SIGNAL TRANSDUCTION AND GENE INDUCTION IN T LYMPHOCYTES

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 钙调磷酸酶和几种组成型和诱导型激酶。NFAT蛋白发挥作用 在调节细胞诱导基因转录中的主要作用是 免疫系统在免疫反应过程中，也有牵连， 调节各种生理和病理生理过程的基因 在不同的非免疫细胞类型包括心脏、骨骼 肌肉、脂肪细胞和某些神经元细胞类型。长期目标是 项目的目的是了解 NFAT转录因子的激活和功能受到调节。其 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，探讨NFAT在原发性T细胞中的功能 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， 对照;开发靶向NFAT-钙调磷酸酶的细胞渗透试剂 互动，这可以用来检查NFAT在各种不同的作用， 生物过程;并与已建立的结构实验室合作 以获得NFAT 1-钙调磷酸酶复合物的X射线晶体结构。在Aim中 2，我们将确定调节NFAT家族的组成性激酶， 成员NFAT 1，并确定它们是否也磷酸化一个或多个 其他钙调节的NFAT。我们的目标是问一个人 NFAT家族的成员由共同或不同的机制调节。在 目的3，我们将通过以下方法来确定B y NFAT 1转录的调控机制： 定义N-末端的诱导磷酸化的作用， 反式激活结构域，N-末端反式激活中的LDFS基序 结构域，以及增强NFAT 1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 靶向NFAT蛋白的策略可适用于许多类型的 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。

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.