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Research PGR: Genome-enabled characterization of orphan receptor-like kinases in plants.

研究 PGR：植物中孤儿受体样激酶的基因组表征。

基本信息

批准号：
2048410
负责人：
Laura Bartley
金额：
$ 200万
依托单位：
Washington State University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-05-31
项目状态：
未结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2048410&HistoricalAwards=false
关键词：
Research PGR Genome enabled characterization

项目摘要

This project will improve understanding of an important class of proteins, the so-called receptor-like kinases. Like cellular sentries, receptor-like kinases have crucial functions in sensing and orchestrating responses to environmental cues. Kinases perceive molecular signals from other organisms (like pathogens) or from neighboring cells and pass signals along to other proteins to initiate defense responses or other changes, such as halting growth. Consistent with the importance of these functions, kinases are one of the most abundant plant protein families; however, kinases have also proven to be difficult to study. New advances in both understanding and technology will be used in this project to enable matching of plant kinases to the signals that they detect and to the downstream “recipients” of the information, thereby revealing the function of these proteins. Improvements in knowledge of kinases will open new avenues for improving plant production for food, feed, fiber, and energy. In addition, the project will train at least five young scientists to use advanced technologies and will provide plant science educational opportunities to ~60 teachers via professional development workshops, introduce ~30 Native American high school and undergraduate students to research via week-long plant signaling research project workshops, and provide ~40 summer and academic year undergraduate research opportunities that meld plant and data science.This research will characterize the functions and substrates of receptor-like kinases (RLKs) of Arabidopsis, rice, and soybean. Objective 1 aims to reveal RLK functions by combing quantification of phosphorylation and S-acylation (e.g., palmitoylation) as indicators of involvement of an RLK in a biological process. This analysis will take advantage of new results that suggest that these posttranslational modifications are antagonistic. The project will use proteomics technology to deeply and quantitatively sample total and plasma membrane-enriched proteins during rice internode development, with and without drought stress; and soybean and rice root responses to conserved putative small secreted peptides. The relatively well-understood Arabidopsis- Pseudomonas syringae interaction will be examined to optimize methods. The focus on peptide hormone ligands and root growth will survey broadly across predicted small secreted peptide families. Objective 2 takes a complementary approach to revealing RLK function, by creating deep (12k) peptide libraries for each species and screening them against purified protein kinases with the goal of identifying kinase clients for at least 25 novel kinases. The kinase client assay accelerates assignment of downstream signaling and its usefulness has been demonstrated by rapid community adoption. In Objective 3, experimental results will be extended using machine learning approaches to predict plant phosphorylation sites and specific kinase clients. Proteome data will be used for molecular network reconstruction and for comparative analysis to identify patterns in RLK signaling. Results and predictions will be used to enhance previously established databases, P3DB and SoyKB, which are already well-used by the community, amplifying project value.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这个项目将提高对一类重要蛋白质的理解，即所谓的受体样激酶。与细胞哨兵一样，受体样激酶在感知和协调对环境提示的反应方面具有至关重要的功能。激酶可以感知来自其他生物体(如病原体)或邻近细胞的分子信号，并将信号传递给其他蛋白质，以启动防御反应或其他变化，如停止生长。与这些功能的重要性一致的是，激酶是最丰富的植物蛋白家族之一；然而，激酶也被证明是很难研究的。该项目将利用理解和技术方面的新进展，使植物激酶与它们检测到的信号和信息的下游“接受者”相匹配，从而揭示这些蛋白质的功能。对激酶知识的改进将为提高植物的食物、饲料、纤维和能源产量开辟新的途径。此外，该项目将培训至少5名年轻科学家使用先进技术，并将通过专业发展研讨会为约60名教师提供植物科学教育机会，通过为期一周的植物信号研究项目研讨会介绍约30名美国原住民高中生和本科生进行研究，并提供约40个融合植物和数据科学的夏季和学年本科生研究机会。这项研究将表征拟南芥、水稻和大豆受体样蛋白(RLK)的功能和底物。目的1结合磷酸化和S酰化(如棕榈酰化)作为RLK参与生物过程的指标，揭示RLK的功能。这一分析将利用新的结果，这些结果表明这些翻译后修饰是对抗性的。该项目将利用蛋白质组学技术，深入和定量地采样水稻节间发育过程中总的和质膜丰富的蛋白质，包括干旱胁迫和不干旱胁迫；以及大豆和水稻根系对保守的假定分泌小肽的反应。将对相对较好地理解的拟南芥-丁香假单胞菌相互作用进行研究，以优化方法。对多肽激素配体和根生长的关注将在预测的小分泌肽家族中进行广泛的调查。目的2采用一种补充的方法来揭示RLK的功能，通过为每个物种创建深层(12K)多肽文库，并以纯化的蛋白激酶为对照进行筛选，目的是识别至少25种新的激酶的激酶客户。激酶客户端分析加速了下游信号的分配，其有效性已被迅速的社区采用所证明。在目标3中，将使用机器学习方法来扩展实验结果，以预测植物磷酸化位点和特定的激酶客户。蛋白质组数据将用于分子网络重建和比较分析，以确定RLK信号的模式。结果和预测将用于增强以前建立的数据库，P3DB和SoyKB，这些数据库已经被社区很好地使用，放大项目价值。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。