DYNAMICS OF C-FOS PROTEIN INTERACTIONS

我们的实验室最近表明，诱导c-fos的表达， 基因是细胞对广泛的跨膜信号传导的反应 包括生长因子、佛波醇酯和神经递质的试剂。 我们 拟研究Fos蛋白的性质及其诱导。 具体而言，我们建议： 1.测定Fos蛋白合成的时程， 生长后fos与第二种细胞蛋白p39的结合 在3 T3细胞中的因子诱导。 2.测定fos蛋白与 第39页。 从化学计量的变化中，我们将推导出 fos-p39复合物形成。 3.去突变fos蛋白。 我们将确定结合位点， fos的p39和比较这与改变fos蛋白的突变位点 转化能力，核定位，翻译后修饰 和表达自动调节的潜力。 这将开发一个 Fos蛋白的功能拓扑学。 4.通过分离p39与fos的复合物来纯化p39蛋白， 目的制备研究p39表达和结构的试剂。 5.为了检测fos蛋白在细胞凋亡中的负调节作用， fos启动子 6.检测其他生长因子诱导的Fos早期反应基因 使用反义技术检测转录阻遏物活性。 7.为了鉴定与fos调控序列相互作用的蛋白质， 新的DNA标记转移测定。

Our laboratory has recently shown that induction of expression of the c-fos gene is a cellular response to a wide range of transmembrane signaling agents including growth factors, phorbol esters, and neurotransmitters. We proposed to study properties of the fos protein and its induction. Specifically we propose: 1. To determine the time course of synthesis of fos protein and the association of fos with a second cellular protein, p39 following growth factor induction in 3T3 cells. 2. To determine the molecular weight of the complex of fos protein with p39. From changes in this stoichiometry we will deduce the dynamics of fos-p39 complex formation. 3. To mutagenize the fos protein. We will determine the binding site on fos for p39 and compare this with sites of mutation which alter fos protein transforming ability, nuclear localization, post-translational modification and potential for autoregulation of expression. This will develop a functional topology for the fos protein. 4. To purify the p39 protein through isolation of its complex with fos with the objective of preparing reagents to study p39 expression and structure. 5. To assay the fos protein for a negative regulatory role acting at the fos promoter. 6. To assay other growth factor induced early response genes for fos transcription repressor activity using the antisense technique. 7. To identify proteins interacting with fos regulatory sequences using a novel DNA label transfer assay.