RAS GENES BY DOMINANT NEGATIVE MUTANTS

显性阴性突变体的 RAS 基因

• 批准号: 3460112
• 负责人: YU-WEN HWANG
• 金额: $ 10.38万
• 依托单位: INSTITUTE FOR BASIC RES IN DEV DISABIL
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-12-15 至 1995-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3460112
• 关键词: PC12 cells; autosomal dominant trait; biological products; biological signal transduction; cell differentiation; enzyme linked immunosorbent assay; gene induction /repression; gene interaction; gene mutation; genetic manipulation; genetic promoter element; guanine nucleotide binding protein; microinjections; microorganism genetics; molecular cloning; molecular genetics; neurotrophic factors; nucleic acid hybridization; oncogenes; radionuclides; tissue /cell culture; transfection

It has been demonstrated that activated ras genes play vital roles in many types of human tumor. The immediate ras gene family consists of three members, H-ras, K-ras and N-ras. The ras genes belong to a group of proteins collectively called guanine-nucleotide binding proteins. Guanine-nucleotide binding proteins require guanine-nucleotides for their proper functions and share several conserved features among them. Many guanine-nucleotide binding proteins are involved in transducing the stimuli signals to their targets; therefore, it is conceivable that the ras gene products have similar biological roles. At present, not much is known about the signal transduction pathway of ras genes. In addition to their apparent role in cell proliferation, ras genes also have been shown to induce terminal differentiation in some cell types, such as rat pheochromocytoma cell line PC12, a in vitro neuronal cell model. Recently, we have described a class of H-ras mutants which can dominantly suppress the action of the c-H-ras gene as well as several other oncogenes, such as src, fms, sis and fes. The basis for this type of trans-dominant suppression might be attributed to sequestering of the essential element required for c-H-ras activation. These mutants will provide an extremely valuable tool for investigating the signal transduction pathway of ras genes and the interactions of ras genes with other oncogenes. We propose to study the underlying mechanism of trans-dominant suppression and will also utilize the trans-dominant properties of these mutants to study the biological role of the ras genes. 1. We will determine the physical requirements of the H-ras p2l protein for trans-dominant suppression; 2. we will determine the effects of K-ras (or N-ras) mutants on c-H-ras and other oncogenes as mentioned previously; 3. we will utilize the trans-dominant mutants as the affinity probe for purifying the element that interacts with ras p2l protein and 4. we will investigate the role of the ras p2l protein in differentiating PC12 cells by trans-dominant mutants.

研究表明，激活的 ras 基因在许多疾病中发挥着重要作用。 人类肿瘤的类型。直接的ras基因家族由三个组成 成员：H-ras、K-ras 和 N-ras。 ras 基因属于一组 蛋白质统称为鸟嘌呤核苷酸结合蛋白。 鸟嘌呤核苷酸结合蛋白需要鸟嘌呤核苷酸才能发挥作用 它们具有适当的功能并共享一些保守的特征。许多 鸟嘌呤核苷酸结合蛋白参与刺激的转导 向目标发出信号；因此，可以想象，ras基因 产品具有相似的生物学作用。目前，人们对此知之甚少 ras基因的信号转导途径。除了他们明显的 ras 基因也被证明可以诱导细胞增殖 某些细胞类型的终末分化，例如大鼠嗜铬细胞瘤 细胞系PC12，体外神经元细胞模型。 最近，我们描述了一类 H-ras 突变体，它可以显性地 抑制 c-H-ras 基因以及其他几种癌基因的作用， 例如 src、fms、sis 和 fes。这种类型的反式显性的基础 抑制可能归因于基本元素的隔离 c-H-ras 激活所需。这些突变体将提供极其 研究 ras 信号转导途径的宝贵工具 基因以及ras基因与其他癌基因的相互作用。我们建议 研究跨显性抑制和意志的潜在机制 还利用这些突变体的反式显性特性来研究 ras 基因的生物学作用。 1. 我们将确定 H-ras p2l 蛋白的物理需求 反式显性抑制； 2.我们将确定K-ras（或 N-ras) c-H-ras 和前面提到的其他癌基因的突变体； 3. 我们将利用反式显性突变体作为亲和探针 纯化与 ras p2l 蛋白相互作用的元素，4. 我们将 研究ras p2l蛋白在分化PC12细胞中的作用 通过反式显性突变体。