Elucidating RIN4_Mediated Immune Signaling Cascades in Arabidopsis

阐明拟南芥中 RIN4_介导的免疫信号级联

• 批准号: 8817025
• 负责人: Gitta Laurel Coaker
• 金额: $ 32.97万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8817025
• 关键词: ATP phosphohydrolase; Affect; Agriculture; Animals; Antigen-Antibody Complex; Arabidopsis; Architecture; Binding; Biochemical; Biological; Cells; Client; Co-Immunoprecipitations; Complement; Complex; Comprehension; Coupled; Development; Disease; Disease Resistance; Environment; Epitopes; Eukaryota; Event; Evolution; Flagellin; Genetic; Genotype; Hand; Health; Human; Immune; Immune response; Immune system; Immunologic Receptors; In Vitro; Infection; Investigation; Laboratories; Leucine-Rich Repeat; Link; MAP Kinase Gene; Mass Spectrum Analysis; Mediating; Membrane; Modeling; Modification; Molecular; Monitor; Mouse-ear Cress; Mustard; Mutation; Natural Immunity; Nucleotides; Organism; Output; Perception; Phosphorylation; Plant Proteins; Plants; Post-Translational Protein Processing; Principal Investigator; Productivity; Protein Family; Proteins; Pseudomonas; Pseudomonas syringae; Receptor Activation; Receptor Signaling; Recombinants; Relative (related person); Research; Resistance; Rest; Role; Signal Transduction; Signaling Protein; System; Testing; Transgenic Plants; Validation; Vertebrates; Virulence; Widespread Disease; adapter protein; disease phenotype; disorder control; food security; genetic analysis; immune activation; improved; innovation; interest; leucine-rich repeat protein; member; microbial; mutant; novel; pathogen; preference; programs; protein complex; public health relevance; receptor; reconstitution; research study; stoichiometry; tool; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): The innate, or germline encoded, immune system is an ancient and evolutionary conserved defense system mediating pathogen recognition in humans, animals, and plants. Some innate immune receptors recognize conserved microbial features. Scientific studies have revealed that similar microbial features are recognized across diverse hosts as a result of convergent evolution. For example, in animals, humans and plants bacterial flagellin is recognized by leucine-rich repeat receptors leading to the downstream activation of MAPK cascades and activation of disease resistance. Intracellular immune receptors possessing nucleotide binding, leucine- rich repeat domains (NLRs) act to recognize pathogen effector proteins delivered into plant cells. In humans and animals, NLR receptors can recognize microbial features and, in some cases, effectors. Despite the involvement of analogous immune receptors across diverse organisms, the molecular mechanisms controlling receptor activation and interconnected downstream signaling nodes remain elusive. This is likely due to functional redundancy and lethality underlying key signaling sectors. In this application, we seek to understand the early events underlying NLR receptor activation in plants using the mustard relative, Arabidopsis thaliana, and the bacterial pathogen Pseudomonas syringae as a model. The application will focus on RIN4, a conserved plant protein that can regulate aspects of perception of microbial features, can interact with the NLR receptors RPM1 and RPS2, and is targeted by multiple pathogen effectors. We are able to purify biologically active RPM1 and associated proteins, which will be used to biochemically, investigate NLR receptor activation in vitro. The RPM1 complex will be isolated from Arabidopsis at a resting and activated state and associated proteins will be identified by mass spectrometry. We have optimized conditions for RPM1 complex purification and have several interesting signaling proteins in hand for downstream biochemical, cell biological and genetic validation experiments. The molecular mechanisms controlling effector-induced RIN4 phosphorylation and cleavage for promoting pathogen virulence in susceptible genotypes will be investigated. A greater understanding of how plant immune receptors recognize pathogens will fundamentally advance our understanding of receptor recognition across diverse organisms and is required for the development of novel, environmentally friendly disease control strategies. The specific aims of the project are: 1. Identify changes in complex assembly, enzymatic activity, and known protein associations upon activation of the immune receptor RPM1 in vitro. 2. Identify and characterize NLR protein complexes in a resting and activated state. 3. Investigate the importance of effector-induced RIN4 modifications for interaction with host proteins.

描述（由申请人提供）：先天或种系编码免疫系统是人类、动物和植物中介导病原体识别的古老且进化保守的防御系统。一些先天免疫受体识别保守的微生物特征。科学研究表明，作为趋同进化的结果，相似的微生物特征在不同宿主中得到认可。例如，在动物、人类和植物中，细菌鞭毛蛋白被富含亮氨酸的重复受体识别，导致MAPK级联的下游激活和抗病性的激活。具有核苷酸结合、富含亮氨酸重复结构域（NLRs）的细胞内免疫受体可识别进入植物细胞的病原体效应蛋白。在人类和动物中，NLR受体可以识别微生物特征，在某些情况下，还可以识别效应器。尽管在不同的生物体中有类似的免疫受体参与，但控制受体激活和相互连接的下游信号节点的分子机制仍然难以捉摸。这可能是由于关键信号部门的功能冗余和致命性。在这个应用中，我们试图了解NLR受体在植物中激活的早期事件，使用芥菜的近亲拟南芥和细菌病原体丁香假单胞菌作为模型。该申请将重点关注RIN4，这是一种保守的植物蛋白，可以调节微生物特征的感知方面，可以与NLR受体RPM1和RPS2相互作用，并且是多种病原体效应物的靶标。我们能够纯化具有生物活性的RPM1和相关蛋白，这些蛋白将用于体外生化研究NLR受体的激活。RPM1复合体将从静止和激活状态的拟南芥中分离出来，并通过质谱法鉴定相关蛋白。我们已经优化了RPM1复合体的纯化条件，并有几个有趣的信号蛋白在手中的下游生化，细胞生物学和遗传验证实验。在敏感基因型中，控制效应物诱导的RIN4磷酸化和裂解以促进病原体毒力的分子机制将被研究。更好地了解植物免疫受体如何识别病原体将从根本上推进我们对不同生物体中受体识别的理解，并且需要开发新的环境友好型疾病控制策略。该项目的具体目标是：1。鉴定体外免疫受体RPM1激活后复合物组装、酶活性和已知蛋白关联的变化。2. 鉴定和表征静息和激活状态下的NLR蛋白复合物。3. 研究效应剂诱导的RIN4修饰对与宿主蛋白相互作用的重要性。