ANALYSIS OF RAF-ONCOGENE RELATED NOVEL PROTEIN KINASE GENE

我们继续分析一种新的蛋白激酶基因(称为c- 否)编码一种与RAF癌基因具有广泛同源性的蛋白质。 已经产生了针对激活肽和重组蛋白的抗体 片段，并被用来通过以下方式鉴定激酶蛋白 免疫沉淀和免疫印迹。初步实验已经 在多种细胞的细胞核中发现一种70-80kD的蛋白质 类型。在体外，信使核糖核酸的转录和翻译被用于 用于功能分析和鉴定细胞蛋白的激酶蛋白 底物。由于其不同寻常的碱性氨基酸在 调节结构域，该激酶似乎与其他核仁有关 RNA结合蛋白，尤其是HIV、Rev和Tat调节蛋白。 目前的实验针对的是定义生理催化 激酶蛋白的功能及其与核酸的结合 潜力。

We have continued our analysis of a novel protein kinase gene (termed c- NO) encoding a protein with extensive homology to the raf oncogene. Antibodies have been generated to kinase peptides and recombinant protein fragments and have been used to identify the kinase protein by immunoprecipitation and western blotting. Preliminary experiments have identified a protein of 70-80 Kd in the nucleus of a variety of cell types. In vitro mRNA transcription and translation are being used to make kinase protein for functional assays and to identify cellular protein substrates. Because of its unusual basic amino acid repeats in the regulatory domain, this kinase appears to be related to other nucleolar RNA binding proteins especially the HIV REV and TAT regulatory proteins. Current experiments are directed at defining the physiological catalytic function of the kinase protein and at determining its nucleic acid binding potential.