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）：ADAR1（作用于RNA的腺苷脱氨酶-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导致快速且广泛的细胞死亡，从而导致胚胎死亡。 ADAR1 在细胞存活中的关键作用表明 ADAR1 在调节生理条件下细胞存活和细胞死亡之间的平衡方面发挥着重要作用，并表明 ADAR1 失调可能导致病理。 ADAR1研究的长期目标是明确其功能和机制，以便在病理过程中靶向ADAR1进行治疗。我们的初步数据表明，ADAR1 直接与 NF-kB 通路相互作用，并通过哺乳动物细胞中的 IKK 蛋白复合物上调 NF-kB 活性。本申请的目的是确定 ADAR1 在 NF-kB 信号传导调节中的机制。中心假设是 ADAR1 通过蛋白质-蛋白质相互作用调节 NF-kB 介导的基因表达，从而有助于调节细胞死亡和细胞存活之间的平衡。将解决两个具体目标。目标 #1：确定 ADAR1 调节 NF-kB 靶基因表达的 NF-kB 通路机制。 ADAR1 和 IKK2 亚基在细胞因子刺激下的动态相互作用将被定义。将确定蛋白质-蛋白质相互作用的生化和信号转导结果。目标 #2：确定 ADAR1 中与 IKK2 结合并调节 NF-kB 活性的功能域。 ADAR1的蛋白结合域将通过突变和截短的ADAR1在哺乳动物细胞中的表达来确定，并检测其结合IKK2和调节NF-kB靶基因转录的能力。阳性或阴性功能的蛋白质片段也将被测试是否可以挽救 ADAR1 缺陷的细胞死亡表型。这些研究完成后，有望揭示ADAR1在NF-kB通路中的新功能及相关机制。 ADAR1 中与 NF-kB 信号通路相互作用的功能域将被识别。该蛋白结构域可能会导致一种肽的发现，该肽将用于治疗由于 NF-kB 信号传导失衡和细胞死亡而导致的疾病。这项研究预计将在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。公共健康相关性：这项研究与 NFkB 信号通路相关的疾病相关，如炎症、关节炎、传染病和恶性疾病。