THE FMS ONCOGENE

FMS癌基因

• 批准号: 3176249
• 负责人: CHARLES J SHERR
• 金额: $ 18.64万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1989-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3176249
• 关键词: complementary DNA; feline leukemia /sarcoma virus; gene expression; genetic library; genetic manipulation; genetic recombination; genetic transcription; genetic translation; human tissue; molecular cloning; nucleic acid sequence; oncogenes; oncogenic virus; point mutation; tissue /cell culture; viral leukemogenesis; virus RNA

The retroviral oncogene, v-fms, was acquired by genetic recombination between a filine leukemia virus (FeLV) and proto-oncogene sequences (c-fms) of normal cat cells. The v-fms gene is found in only one strain of feline sarcoma virus (FeSV) and encodes a transforming glycoprotein of unknown function. The product of the c-fms gene has not been identified, nor has its relationship to the viral transforming protein been elucidated. This proposal describes the detailed characterization of the v-fms-coded glycoprotein, defines the first steps in identifying the product of the cellular proto-oncogene, and attempts to clarify the role of these proteins in malignant transformation. Using the recently determined nucleotide sequence of a biologically active clone of FeSV DNA, we propose to place site-directed mutations in the v-fms gene. In particular, mutations will be directed to (i) limited regions of the v-fms gene which show unexpected homology to v-onc genes of the tyrosine kinase gene family, and to (ii) signal sequences which are presumed to play a role in the subcellular localization of the transforming glycoprotein. The transforming activity of altered genes will be assayed by DNA transfection onto cultured cells. Cotransfection with dominant selectable markers will be used to isolate nontransforming variants whose encoded products will be biochemically compared to those of the wild type transforming gene. The induced biochemical alterations in the v-fms gene product will be correlated with the topology of the protein and its transforming activity. In parallel studies, genomic and cDNA clones of the c-fms gene will be used to predict the primary structure of its product, thereby facilitating its identification and characterization. Recombinant DNA molecules between c-fms and v-fms will be prepared in an effort to activate the latent transforming potential of the cellular proto-oncogene. It is anticipated that a detailed characterization of domains in the v-fms-coded glycoprotein important in post-translational processing, intracellular transport, and transformation will provide the basis for activating the cellular proto-oncogene and determining its function. The latter studies will emphasize the structure and expression of the human c-fms proto-oncogene and its potential role in cancer.

逆转录病毒癌基因v-fms是通过基因重组获得的。 丝状白血病病毒(FeLV)和原癌基因序列(c-FMS)之间的关系 正常的猫细胞。V-fms基因只在一种猫科动物身上发现。 肉瘤病毒(FeSV)，编码一种未知的转化糖蛋白 功能。C-fms基因的产物还没有被鉴定，也没有。 它与病毒转化蛋白的关系已被阐明。 该方案描述了v-FMS编码的详细特征 糖蛋白，定义了鉴定产物的第一步 细胞原癌基因，并试图阐明这些蛋白的作用 在恶变过程中。使用最近确定的核苷酸 一个FeSV DNA的生物活性克隆的序列，我们建议放置 V-fms基因的定点突变。特别是，突变将 被定向到(I)v-fms基因的有限区域，这些区域显示出意想不到的 与酪氨酸激酶基因家族的v-onc基因同源，并与(Ii) 被认为在亚细胞中起作用的信号序列 转化糖蛋白的定位。转型活跃度 将通过将DNA导入培养细胞来检测改变基因的数量。 与显性可选择标记的共转染将被用于分离 其编码产物将是生化的非转化变异体 与野生型转化基因进行比较。被诱导的 V-fms基因产物的生化变化将与 蛋白质的拓扑结构及其转化活性。并行的 研究，c-fms基因的基因组和cdna克隆将用于预测 其产品的一级结构，从而促进其 鉴定和表征。重组DNA分子在 将制备C-FMS和V-FMS，以努力激活潜伏期 细胞原癌基因的转化潜能。这是意料之中的 V-FMS编码的糖蛋白中结构域的详细特征 在翻译后处理、细胞内运输和 转化将为激活细胞提供基础 原癌基因及其功能的测定。后一项研究将 强调人c-fms原癌基因的结构和表达 以及它在癌症中的潜在作用。