RAF-1 KINASE AND CYTOKINE MEDIATED SIGNALING

对小分子量分子如何传递 从细胞外到细胞核的信息 这对我们理解癌症、正常细胞功能以及 发展 这项研究的目的是确定 细胞因子刺激Raf-1激酶，从而刺激 这是一条可以导致细胞核转录改变的途径。 在 为了理解Raf-1激活的机制， 研究：1）Raf-1激酶与JAK家族的相互作用， 激酶（已知由细胞因子激活）2）Raf-1的变化 磷酸化引起的JAK激酶和3）的影响， 磷酸化对Raf-1激酶活性的影响。 解决这些 问题Raf-1（或各种修改后的形式）将表示为 使用杆状病毒在昆虫细胞中检测JAK家族的不同成员 系统 使用该系统，将有可能定义哪些域 是蛋白质相互作用所必需的， 磷酸化反应的结果，以及磷酸化反应有什么影响 Raf-1的活动。 在昆虫细胞中学到的信息将 在用细胞因子刺激的CTLL细胞中证实， 使用杆状病毒系统获得的结果是生物学相关的。

The understanding of how small molecular weight molecules transmit information from the outside of cells to the nuclei of those cells is critical for our understanding of cancer, normal cell functioning, and development. The aim of this fellowship is to determine the molecular mechanisms by which cytokines stimulate Raf-1 kinase, thereby stimulating a pathway which can result in altered transcription in the nucleus. In order to understand the mechanism of Raf-1 activation the following will be examined: 1) the interaction of Raf-1 kinase with the JAK family of kinases (known to be activated by cytokines) 2) the change in Raf-1 phosphorylation caused by the JAK kinases and 3) the effect the phosphorylations have on the kinase activity of Raf-1. To address these questions Raf-1 (or various modified forms) will be expressed with different members of the JAK family in insect cells using the baculovirus system. Using this system it will be possible to define which domains are needed for the proteins to interact, what effects on phosphorylation result from the interaction, and what effects the phosphorylations have on Raf-1 activity. The information learned in the insect cells will be confirmed in CTLL cells stimulated with cytokines to insure that the results obtained using the baculovirus system are biologically relevant.