FUNCTIONS OF ADENOVIRUS PROTEINS IN TRANSFORMATION

腺病毒蛋白的转化功能

• 批准号: 3168787
• 负责人: JAMES B LEWIS
• 金额: $ 7.93万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-04-01 至 1990-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3168787
• 关键词: Adenoviridae; cell free system; gene expression; genetic manipulation; genetic mapping; genetic regulation; genetic transcription; immunological substance; molecular oncology; nucleic acid sequence; oncogenes; oncogenic virus; radiotracer; viral carcinogenesis; virus DNA; virus genetics

Transcription units E1a and E1b of the human adenovirus encode all of the functions needed for the oncogenic transformation of primary rodent cells to cell lines that are immortalized and have altered morphology and growth properties, both in vitro and in vivo. Knowledge of hos these viral genes function to effect such changes should permit elucidation of the molecular mechanisms by which normal cells become neoplastic ans should suggest strategies for controlling the growth of tumor cells. Five or more proteins are encoded by E1a and E1b. The E1a and E1b gene products that are necessary for the different aspects of oncogenic transformation will be identified by transfection of cells with appropriately mutated recombinant DNA molecules. The roles of these E1a and E1b proteins, in transformation will be further studied using antibodies generated by immunization with synthetic peptides corresponding to portions of individual proteins. These antibodies will be used to measure the synthesis and accumulation of E1a and E1b proteins and to study the compartmentalization of these proteins in infected and transformed cells, and to characterize the associations of the E1a and E1b proteins with cellular structures. Particular attention will be paid to the interaction of E1a proteins with nuclear structure. Specific antibodies to E1a proteins will be used to assay for site-specific interaction with viral DNA. Mutation will be made in the E1a-289R protein and used to correlate its transcription-enhancing function with its role in transformation and its localization in the nuclear matrix. These mutations will also be used to search for differential interaction of the E1a-289R protein with different viral promoters, and for possible differential interaction with the cellular genes that are targets of E1a action. Any mutants that interact with different subsets of these target genes would be useful in determining which targets are critically important for oncogenic transformation.

人腺病毒的转录单元E1a和E1b编码所有 原代啮齿动物细胞的致癌转化所需的功能 到永生并改变形态和生长的细胞系 体外和体内的特性。 这些病毒基因的知识 实现这种变化的功能应允许阐明分子 正常细胞变成肿瘤的机制应提示 控制肿瘤细胞生长的策略。 五个或更多 蛋白质由E1A和E1B编码。 E1A和E1B基因产品， 对于致癌转化的不同方面是必需的 通过适当突变重组的细胞转染鉴定 DNA分子。 这些E1A和E1B蛋白的作用，在转化中 将使用由免疫产生的抗体进一步研究 对应于单个蛋白质的部分的合成肽。 这些 抗体将用于测量E1a的合成和积累 和E1b蛋白质，并研究这些蛋白质的分隔 感染和转化的细胞，并表征 具有细胞结构的E1A和E1B蛋白。 特别关注 为E1A蛋白与核结构的相互作用支付。 E1A蛋白的特异性抗体将用于测定特定于位点的抗体 与病毒DNA的相互作用。 突变将在E1A-289R蛋白中进行 并用于将其转录增强功能与其在 转化及其在核基质中的定位。 这些突变 还将用于搜索E1A-289R的差异相互作用 具有不同病毒启动子的蛋白质，并且可能差异 与E1a作用靶标的细胞基因相互作用。 任何 与这些靶基因不同亚集相互作用的突变体将是 在确定哪些目标对致癌性至关重要的情况下有用 转型。