TRIM27 is a new negative regulator of CD4 T cells and Mast cells.

TRIM27 是 CD4 T 细胞和肥大细胞的新型负调节因子。

• 批准号: 8218480
• 负责人: EDWARD Y SKOLNIK
• 金额: $ 32.96万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-15 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8218480
• 关键词: 1-Phosphatidylinositol 3-Kinase; Affect; Anaphylaxis; Applications Grants; Asthma; Autoimmune Diseases; B-Cell Activation; Binding; Boxing; CD4 Positive T Lymphocytes; Calcium-Activated Potassium Channel; Cells; Coiled-Coil Domain; Cutaneous; Development; Failure; Feedback; Fingers; Generations; Goals; Graft Rejection; Helper-Inducer T-Lymphocyte; Histidine; Human; Hypersensitivity; Immune system; In Vitro; Inflammation; Lead; Lymphocyte; Lysine; Mediating; Mus; Names; Nucleoside diphosphate kinase B; Pathway interactions; Phosphoric Monoester Hydrolases; Phosphotransferases; Physiological; Potassium Channel; Protein Family; Proteins; Receptor Activation; Recruitment Activity; Regulation; Regulatory T-Lymphocyte; Role; Signal Pathway; Signaling Molecule; Structure of thyroid parafollicular cell; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; TRIM Motif; Th1 Cells; Ubiquitination; Zinc; allergic response; follow-up; immunological synapse; immunological synapse formation; in vivo; inhibitor/antagonist; insight; mast cell; member; myotubularin; phosphatidylinositol 3-phosphate; phosphohistidine; prevent; protein-histidine kinase; response; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): The K+ channel KCa3.1 is required for Ca2+ influx and the subsequent activation of B, T, and mast cells. Our studies have found that T cell receptor (TCR) activation leads to activation of KCa3.1 by activating the class 2 phosphatidylinositol 3 kinase C2 Beta (PI3K-C2¿) leading to the generation of PI3P, which is required for the subsequent activation of a mammalian histidine kinase, Nucleoside Diphosphate Kinase B, that then activates KCa3.1. We identified the Tripartate Motif containing protein 27 (TRIM27) as a new negative regulator of PI3K-C2¿ and KCa3.1. TRIM27 is a member of a large family of proteins characterized by the presence of a tripartite motif, consisting of a RING finger, B box, and coiled coil (CC) domains. In this proposal, we will determine the mechanisms whereby TRIM27 regulates PI3K-C2¿, its physiological significance, and subsequent role as a negative regulator of KCa3.1 in lymphocyte and mast cell activation in vitro and in vivo. We have evidence that TRIM27 functions an E3 ligase and ubiquitinates and inhibits PI3K-C2¿. To determine the mechanism(s) whereby TRIM27 regulates PI3K-C2¿ and how this is affected by TCR activation, we will determine in SA1, (Ai) the type of TRIM27 mediated ubiquitination of PI3K-C2¿; (Aii) the lysine residues on TRIM27 and PI3K-C2¿ that are ubiquitinated; and (Aiii) the regions or domains on TRIM27 and PI3K-C2¿ that mediate their association. In 1B, we will determine: (Bi) if TRIM27 regulates TCR stimulated PI3K-C2¿ 's kinase activity and/or degradation, or whether (Bii) association of TRIM27 with PI3K-C2¿ is affected by TCR stimulation; (Biii) whether TRIM27 is recruited to the immunological synapse (IS), its role in IS formation and/or the recruitment PI3K-C2¿ to IS following TCR activation; (Biv) whether TRIM27 modulates the PI3P levels in activated T cells, or (Bv) whether TRIM27 autoubiquitination, or ubiquitination of PI3K-C2¿ is modulated by TCR stimulation. In (C), the role of TRIM27 stimulated SUMOylation of PI3K-C2¿ will be assessed. We have generated TRIM27-/- mice and have evidence that KCa3.1 channel activity and TCR- stimulated Ca2+ flux are increased in TRIM27-/- Th1 cells. In SA2 (A), we will extend these studies to other CD4 helper T cell subsets, and determine whether Th2, Th17, and Treg differentiation and/or function are altered in cells isolated from TRIM27-/- mice. In (B), we will determine the downstream signaling pathways regulated by TRIM27, and (C) whether TRIM27-/- mice are predisposed to autoimmune disease. In (D) we will undertake a nonbiased approach to identify other targets of TRIM27 ubiquitination and regulation We found that TRIM27 functions to negatively regulate FceR1 stimulated KCa3.1 channel activity and Ca2+ flux in mast cells. We will determine in SA3: (A) if PI3K-C2¿ mediates FceR1 stimulated activation of KCa3.1 and whether this is modulated by TRIM27; (B) whether effectors functions of TRIM27-/- mast cells are increased; (3C) the signaling pathways regulated by TRIM27 in mast cells, and/or (3D) whether TRIM27-/- mice have an increased anaphylactic response to both passive cutaneous and systemic anaphylaxis. PUBLIC HEALTH RELEVANCE: Negative regulators of the adaptive and innate immune system, as well as mast cells, are critical to limit inflammation and to prevent allergic responses and autoimmune disease. Failure to down regulate these responses appropriately can lead to autoimmune disease and allergy, underscoring the critical role in understanding the signaling molecules that negatively regulate these cells. We have identified TRIM27 as a new negative regulator of CD4 T and mast cells. Understanding TRIM27's role in turning off these cells is likely to shed important insights into underlying mechanisms related to autoimmune disease and allergy.

