UBIQUITYLATION AND THE REGULATION OF IMMUNE HOMOSTASIS

普遍性和免疫稳态的调节

• 批准号: 8363762
• 负责人: AVERIL I MA
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363762
• 关键词: Biochemical; Cells; Data; Disease; Enzymes; Eukaryotic Cell; Funding; Genetic; Grant; Homeostasis; Immune; Immune response; Inflammatory Response; Intestines; Link; Mass Spectrum Analysis; Mediating; Modification; Mus; Myeloid Cells; National Center for Research Resources; Principal Investigator; Proteins; Receptor Signaling; Regulation; Research; Research Infrastructure; Resources; Signal Transduction; Signaling Protein; Source; TRAF6 gene; Testing; Toll-like receptors; United States National Institutes of Health; cost; in vivo; insight; microbial; novel; pathogen; prevent; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Toll like receptors (TLRs) on eukaryotic cells mediate the recognition of microbial molecules and regulate host responses to microbial pathogens. Proper regulation of TLR signals not only restricts the intensity and duration of inflammatory responses, but may also maintain immune homeostasis in the intestine, where a plethora of microbial pathogens co-exist with host cells. Recent studies from our labs indicate that A20 is a novel protein that is required for terminating TLR induced signals on myeloid cells and for preventing excessive inflammatory responses in vivo. Our preliminary data suggest that A20 is essential for restricting TLR signals in vivo. Moreover, our findings suggest that A20 may be a novel enzyme that performs this critical function by directly modulating the ubiquitylation status of TLR signaling proteins such as TRAF6. These modifications may cause both de-activation and degradation of TLR signaling proteins. These exciting preliminary findings provide us with unique opportunities to test our central hypothesis that A20 regulates ubiquitylation of signaling proteins, TLR signals, and immune homeostasis. This application represents a synergistic effort between the genetic and cellular expertises of the Ma lab and the biochemical expertise of the Pickart lab to determine: (i) whether A20 is essential for regulating TLR responses in mice; (ii) whether A20 regulates ubiquitylation of critical TLR signaling proteins; and (iii) how A20 recognizes ubiquitylated proteins and may function as both a de-ubiquitylating enzyme and an E3. In the latter approach, mass spectroscopic analysis will be essential for understanding how A20 modifies ubiquitylated proteins. Selected TLR signaling proteins will be examined as putative substrates, and both A20's de-ubiquitylating and ubiquitylating functions will be assessed on candidate substrates. Results from these studies promise to yield significant insights into how A20 links the regulation of ubiquitylation with disease.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 真核细胞上的 Toll 样受体 (TLR) 介导微生物分子的识别并调节宿主对微生物病原体的反应。 TLR 信号的正确调节不仅可以限制炎症反应的强度和持续时间，还可以维持肠道内的免疫稳态，肠道内有大量微生物病原体与宿主细胞共存。我们实验室的最新研究表明，A20 是一种新型蛋白质，是终止骨髓细胞上 TLR 诱导信号和防止体内过度炎症反应所必需的。我们的初步数据表明 A20 对于限制体内 TLR 信号至关重要。此外，我们的研究结果表明，A20 可能是一种新型酶，它通过直接调节 TLR 信号蛋白（如 TRAF6）的泛素化状态来执行这一关键功能。这些修饰可能导致 TLR 信号蛋白失活和降解。这些令人兴奋的初步发现为我们提供了独特的机会来检验我们的中心假设，即 A20 调节信号蛋白、TLR 信号和免疫稳态的泛素化。该应用代表了 Ma 实验室的遗传和细胞专业知识与 Pickart 实验室的生化专业知识之间的协同努力，以确定：(i) A20 是否对于调节小鼠 TLR 反应至关重要； (ii) A20 是否调节关键 TLR 信号蛋白的泛素化； (iii) A20 如何识别泛素化蛋白并可同时充当去泛素化酶和 E3。在后一种方法中，质谱分析对于了解 A20 如何修饰泛素化蛋白至关重要。选定的 TLR 信号蛋白将作为假定底物进行检查，并且将在候选底物上评估 A20 的去泛素化和泛素化功能。这些研究的结果有望对 A20 如何将泛素化调节与疾病联系起来提供重要见解。