NORMAL AND ABNORMAL INTERFERON ALPHA RECEPTORS

干扰素、细胞因子和生长因子发挥着重要的作用 针对病毒感染的第一道防线，在治疗不同 癌症的类型与免疫和造血的调节 系统。它们的影响几乎涉及到有机体的每一个系统 强调它们的广泛影响和阐明它们的重要性 信令机制。细胞因子、生长因子和IFN具有相似的 涉及受体结合和激活的信号机制 Jak和Stat(信号)的酪氨酸激酶和转录因子 转导和转录激活子)家族，导致 与基因转录激活有关的不同 细胞因子反应。 然而，人们对细胞因子的作用机制知之甚少。 受体激活Jak激酶和Stat因子。此应用程序 试图描述细胞因子受体用来 激活Jak激活酶。我们将检验类似的假设 存在于细胞因子受体胞浆区域的结构域可能是 用于JAK激酶的相互作用和激活。然而， 这个胞质结构域可能在激活的过程中扮演不同的角色 JAK1和JAK2通过不同的受体。在这个项目的第一个具体目标中 提案的重点是JAK1结合结构域的特征 存在于激活该激酶的不同细胞因子受体中。我们 还将描述Jak Kinase中所需的结构域 与不同的细胞因子受体相互作用。第二 为测试互动/的不同模型提出了具体目标 细胞因子受体和JAK激酶之间的激活，应激 细胞因子受体与JAK1和JAK2的相互作用/激活。 为了实现这些目标，我们提出了体内和体外研究，如 编码猪瘟病毒突变的GST融合蛋白的生产 合适的结合位点，双杂交系统，生产重组 使用体外翻译系统的蛋白质，瞬时和稳定 重组蛋白在不同哺乳动物细胞中的表达。这个 通过这项提议获得的结果将不仅对 了解IFNpha的信号通路，也通过 不同的细胞因子受体。

Interferons (IFN), cytokines and growth factors play important roles as a first defense against viral infections, in the treatment of different types of cancers and in the regulation of the immune and hematopoietic system. Their effects involve almost every system in the organism underscoring their broad effects and the importance of elucidating their signaling mechanisms. Cytokine, growth factors and IFNs have similar signaling mechanism that involve receptor binding and activation of tyrosine kinases and transcription factors of the Jak and Stat (Signal Transducers and Activators of Transcription) families, that results in the transcriptional activation of genes responsible for the different cytokine responses. However, little is known about the mechanisms by which cytokine receptors activate the Jak kinases and Stat factors. This application attempts to characterize the mechanisms used by cytokine receptors to activate the Jak kinases. We will test the hypothesis that similar domains present in the cytoplasmic region of cytokine receptors may be utilized for the interaction and activation of Jak kinases. However, this cytoplasmic domain may have different roles in the activation of Jak1 and Jak2 by distinct receptors. In the first Specific Aims of this proposal is focused on the characterization of the Jak1 binding domain present in different cytokine receptors that activate this kinase. We will also characterize the domains in the Jak kinases that are required for the interaction with different cytokine receptors. The second Specific Aim proposed to test different models for the interaction/ activation between cytokine receptors and Jak kinases, stressing the interaction/activation of cytokine receptors with Jak1 and Jak2. To achieve these goals we propose in vivo and in vitro studies such as the production of GST fusion protein encoding mutation of the appropriate binding sites, two hybrid system, production of recombinant proteins using in vitro translation systems, transient and stable expression of recombinant proteins in different mammalian cells. The results obtained with this proposal will be important not only to understand the signaling pathway of IFNalpha, but also signaling by different cytokine receptors.