SIGNAL TRANSDUCTION AND GENE INDUCTION IN T LYMPHOCYTES

T 淋巴细胞中的信号转导和基因诱导

• 批准号: 6717685
• 负责人: Anjana Rao
• 金额: $ 47.3万
• 依托单位: IMMUNE DISEASE INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6717685
• 关键词: T lymphocyte; X ray crystallography; biological signal transduction; calcineurin; gene induction /repression; genetically modified animals; laboratory mouse; phosphorylation; protein kinase; protein structure; tissue /cell culture; transcription factor

DESCRIPTION (provided by applicant): Transcription factors of the NFAT family are highly-phosphorylated proteins whose functions are controlled by a balance between the calcium/ calmodulin-regulated serine/ threonine phosphatase calcineurin and several constitutive and inducible kinases. NFAT proteins play a major role in the regulating the transcription of inducible genes by cells of the immune system during an immune response, and have also been implicated in regulating various physiological and pathophysiological processes gene transcription in diverse non-immune cell types including heart, skeletal muscle, adipocytes, and certain neuronal cell types. The long-term goal of this project is to arrive at an understanding of the mechanisms by which the activation and function of NFAT transcription factors are regulated. Its specific aims are as follows. In Aim 1, we will develop reagents targeting the NFAT-calcineurin interaction, with which to explore NFAT functions in primary T cells. We will analyze transgenic mice in which a selective peptide inhibitor of the NFAT-calcineurin interaction, GFP-VIVIT, is expressed under tetracycline control; develop cell-permeant reagents that target the NFAT-calcineurin interaction, which can later be used to examine the role of NFAT in various biological processes; and collaborate with an established structural laboratory to obtain the X-ray crystal structure of the NFAT1-calcineurin complex. In Aim 2, we will identify the constitutive kinases that regulate the NFAT family member NFAT1, and determine whether they also phosphorylate one or more of the other calcium-regulated NFATs. Our objective is to ask whether the individual members of the NFAT family are regulated by common or distinct mechanisms. In Aim 3, we will define the mechanisms regulating transcription b y NFAT1, by defining the roles of inducible phosphorylation of the N-terminal transactivation domain, the LDFS motif in the N-terminal transactivation domain, and the modification that potentiates NFAT1 association with the histone acetyltransferases p300 and CBP. These experiments will increase our understanding of the molecular mechanisms of NFAT regulation. Therapeutic strategies that target NFAT proteins may have applicability in many types of pathological situations, including cardiac hypertrophy, transplant rejection, asthma and allergy, and autoimmune disease.

描述（由申请人提供）：NFAT 家族的转录因子 是高度磷酸化的蛋白质，其功能由平衡控制 钙/钙调蛋白调节的丝氨酸/苏氨酸磷酸酶之间 钙调神经磷酸酶和几种组成型和诱导型激酶。 NFAT 蛋白质发挥作用 在调节细胞诱导基因转录中发挥重要作用 免疫反应期间的免疫系统，并且也与 调节多种生理和病理生理过程的基因 在多种非免疫细胞类型（包括心脏、骨骼细胞）中转录 肌肉、脂肪细胞和某些神经元细胞类型。本次活动的长远目标 项目的目的是要了解 NFAT 转录因子的激活和功能受到调节。它是 具体目标如下。在目标 1 中，我们将开发针对 NFAT-钙调神经磷酸酶相互作用，用于探索原发性 T 中的 NFAT 功能 细胞。我们将分析转基因小鼠，其中选择性肽抑制剂 NFAT-钙调磷酸酶相互作用的 GFP-VIVIT 在四环素下表达 控制;开发针对 NFAT-钙调神经磷酸酶的细胞渗透试剂 相互作用，稍后可用于检查 NFAT 在各种中的作用 生物过程；并与成熟的结构实验室合作 获得 NFAT1-钙调神经磷酸酶复合物的 X 射线晶体结构。瞄准 2、我们将鉴定调节NFAT家族的组成型激酶 NFAT1 成员，并确定它们是否也磷酸化一个或多个 其他钙调节 NFAT。我们的目标是询问个人是否 NFAT 家族的成员受到共同或不同机制的调节。在 目标 3，我们将定义 NFAT1 调节转录的机制，通过 定义 N 末端诱导磷酸化的作用 反式激活结构域，N 端反式激活中的 LDFS 基序 域，以及增强 NFAT1 与 组蛋白乙酰转移酶 p300 和 CBP。这些实验将增加我们的 了解 NFAT 调节的分子机制。治疗性 靶向 NFAT 蛋白的策略可能适用于多种类型的疾病 病理情况，包括心脏肥大、移植排斥、 哮喘和过敏，以及自身免疫性疾病。