AGROBACTERIUM VIRD2 INTERACTION WITH HOST CYCLOPHILINS

农杆菌 VIRD2 与宿主亲环蛋白的相互作用

• 批准号: 6178841
• 负责人: DEREK W WOOD
• 金额: $ 3.92万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-08-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6178841
• 关键词: Agrobacterium tumefaciens; Arabidopsis; affinity chromatography; antisense nucleic acid; bacteria infection mechanism; bacterial proteins; cyclosporines; genetic library; host organism interaction; immunoprecipitation; intracellular transport; peptidylprolyl isomerase; polymerase chain reaction; yeast two hybrid system

The long term objective of this proposal is to clarify the cellular role of cyclophilins. Cyclophilins are present in a wide range of cell types suggesting that they play an essential role in cell biology. Cyclophilins have recently been implicated in a number of processes including apoptosis, signal transduction, protein stabilization (chaperones), immunosuppression and pathogenesis of a variety of organisms (i.e. Leishmania and HIV-1). The specific goals of this research are to: 1) verify the interaction between the VirD2 protein of Agobacterium tumefaciens and two cyclophilins of Arabidopsis thaliana, 2) demonstrate that this interaction is important in tumorigenesis, 3) examine the interaction of these cyclophilins with VirD2 and other host proteins in an effort to define the cellular role of host cyclophilins and 4) investigate the involvement of cyclophilins in Agrobacterium-mediated transformation of yeast. Affinity chromatography and in vivo across- linking analysis will confirm the specific interactions between VirD2 and cyclophilins previously defined using yeast two-hybrid analysis. Cyclosporin A inhibition antisense RNA analysis will be used to examine the importance of this interaction in tumorigenesis initiated by agrobacterium. Site and region specific mutagenesis will be used in concert with forward and reverse yeast two-hybrid analysis to identify VirD2 residues essential to this interaction and to construct suppressor mutations in cyclophilins will be identified in an effort to elucidate host transport or signal transduction pathways linked to cyclophilins. Further work will utilize yeast as a tractable genetic model in which to examine the effect of cyclophilin disruptions on transformations.

该提议的长期目标是阐明细胞作用 环蛋白。环胞素存在于多种细胞类型中 表明它们在细胞生物学中起着至关重要的作用。环胞素 最近与许多过程有关 凋亡，信号转导，蛋白质稳定（伴侣）， 各种生物的免疫抑制和发病机理（即 利什曼尼亚和HIV-1）。这项研究的具体目标是：1） 验证Agobacterium的Vird2蛋白之间的相互作用 Tumefaciens和拟南芥的两个环友，2）证明 这种相互作用在肿瘤发生中很重要，3）检查 这些环粒蛋白与VIRD2和其他宿主蛋白的相互作用 努力定义宿主环蛋白的细胞作用和4） 研究环胞素参与农杆菌介导的 酵母的转化。亲和色谱和体内 链接分析将确认VIRD2和 先前使用酵母双杂交分析定义的环胞素。 环孢菌素A抑制反义RNA分析将用于检查 这种相互作用在肿瘤发生中的重要性 农杆菌。位点和区域特异性诱变将用于 与前进和反向酵母双杂交分析进行音乐会，以识别 VIRD2残基对这种相互作用必不可少的并构造抑制剂 为了阐明，环胞素的突变将被确定 宿主运输或信号转导途径与环胞素相关。 进一步的工作将利用酵母作为一种可进行的遗传模型 检查环磷脂破坏对转化的影响。