Modification and dynamic regulation of Argonaute proteins-the key small RNA pathway components in host innate immunity

Argonaute蛋白的修饰和动态调控——宿主先天免疫中关键的小RNA通路成分

• 批准号: 9231457
• 负责人: Hailing Jin
• 金额: $ 31.06万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9231457
• 关键词: Agriculture; Animal Model; Animals; Anti-Bacterial Agents; Antibodies; Antifungal Agents; Arabidopsis; Arginine; Bacterial Infections; Binding; Biological Assay; Botrytis; Catalytic Domain; Cells; Co-Immunoprecipitations; Complement; Coupled; Detection; Eukaryota; Fluorescence; Funding; Gene Silencing; Genes; Goals; Health; Human; Immune response; Immune system; Immunity; In Vitro; Infection; Innate Immune System; Invertebrates; Knock-out; Mammals; Mass Spectrum Analysis; Mediating; Methylation; Modification; Mouse-ear Cress; Mutation; Natural Immunity; Organism; Output; Pathway interactions; Phosphoric Monoester Hydrolases; Phosphorylation Site; Plant Model; Plants; Play; Protein Family; Protein Phosphatase 2A Regulatory Subunit PR53; Protein phosphatase; Protein-Arginine N-Methyltransferase; Proteins; Proteomics; Pseudomonas syringae; RNA Helicase; RNA Interference; RNA Interference Pathway; Regulation; Research; Role; Site; Small RNA; System; Testing; United States National Institutes of Health; Untranslated RNA; Viral; Virulence; Work; dimethylarginine; disorder control; helicase; in vivo; mutant; novel; pathogen; protein phosphatase 2C; public health relevance

 DESCRIPTION (provided by applicant): Eukaryotic organisms, including animals and plants, have evolved sophisticated immune systems to protect themselves against pathogens. Small RNAs (sRNAs) are short, non-coding regulatory RNAs that induce RNA interference (RNAi) by binding to Argonaute (AGO) proteins and directing silencing of genes with complementary sequences. Our work and those of others have demonstrated that sRNAs, derived from either hosts or pathogens, function through AGO proteins and play an important role in host-pathogen interactions. However, the underlying regulatory mechanisms remain elusive. Our long-term goal is to understand the dynamic regulation of sRNAs and RNAi in host innate immunity. AGO proteins are key players of sRNA- mediated gene silencing, and we hypothesize that they are modified and regulated by their associated proteins to modulate sRNA-mediated gene silencing. Here, we seek to understand the modification and regulation of AGOs in RNAi pathway, and in host antibacterial and antifungal immune responses, by using model plant-pathogen interaction systems - Arabidopsis thaliana and its bacterial pathogen Pseudomonas syringae and aggressive fungal pathogen Botrytis cinerea. During the last funding period, we have identified diverse classes of host sRNAs regulating host immunity, and have demonstrated that Arabidopsis AGO2 (AtAGO2) is highly induced by bacterial infection and positively regulates host immunity. We have also shown that B. cinerea delivers its sRNAs into host cells and hijacks host AtAGO1 to suppress host immunity genes for successful infection. To obtain a mechanistic understanding of how AtAGO1 and AtAGO2 are regulated, we have identified their associated proteins via proteomics analysis. This renewal will study the modification and regulation of AtAGO1 and AtAGO2 by their associated proteins in RNAi pathways and host innate immunity. The output of this study will significantly advance our understanding of dynamic control of RNAi in host immune responses against pathogens, which will have great impact on both agricultural and human health. The specific aims are: 1. Elucidate the role of protein arginine methyltransferases in modification and regulation of AtAGO function in host immunity. 2. Investigate the function of DDX3-like RNA helicases in modulating AtAGO function in host immunity. 3. Study the role of protein phosphatases in modulating AtAGO and host immunity.

 描述（由申请人提供）：真核生物，包括动物和植物，已经进化出复杂的免疫系统来保护自己免受病原体侵害。小 RNA (sRNA) 是短的非编码调节 RNA，可通过与 Argonaute (AGO) 蛋白结合并指导具有互补序列的基因沉默来诱导 RNA 干扰 (RNAi)。我们和其他人的工作已经证明，源自宿主或病原体的 sRNA 通过 AGO 蛋白发挥作用，并在宿主-病原体相互作用中发挥重要作用。然而，潜在的监管机制仍然难以捉摸。我们的长期目标是了解宿主先天免疫中 sRNA 和 RNAi 的动态调节。 AGO 蛋白是 sRNA 介导的基因沉默的关键参与者，我们假设它们被相关蛋白修饰和调节，以调节 sRNA 介导的基因沉默。在这里，我们试图通过使用模型植物-病原体相互作用系统——拟南芥及其细菌病原体丁香假单胞菌和侵袭性真菌病原体灰葡萄孢，了解 AGO 在 RNAi 途径以及宿主抗菌和抗真菌免疫反应中的修饰和调节。在上一个资助期间，我们已经鉴定了多种调节宿主免疫的宿主sRNA，并证明拟南芥AGO2（AtAGO2）是由细菌感染高度诱导的，并且正向调节宿主免疫。我们还发现，灰霉病菌将其 sRNA 传递到宿主细胞中，并劫持宿主 AtAGO1，以抑制宿主免疫基因，从而成功感染。为了获得对 AtAGO1 和 AtAGO2 如何调控的机制了解，我们通过蛋白质组学分析鉴定了它们的相关蛋白。这项更新将研究 AtAGO1 和 AtAGO2 的相关蛋白在 RNAi 途径和宿主先天免疫中的修饰和调节。这项研究的成果将显着增进我们对RNAi在宿主针对病原体的免疫反应中的动态控制的理解，这将对农业和人类健康产生巨大影响。具体目标是： 1. 阐明蛋白质精氨酸甲基转移酶在修饰和调节宿主免疫中 AtAGO 功能中的作用。 2. 研究DDX3样RNA解旋酶在调节宿主免疫中AtAGO功能中的功能。 3. 研究蛋白磷酸酶在调节AtAGO和宿主免疫中的作用。