RAS P21 PROTEINS AND INDENTIFYING DBL ONCOGENE PRODUCT

RAS P21 蛋白和 DBL 癌基因产物的鉴定

• 批准号: 3963467
• 负责人: S K SRIVASTAVA
• 金额: --
• 依托单位: DIVISION OF CANCER EPIDEMIOLOGY AND GENETICS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3963467
• 关键词: conformation; gel electrophoresis; gene mutation; human tissue; lung neoplasms; molecular cloning; monoclonal antibody; mouse sarcoma virus; neoplasm /cancer genetics; oncogenes; viral carcinogenesis

Project 1: In our continued effort to understand the structure-function relationship of ras p21, we have made inframe deletions of 4-5 amino acids in the region of p21 protein as the epitope for anti-ras p21 monoclonal antibody, Y13-259. This antibody can revert the transforming effect of p21 and can also block the initiation of DNA synthesis. However, Y13-259 does not directly interfere with known biochemical properties of p21. These studies suggest that this antibody may affect some as yet undetected function of p21. To gain insights into the function of Y13-259 binding region of p21, these mutants are currently being analyzed for their biochemical and biological properties. Project 2: A novel oncogene, db1, isolated from a human diffuse B-cell lymphoma, has been previously shown to confer the transformed phenotype on NIH/3T3 cells. Our present studies were undertaken in an effort to identify the db1 oncogene product and to characterize some of its structural and functional properties. A 66,000 dalton protein (p66) has been identified in cells that have acquired db1 oncogene, while it is not present in cells transformed by other oncogenes or in untransformed NIH/3T3 cells. Furthermore, it is established that p66 is encoded by db1 oncogene. p66 is a phosphoprotein with serine a major site of phosphorylation and it is a cytoplasmic protein as judged by its subcellular distribution.

项目 1：我们不断努力理解结构-功能 ras p21 的关系，我们进行了 4-5 个氨基酸的框内删除 在 p21 蛋白区域作为抗 ras p21 单克隆抗体的表位 抗体，Y13-259。 该抗体可以恢复p21的转化作用 还可以阻断DNA合成的起始。 然而，Y13-259 确实 不直接干扰 p21 的已知生化特性。 这些 研究表明这种抗体可能会影响一些尚未被发现的 p21的功能。 深入了解 Y13-259 结合的功能 p21 区域，目前正在分析这些突变体的 生化和生物学特性。 项目 2：从人类弥漫性 B 细胞中分离出一种新的致癌基因 db1 淋巴瘤，之前已被证明会赋予转化的表型 NIH/3T3 细胞。 我们目前的研究是为了 鉴定 db1 癌基因产物并表征其一些 结构和功能特性。 66,000 道尔顿的蛋白质 (p66) 具有 在获得 db1 癌基因的细胞中被鉴定出来，而并非如此 存在于被其他癌基因转化的细胞或未转化的 NIH/3T3 中 细胞。 此外，确定p66是由db1编码的 癌基因。 p66 是一种磷蛋白，丝氨酸是其主要位点 磷酸化，根据其判断它是一种细胞质蛋白 亚细胞分布。