Prediction and Validation of Phospho-Regulatory Sites in Crop Plant Proteomes

作物蛋白质组中磷酸调节位点的预测和验证

• 批准号: 0321450
• 负责人: Jeffrey Harper
• 金额: --
• 依托单位: The Scripps Research Institute
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0321450&HistoricalAwards=false
• 关键词: Prediction; Validation; Phospho; Regulatory; Sites

The genomes of vascular plants encode more than 1000 protein kinases. The long-term goal is todefine all the regulatory phosphorylation sites in crop plant proteomes. Such sites represent potential control points for all aspects of plant growth, including responses to biotic and abiotic stress. The focus is to use bioinformatics to predict a subset of potentially important sites of phospho-regulation in the rice proteome, and then use a peptide-based experimental strategy to start validating those predictions. The four Specific Aims are:1. Improve bioinformatics tools to identify motifs involved in phospho-signaling in plants.2. Conduct a clustering analysis of all Ser and Thr containing sub-sequences in plant proteomes.3. Design peptide array platform to profile kinase activities in vitro.4. Identify peptides that can disrupt in vivo signaling pathways (more than 50 will be tested).The uniqueness and strength of the proposed research rests upon the underlying rationale that acomputational comparison of two plant proteomes can be used as a 'noise filter' to identifyimportant sequence motifs that are fundamental to signaling pathways conserved between monocots and dicots. This filtering provides the critical foundation for implementing 'peptide-disruptors' as a powerful new approach to understanding and manipulating the dynamics of the plant cell phosphorylome. DNA constructs for all genes found to encode a 'peptide disruptor' or function as a reporter for a particular kinase activity will be deposited at the ABRC at Ohio State University, and will be freely available.

维管植物的基因组编码 1000 多种蛋白激酶。长期目标是定义作物蛋白质组中的所有调节磷酸化位点。这些位点代表了植物生长各个方面的潜在控制点，包括对生物和非生物胁迫的反应。重点是使用生物信息学来预测水稻蛋白质组中磷酸调节的潜在重要位点的子集，然后使用基于肽的实验策略开始验证这些预测。这四个具体目标是： 1．改进生物信息学工具来识别植物中参与磷酸信号传导的基序。2．对植物蛋白质组中所有含有Ser和Thr的子序列进行聚类分析。3．设计肽阵列平台以分析体外激酶活性。4．鉴定可以破坏体内信号通路的肽（将测试超过 50 种）。拟议研究的独特性和强度取决于以下基本原理：两种植物蛋白质组的计算比较可以用作“噪声过滤器”，以识别对于单子叶植物和双子叶植物之间保守的信号通路至关重要的重要序列基序。这种过滤为实现“肽干扰物”作为理解和操纵植物细胞磷酰基动态的强大新方法提供了关键基础。所有被发现编码“肽干扰物”或作为特定激酶活性报告基因的 DNA 构建体将存放在俄亥俄州立大学的 ABRC，并且将免费提供。