MECHANISMS OF ONCORNAVIRUS-INDUCED TRANSFORMATION

冠状病毒诱导的转化机制

• 批准号: 3165340
• 负责人: Larry Ray Rohrschneider
• 金额: $ 23.84万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-02-01 至 1994-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3165340
• 关键词: Retroviridae; Retroviridae disease; athymic mouse; autoradiography; cell adhesion; cell growth regulation; cell transformation; cytoskeletal proteins; electroporation; fibronectins; gel electrophoresis; gene expression; immunochemistry; immunofluorescence technique; molecular cloning; monoclonal antibody; oncogenes; phosphorylation; point mutation; protein tyrosine kinase; protooncogene; radiotracer; receptor; temperature sensitive mutant; tissue /cell culture; transfection; vinculin; viral carcinogenesis; virus genetics; western blottings

The long term objectives of this project are to understand the molecular mechanisms of transformation induced by the v-src and v- fms oncogenes. In studies on v-src, experiments will concentrate on studying the interaction of pp60src with cellular adhesion plaques and defining the potential role of these structures in normal and tumor cell growth. To look for a role of adhesion plaques in regulating cell growth, the necessity of adhesion plaque formation will be examined in competent induction experiments and the potential modification and involvement of the fibronectin receptor in growth induction will be explored. Within v-src transformed cells, the role and significance of the tyrosine phosphorylation of the fibronectin receptor beta subunit will be tested by both biochemical and molecular genetic techniques. Alteration in binding talin and fibronectin will be measured in an in vitro binding assay, and deletion and point mutations will be produced in the cloned beta subunit cDNA of the fibronectin receptor. Over expression of these mutants in normal or transformed cells will test for altered function. In addition the role of the fibronectin receptor (especially the alpha subunits) in determining the target tissue for metastasis will be explored via the cloning and expression of various alpha and beta subunits of the fibronectin receptor. Studies on fms will focus on mechanisms of activation of the murine c-fms proto-oncogene. Mutations, over expression, and expression in inappropriate cell type will be examined both in vitro and in vivo. Also, within the v-fms oncogene, molecular biology techniques will be used to identify signals for endocytosis and ask whether this step is necessary for transformation. And finally, the function of the 70 amino acid insert in the tyrosine kinase domain of v-fms will be probed by deletion and substitution analysis. Overall these studies will enhance our understanding of the mechanism of neoplastic transformation of both solid and hematopoietic tumors.

该项目的长期目标是了解 V-SRC和V-诱导的转化的分子机制 FMS癌基因。 在V-SRC的研究中，实验将集中 研究PP60SRC与细胞粘附的相互作用 斑块并定义这些结构在 正常和肿瘤细胞生长。 寻找粘附的作用 调节细胞生长的斑块，粘附斑块的必要性 将在合理的归纳实验中检查形成，并 纤连蛋白的潜在修饰和参与 将探索生长诱导中的受体。 在V-SRC中 转化的细胞，酪氨酸的作用和意义 纤连蛋白受体β亚基的磷酸化将是 通过生化和分子遗传技术测试。 结合塔林和纤连蛋白的改变将在 体外结合测定，缺失和点突变将是 在纤连蛋白的克隆β亚基cDNA中产生 受体。 这些突变体在正常或 转化的单元将测试功能改变的功能。 另外 纤连蛋白受体的作用（尤其是α亚基） 在确定转移的目标组织时 通过各种α和β亚基的克隆和表达 纤连蛋白受体的。 FMS的研究将重点放在鼠的激活机制上 C-FMS原始癌基因。 突变，表达和表达 在不适当的细胞类型中，将在体外和 体内。 同样，在VM癌基因中，分子生物学 技术将用于识别内吞作用的信号并询问 是否需要此步骤进行转换。 最后， 70氨基酸插入酪氨酸激酶的功能 V-FM的域将通过删除和替换来探测 分析。 总的来说，这些研究将增强我们对 固体和固体肿瘤转化的机制 造血肿瘤。