Study the roles of RNA editing enzyme ADAR1 in the regulation of cell death

研究RNA编辑酶ADAR1在细胞死亡调控中的作用

• 批准号: 7737674
• 负责人: Qingde Wang
• 金额: $ 18.63万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-05 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7737674
• 关键词: Address; Adenosine; Amino Acids; Binding; Biochemical; Biological; Cell Death; Cell Death Signaling Process; Cell Nucleus; Cell Survival; Chromatin; Code; Complex; Data; Disease; Embryo; Embryo Deaths; Embryonic Development; Enzymes; Equilibrium; Functional RNA; Gene Expression; Genetic Transcription; Goals; Health; Human; Infectious Arthritis; Inflammation; Inosine; Knockout Mice; Lead; Malignant - descriptor; Mammalian Cell; Mediating; MicroRNAs; Mus; Mutate; NF-kappa B; NFKB Signaling Pathway; Pathologic; Pathologic Processes; Pathology; Pathway interactions; Peptides; Phenotype; Physiological; Protein Binding Domain; Protein Fragment; Proteins; RNA; RNA Degradation; RNA Editing; Regulation; Reporting; Research; Role; Science; Signal Pathway; Signal Transduction; Staging; TNF gene; Tertiary Protein Structure; Testing; Tissues; Transportation; Work; base; cytokine; dsRNA adenosine deaminase; expectation; experience; improved; in vivo; innovation; knockout gene; p65; protein complex; protein function; protein protein interaction; public health relevance

DESCRIPTION (provided by applicant): ADAR1 (Adenosine Deaminase Acting on RNA -1) is an essential protein required for cell survival and embryonic development. Knockout of the gene encoding for this protein in mice gives rise to rapid and wide-spread cell death resulting in embryo death. The critical role of ADAR1 in cell survival indicates a potent role for ADAR1 in regulation of the balance between cell survival and cell death under physiologic conditions, and suggests that dysregulation of ADAR1 could lead to pathology. The long-term goal of the research of ADAR1 is to define its function and mechanism in order to therapeutically target ADAR1 in pathologic processes. Our preliminary data has shown that ADAR1 directly interacts with the NF-kB pathway and up-regulates NF-kB activities through IKK protein complex in mammalian cells. The objective in this application is to identify the mechanism of ADAR1 in the regulation of NF-kB signaling. The central hypothesis is that ADAR1 modulates NF-kB mediated gene expression through a protein-protein interaction that contributes to regulation of the balance between cell death and cell survival. Two specific aims will be addressed. Aim #1: Identify the mechanism in NF-kB pathway by which ADAR1 regulates NF-kB target gene expression. The dynamic interaction of ADAR1 and IKK2 subunit upon cytokine stimulation will be defined. The biochemical and signaling consequence of the protein- protein interaction will be determined. Aim #2: Determine the functional domain in ADAR1 that binds to IKK2 and regulates NF-kB activities. The protein binding domain of ADAR1 will be determined through expressions of mutated and truncated ADAR1 in mammalian cells and detect their capabilities to bind IKK2 and regulate NF-kB target gene transcriptions. The positively or negatively functioning protein fragments will also be tested whether they rescue the cell death phenotype of ADAR1 deficiency. At the completion of these studies, it is expected that a new function of ADAR1 in NF-kB pathway and the related mechanism will be revealed. A functional domain in ADAR1 interacts with NF-kB signal pathway will be identified. This protein domain may lead to a discovery of a peptide which will be used to treat diseases due to unbalance NF-kB signaling and cell death. This study is expected to have an important impact in the fields of RNA editing, signal transduction, and diseases that involve cell death mechanisms and therefore contribute to improve human health. PUBLIC HEALTH RELEVANCE: This research has relevance to diseases related to NFkB signaling pathway, such as inflammation, arthritis, infectious and malignant diseases.

描述（由申请人提供）：ADAR 1（腺苷脱氨酶作用于RNA-1）是细胞存活和胚胎发育所需的必需蛋白质。在小鼠中敲除编码这种蛋白质的基因会引起快速和广泛的细胞死亡，导致胚胎死亡。ADAR 1在细胞存活中的关键作用表明，在生理条件下，ADAR 1在调节细胞存活和细胞死亡之间的平衡中具有有效作用，并表明ADAR 1的失调可能导致病理学。ADAR 1的研究的长期目标是明确其功能和机制，以便在病理过程中治疗ADAR 1。我们的初步研究结果表明，ADAR 1直接与NF-κ B通路相互作用，并通过IKK蛋白复合物上调哺乳动物细胞中NF-κ B活性。本申请的目的是鉴定ADAR 1在NF-κ B信号传导调节中的机制。核心假设是ADAR 1通过蛋白质-蛋白质相互作用调节NF-κ B介导的基因表达，该蛋白质-蛋白质相互作用有助于调节细胞死亡和细胞存活之间的平衡。将讨论两个具体目标。目的#1：确定ADAR 1调节NF-κ B靶基因表达的NF-κ B通路机制。将定义细胞因子刺激后ADAR 1和IKK 2亚基的动态相互作用。将确定蛋白质-蛋白质相互作用的生化和信号传导结果。目的#2：确定ADAR 1中与IKK 2结合并调节NF-κ B活性的功能结构域。ADAR 1的蛋白结合结构域将通过突变和截短的ADAR 1在哺乳动物细胞中的表达来确定，并检测其结合IKK 2和调节NF-κ B靶基因转录的能力。还将测试正功能或负功能蛋白片段是否挽救ADAR 1缺陷的细胞死亡表型。这些研究的完成有望揭示ADAR 1在NF-κ B通路中的新功能及其相关机制。ADAR 1与NF-κ B信号通路相互作用的功能域将被鉴定。该蛋白质结构域可能会导致一种肽的发现，该肽将用于治疗因NF-kB信号传导不平衡和细胞死亡而引起的疾病。这项研究有望在RNA编辑、信号转导和涉及细胞死亡机制的疾病领域产生重要影响，从而有助于改善人类健康。公共卫生关系：本研究对与NFkB信号通路相关的疾病，如炎症、关节炎、感染性疾病和恶性疾病具有相关性。