Protein-protein interactions in mono- and bi-partite geminiviruses: functional analysis and use of aptamers for plant protection

本提案代表了我们对双病毒科植物ssDNA病毒(番茄黄卷叶病毒（TYLCV）、西瓜绿僵病毒（WmCSV）和印度木薯花叶病毒（ICMV）的感染、传播和致病性的分子成分和过程的研究、鉴定和表征的继续。获得的信息将用于开发新的策略，使植物对双病毒感染具有抗性。这将通过在植物中表达抑制肽来实现，抑制肽会干扰双病毒蛋白在细胞间和细胞内的运动。随机肽库和酵母双杂交（Hoppe-Seyler et al., 2004）以及反向双杂交（Barr et al., 2004）系统将用于选择此类抑制肽。事实上，我们已经成功地利用Y2H系统在培养细胞中选择了一种抑制病毒复制（hiv - 1）的肽（Armon-Omer等人，2007）。同样的策略将被我们在目前的项目工程中使用，通过表达显性干扰肽（适体）的抗病毒转基因植物最近被描述为一种新的和可行的方法，以控制RNA病毒感染(Rudolph等/。（2003年）以及双病毒的复制（Lopez-Ochoa等人，2006年）。在这里，我们建议利用我们在随机肽库的使用和双病毒蛋白相互作用方面的知识来产生抑制肽，以获得病毒抗性植物。

This proposal represents a continuation of our ongoing efforts to study, identify and characterize the molecular components and the processes involved in infection, spread and pathogenicity of plant ssDNA viruses from the Geminiviridae family: Tomato yellow leaf curl virus (TYLCV), Watermelone chlorotic stunt virus (WmCSV), and Indian cassava mosaic virus (ICMV). The information gained will be used to develop novel strategies to render plants resistant to geminivirus infection. This will be achieved by expressing inhibitory peptides in planta that will interfere with inter- and intracellular movement of geminiviral proteins. A random peptide library and the yeast two hybrid (Hoppe-Seyler et al., 2004) as well as the reverse two-hybrid (Barr et al., 2004) systems will be used to select such inhibitory peptides. Indeed, we have already succeeded, using Y2H system in selecting a peptide that inhibits virus replication (HIV-l) in cultured cells (Armon-Omer et al., 2007). The same strategy will be used by us in the present project Engineering of virus-resistant transgenic plants by expressing dominant interfering peptides (aptamers) was described recently as a novel and feasible approach in order to control an RNA virus infection (Rudolph et a/., 2003) as well as the replication of a geminivirus (Lopez-Ochoa et aly 2006). Here, we propose to exploit our knowledge on the use of random peptide libraries and on the interactions of geminivirus proteins to generate inhibitory peptides to obtain virus resistant plants.