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。

基本信息

批准号：
8875012
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
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-01-15 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8875012
关键词：
1-Phosphatidylinositol 3-Kinase Affect Anaphylaxis Applications Grants Asthma Autoimmune Diseases B-Cell Activation 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 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

项目摘要

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-C2ß) 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-C2ß 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-C2ß, 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-C2ß. To determine the mechanism(s) whereby TRIM27 regulates PI3K-C2ß and how this is affected by TCR activation, we will determine in SA1, (Ai) the type of TRIM27 mediated ubiquitination of PI3K-C2ß; (Aii) the lysine residues on TRIM27 and PI3K-C2ß that are ubiquitinated; and (Aiii) the regions or domains on TRIM27 and PI3K-C2ß that mediate their association. In 1B, we will determine: (Bi) if TRIM27 regulates TCR stimulated PI3K-C2ß's kinase activity and/or degradation, or whether (Bii) association of TRIM27 with PI3K-C2ß 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-C2ß 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-C2ß is modulated by TCR stimulation. In (C), the role of TRIM27 stimulated SUMOylation of PI3K-C2ß 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-C2ß 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.

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-C2ß）导致PI3P产生，这是PI3P产生的，这是随后激活哺乳动物酶激酶，然后激活了Kinaase 3. kina kinoscase 3.我们确定了含有蛋白27（TRIM27）的三方基序是PI3K-C2ß和KCA3.1的新的负调节剂。 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-C2ß的机制，其物理意义，并随后作为KCA3.1在淋巴细胞和体内的淋巴细胞和肥大细胞激活中的负调节剂。我们有证据表明TRIM27功能E3连接酶和泛素化并抑制PI3K-C2ß。为了确定TRIM27调节PI3K-C2ß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-C2ß的泛素化的类型；（AII）TRIM27和PI3K-C2ß上的赖氨酸残基；（AIII）TRIM27和PI3K-C2ß上介导其关联的区域或域。在1B中，我们将确定：（bi）如果TRIM27调节TCR刺激PI3K-C2ß的激酶活性和/或降解，或者（BII）TRIM27与PI3K-C2ß的（BII）是否受TCR刺激的影响；（BIII）是否将TRIM27募集到免疫突触（IS），其在IS形成和/或募集PI3K-C2ß中的作用是在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-C2ß的泛素化是通过TCR模拟调节的。在（c）中，将评估TRIM27刺激的Sumoylation的作用。我们已经产生了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。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-C2ß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。