描述（由适用提供）：Ca2+影响和随后的B，T和MAST细胞的激活需要K+通道KCA3.1。我们的研究发现，T细胞受体（TCR）激活通过激活2类磷脂酰肌醇3激酶C2β（PI3K-C2）导致PI3P的产生，这是哺乳动物酶激酶，核糖苷的随后激活Kinaper Kinapase 3. kinapase3. kinapase3.我们确定了含有蛋白27（TRIM27）的三方基序是PI3K-C2的新的负调节剂和KCA3.1。 TRIM27是大型蛋白质家族的成员，其特征是存在三方基序，由环手指，b盒和盘绕线圈（CC）域组成。在此提案中，我们将确定TRIM27调节PI3K-C2的机制，其物理意义以及随后作为KCA3.1在淋巴细胞和体内激活淋巴细胞和肥大细胞激活中的负调节剂的作用。我们有证据表明TRIM27功能E3连接酶并泛素化并抑制PI3K-C2。为了确定TRIM27调节PI3K-C2¿的机制，以及如何受TCR激活影响，我们将在SA1中确定（AI）TRIM27 TRIM27介导的PI3K-C2的泛素化的类型； （AII）泛素化的TRIM27和PI3K-C2“赖氨酸残基； （AIII）TRIM27和PI3K-C2上的区域或域，可以介导其关联。在1B中，我们将确定：（bi）如果TRIM27调节TCR刺激PI3K-C2？的激酶活性和/或降解，或者（BII）TRIM27与PI3K-C2的（BII）是否受TCR刺激的影响； （biii）是否将TRIM27招募到免疫突触（IS），其在IS形成和/或募集PI3K-C2上的作用是在TCR激活之后； （BIV）TRIM27是在活化的T细胞中调节PI3P水平，还是（bv）TRIM27自相传制或PI3K-C2的泛素化是通过TCR模拟调节的。在（c）中，TRIM27刺激PI3K-C2的Sumoylation的作用我们产生了TRIM27 - / - 小鼠，并有证据表明KCA3.1通道活性和TCR刺激的Ca2+通量在TRIM27 - / - TH1细胞中增加了。在SA2（a）中，我们将将这些研究扩展到其他CD4辅助T细胞子集，并确定在从TRIM27 - / - 小鼠中分离出的细胞中TH2，TH17和TREG分化和/或功能是否改变。在（b）中，我们将确定由TRIM27调节的下游信号通路，以及（c）Trim27 - / - 小鼠是否倾向于自身免疫性疾病。在（d）中，我们将采用一种无偏的方法来识别TIRM27泛素化和调控的其他靶标，我们发现TRIM27在肥大细胞中刺激FCER1刺激FCER1刺激的KCA3.1通道活性和Ca2+通量。我们将在SA3中确定：（a）如果PI3K-C2介导FCER1刺激了KCA3.1的激活，以及这是否由TRIM27调节； （b）TRIM27 - / - 肥大细胞的生效功能是否增加； （3C）在肥大细胞中由TRIM27调节的信号通路和/或（3D）是否对被动皮肤和全身性过敏反应的过敏反应增加。 公共卫生相关性：适应性和先天免疫系统的负调节剂以及肥大细胞，对于限制注射和防止过敏反应和自身免疫性疾病至关重要。不适当地调节这些反应可能会导致自身免疫性疾病和过敏，从而强调在理解负调节这些细胞的信号分子中的关键作用。我们已经将TRIM27确定为CD4 T和肥大细胞的新负调节剂。了解Trim27在关闭这些细胞中的作用可能会将重要的见解转化为与自身免疫性疾病和过敏有关的潜在机制。