Role of RNA Editing by ADAR1 in Infection

ADAR1 RNA 编辑在感染中的作用

基本信息

批准号：
7413275
负责人：
JINGHUA YANG
金额：
$ 10.49万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-15 至 2010-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7413275
关键词：
2-5A Synthetase ADAR1 Acute Adenosine Adenoviruses Affect Amanitins Animals Area Award Bacteria Bacterial Pneumonia CXCL1 gene Cell Line Cell physiology Cells Chimera organism Collaborations DNA Microarray Chip DNA Microarray format Dactinomycin Disease Disease model Double-Stranded RNA Down-Regulation Endoribonucleases Endotoxemia Endotoxins Enzymes Genome Heterogeneous Nuclear RNA Host Defense Host Defense Mechanism Immune response Independent Scientist Award Infection Infectious Agent Inflammation Inflammatory Inflammatory Response Inosine Investigation Laboratories Lead Mediating Messenger RNA Mitogen-Activated Protein Kinases Molecular Mus Numbers Pancreatic ribonuclease Pathologic Pathway interactions Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Physiological Pneumonia Polymerase Production Productivity Protein Array Protein Biosynthesis Protein Kinase Protein Overexpression Proteins RNA Degradation RNA Editing RNA Processing RNA chemical synthesis Rate Research Reverse Transcriptase Polymerase Chain Reaction Ribonucleases Ribosomal RNA Role Scientist Staging Techniques Thymosin Time Toxin Transgenic Animals Transgenic Mice Translating Up-Regulation Virus adenosine deaminase cell growth eIF-2 Kinase mRNA Precursor mRNA Transcript Degradation macrophage mouse model prevent response skills

项目摘要

DESCRIPTION (provided by applicant): We have previously identified a cellular process that modifies pre-mRNA sequences via an RNA-specific adenosine deaminase or ADAR. Recently, we showed that ADAR1 is induced in macrophages by various infectious agents, including bacteria, viruses, and pathogenic toxins. In mice suffering from endotoxin-induced inflammation, we demonstrate that 5% of the adenosines found in mRNA are converted to inosine. We further show that up to 20% of adenosines in mRNA can be converted to inosine in cells transiently expressing ADAR1. With such high levels of inosine in mRNA, the translated proteins may differ dramatically from those encoded in the genome and the repertoire of protein production could be greatly amplified. RNA processing may also be substantially affected by the presence of inosine. Indeed, we found that the phosphorylation of elF2a is suppressed and the expression of dsRNA protein kinase (PKR) and 2',5'-oligoadenylate synthetase (2'5'OAS) is downregulated in cells that overexpress ADAR1. Therefore, we hypothesize that ADAR1 functions to sustain protein synthesis during infections, and antagonize inflammatory responses at certain stages of infection. Researches to confirm this hypothesis will lead me to disease-oriented studies in transgenic animals that are currently undeveloped in my laboratory. Receipt of the K02 award will allow me to devote 90% of my effort on the proposed research, meaningfully expand my collaborations with high quality scientists, and firmly establish the new lines of investigation to develop skills in these areas and to demonstrate increasing productivity. During award period, we plan to elucidate the physiological function of ADAR1 in modulating host response to infections. The first specific aim is to characterize the role of ADAR1 in disease models of pneumonia and endotoxemia in ADAR1 transgenic mice. Homozygous transgenic mice expressing an ADAR1-EGFP chimera will be produced. Endotoxin and adenovirus-induced acute inflammation will be established and characterized in these animals. The second specific aim proposes to elucidate the mechanism of ADARl-mediated RNA degradation in response to infectious agents. The last aim is to characterize the role of ADAR1 in modulating phosphorylation cascades in the mouse models of pneumonia and endotoxemia using protein array technique.

描述（由申请人提供）：我们先前已经确定了通过RNA特异性腺苷脱氨酶或ADAR修饰前MRNA序列的细胞过程。最近，我们表明ADAR1是由包括细菌，病毒和致病毒素在内的各种传染剂在巨噬细胞中诱导的。在患有内毒素诱导的炎症的小鼠中，我们证明了mRNA中发现的5％的腺苷转化为肌苷。我们进一步表明，mRNA中多达20％的腺苷可以转化为瞬时表达ADAR1的细胞中的插入。由于mRNA中的肌苷如此高，翻译的蛋白质可能与基因组中编码的蛋白质差异很大，并且蛋白质产生的曲目可能会大大扩增。 RNA处理也可能受到肌苷的存在。确实，我们发现ELF2A的磷酸化被抑制，DSRNA蛋白激酶（PKR）和2'，5'-寡胶质腺苷酸合成酶（2'5'OAS）的表达在过表达ADAR1的细胞中被下调。因此，我们假设ADAR1在感染过程中维持蛋白质的合成功能，并在某些感染阶段对抗炎症反应。确认这一假设的研究将使我对目前在我的实验室中尚未开发的转基因动物进行以疾病为导向的研究。获得K02奖的收到将使我能够将90％的努力投入到拟议的研究上，有意义地扩大了与高质量科学家的合作，并坚定建立了新的调查渠道，以在这些领域发展技能并证明生产力的提高。在奖励期间，我们计划阐明ADAR1在调节宿主对感染的反应中的生理功能。第一个具体目的是表征ADAR1在ADAR1转基因小鼠中肺炎和内毒素血症疾病模型中的作用。将产生表达ADAR1-EGFP嵌合体的纯合转基因小鼠。这些动物将建立和表征内毒素和腺病毒诱导的急性炎症。第二个特定目的是阐明响应传染剂的ADARL介导的RNA降解的机理。最后的目的是表征ADAR1在使用蛋白阵列技术的肺炎小鼠模型和内毒素毒素中调节磷酸化级联反应中的作用。