和蛋白质储存。植物液泡可占据细胞总体积的90%。液泡的维持需要对蛋白质和脂质从其合成位点到液泡进行特定的分类。蛋白质的识别、分类和运输到液泡的过程是复杂的。超过 30 个基因与液泡蛋白质分选有关。本研究的目的是确定磷脂酰肌醇 3-激酶（PI 3-激酶）和丝氨酸/苏氨酸蛋白激酶在植物细胞液泡蛋白质分选中的作用。具体目标是研究 1) Vps 34p PI 3-激酶活性在拟南芥植物液泡蛋白质分选中的作用； 2) Vsp 15p（一种蛋白激酶）在拟南芥植物液泡蛋白质分选中的作用。 Vps 15p 的基因将被克隆并测序。基因的克隆将通过用拟南芥 cDNA 文库转化对 Vps 15p 酵母突变体进行功能互补来完成，或者通过基于 PCR 的基因片段扩增来完成，该基因片段将用于探测 cDNA 文库以鉴定该基因。将通过检查野生型植物中的组织和时间基因表达模式以及记录转基因植物中过度表达该基因的影响来评估拟南芥中Vps 15p和Vps 34p的作用。预计这些数据将为植物中蛋白质运输的调控提供新的见解。 %%% 植物细胞液泡的功能类似于真菌液泡和哺乳动物细胞的溶酶体。此外，植物细胞液泡还有其他功能；它们的功能是调节细胞质 p​​H 值、渗透压休克和蛋白质储存。植物液泡可占细胞总体积的90%。液泡的维持需要对蛋白质和脂质从其合成位点到液泡进行特定的分类。蛋白质的识别、分类和运输到液泡的过程是复杂的。超过 30 个基因与液泡蛋白质分选有关。本研究的目的是确定磷脂酰肌醇 3-激酶（PI 3-激酶）和丝氨酸/苏氨酸蛋白激酶在植物细胞液泡蛋白质分选中的作用。具体目标是研究 1) Vps 34p PI 3-激酶活性在拟南芥植物液泡蛋白质分选中的作用； 2) Vsp 15p（一种蛋白激酶）在拟南芥植物液泡蛋白质分选中的作用。 Vps 15p 的基因将被克隆并测序。基因的克隆将通过用拟南芥 cDNA 文库转化对 Vps 15p 酵母突变体进行功能互补来完成，或者通过基于 PCR 的基因片段扩增来完成，该基因片段将用于探测 cDNA 文库以鉴定该基因。将通过检查野生型植物中的组织和时间基因表达模式以及记录转基因植物中过度表达该基因的影响来评估拟南芥中Vps 15p和Vps 34p的作用。预计这些数据将为植物中蛋白质运输的调控提供新的见解。 ***