The Role of PIAS1 in Immune Regulation

描述(申请人提供)：STAT1和核因子-kappaB在免疫系统的调节中发挥重要作用。STAT和NF-kappaB活性的异常调节与多种人类免疫疾病有关。STAT1和核因子-kappaB的激活在启动先天免疫反应中起着核心作用。我实验室的研究已经确定PIAS1(激活的STAT1的蛋白抑制物)是STAT1和NF-kappaB的常见负调节因子。最近，我们培育出了PIAS1缺失小鼠。我们的初步研究已经确定了PIAS1在免疫反应调节中的生理作用。这项研究计划的总体目标是研究PIAS1在免疫调节中的作用。这些研究将增强我们设计治疗传染病的新治疗策略的能力。 我们将描述PIASL基因敲除小鼠的免疫学表型，并研究PIAS1在病原体天然免疫中的作用。我们将研究PIAS1的信号特异性。基因芯片分析将被用来确定PIAS1在调节干扰素诱导的STAT1依赖的基因激活中的特异性。我们将使用体外和体内DNA结合试验来检测STAT1结合亲和力对PIAS1特异性的潜在影响。此外，我们将通过基因打靶和RNA干扰方法测试PIASy是否影响PIAS1在STAT1依赖的基因激活中的特异性。我们将探讨PIAS1介导的免疫调节作用的分子机制。具体地说，我们将使用基因打靶策略来研究PIAS1相扑连接酶活性在PIAS1介导的基因调控中的生理意义。PIAS1介导的NF-kappaB调节的意义和PIAS1调节的STAT1依赖的基因激活在抗病毒反应中的参与将被检测。

DESCRIPTION (provided by applicant): Stat1 and NF-kappaB play important roles in the regulation of the immune system. Abnormal regulation of STAT and NF-kappaB activities has been associated with various human immune disorders. The activation of Stat1 and NF-kappaB has a central role in initiating the innate immune response. Studies from my laboratory have identified PIAS1 (protein inhibitor of activated Stat1) as a common negative regulator of Stat1 and NF-kappaB. Recently, we have generated PIAS1 null mice. Our preliminary studies have identified a physiological role of PIAS1 in the regulation of immune responses. The overall goal of this research proposal is to study the role of PIAS1 in immune regulation. These studies will enhance our ability to design novel therapeutic strategies for the treatment of infectious diseases. We will characterize the immunological phenotypes of Piasl knockout mice and examine the role of PIAS1 in innate immunity to pathogens. We will study the signaling specificity of PIAS1. Microarray analysis will be performed to define the specificity of PIAS1 in regulating the IFN-induced Stat1-dependent gene activation. We will examine the potential effect of Stat1 binding affinity on the specificity of PIAS1 using in vitro and in vivo DNA binding assays. In addition, we will test if PIASy affects the specificity of PIAS1 in Stat1-dependent gene activation by gene targeting and RNA interference approaches. We will examine the molecular mechanisms involved in the PIASl-mediated effect in immune regulation. Specifically, we will investigate the physiological importance of the PIAS1 SUMO ligase activity in PIASl-mediated gene regulation using a gene targeting strategy. The significance of the PIASl-mediated NF-kappaB regulation and the involvement of PIASl-regulated Stat1-dependent gene activation in antiviral response will be examined.