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: 9030945
• 负责人: Hailing Jin
• 金额: $ 31.02万
• 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/9030945
• 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; Coupled; Detection; Eukaryota; Fluorescence; Funding; Gene Silencing; Genes; Goals; Health; Human; Immune response; Immune system; Immunity; Infection; 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; disorder control; helicase; in vitro Assay; 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（由适用提供）：包括动物和植物在内的真核生物已经发展出复杂的免疫学系统，以保护自己免受病原体的侵害。小RNA（SRNA）是短而非编码的调节RNA，通过与Argonaute（AGO）蛋白结合并用完整序列引导基因沉默来影响RNA干扰（RNAI）。我们的工作和其他人的工作表明，源自宿主或病原体的SRNA通过AGO蛋白质发挥作用，并在宿主 - 病原体相互作用中起重要作用。但是，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史的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，我们假设它们是通过其相关蛋白来修饰和调节的，以调节SRNA介导的基因沉默。在这里，我们试图通过使用模型的植物病原体相互作用系统 - 拟南芥及其细菌病原体假单胞菌和促进性肌no虫和侵略性的真实性真菌病原病原体，旨在了解RNAi途径中AGO的修改和调节，以及宿主抗菌和抗真菌免疫复杂的修改和调节。在最后一个资金期间，我们已经确定了调节宿主免疫的宿主SRNA的潜水员类别，并证明拟南芥AGO2（atago2）高度诱导了细菌感染，并对宿主免疫学进行了积极调节。我们还表明，B. cinerea将其SRNA传递到宿主细胞中，劫持宿主Atago1以抑制宿主免疫学基因成功感染。为了获得对Atago1和Atago2如何调节的机械理解，我们通过蛋白质组学分析确定了它们相关的蛋白质。这种更新将研究Atago1和Atago2对RNAi途径中的相关蛋白和宿主先天免疫史的修饰和调节。这项研究的输出将大大提高我们对宿主免疫反应中RNAi的动态控制的理解，这将对农业和人类健康产生重大影响。具体目的是：1。阐明蛋白精氨酸甲基转移酶在宿主免疫中Atago功能的修饰和调节中的作用。 2。研究宿主免疫中DDX3样RNA解旋酶在调节Atago功能中的功能。 3。研究蛋白磷酸酶在调节阿特哥和宿主免疫中的作用。