MECHANISMS OF ONCOGENIC TRANSFORMATION BY ADENOVIRUSES

腺病毒致癌转化机制

• 批准号: 2088438
• 负责人: JAMES F WILLIAMS
• 金额: $ 18.71万
• 依托单位: CARNEGIE-MELLON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-08-01 至 1998-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2088438
• 关键词: Adenoviridae; MHC class I antigen; cell transformation; chimeric proteins; enzyme linked immunosorbent assay; flow cytometry; gene expression; host organism interaction; immunogenetics; immunoprecipitation; laboratory mouse; laboratory rat; mutant; natural killer cells; neoplastic transformation; oncogenes; oncogenic virus; oncoproteins; phenotype; protein structure function; tissue /cell culture; transcription factor; viral carcinogenesis; virus genetics; virus protein

The overall, long-term objective of this research program is to use genetic, immunological and molecular approaches to define the functions that the type 5 and type 12 adenovirus (Ad5 and Ad12) E1A and E1B proteins, and perhaps other viral proteins, play in transforming rat and mouse cells in vitro, and in regulating the events which occur in the early phase of the productive infection in human cells. A particular goal is to understand the basis for the strong oncogenicity of Ad12 in rodents and the striking difference in oncogenicity between that serotype and the non-oncogenic type 5 virus. To this end we have constructed and are characterizing Ad12 and Ad5-based viruses containing chimeric E1A genes, and have used some of these, and point mutants, to identify an oncogenic determinant of Ad 12 located to the spacer region between conserved regions 2 and 3 of the E1A gene. The specific aims of this program over the next five years are to: a. Continue phenotypic analysis of the Ad12-based viruses with chimeric E1A genes, and of rat and mouse cells transformed by these viruses, with a view to defining the potential influence of the E1A spacer region on immunological and other host-related factors which help to shape transformed cell tumorigenicity. b. Continue mutational analysis of the Ad12 E1A nonconserved spacer region which acts as a determinant of oncogenicity. c. Expand the chimeric approach to determine if other E1A regions are required for oncogenicity. d. Carry out phenotypic analysis of the Ad5-based chimeric viruses which carry the Ad12 spacer region and other regions, and determine if the spacer segment can act autonomously. e. Define the molecular basis for the role of the Ad12 E1A spacer region in oncogenic transformation. f.Assess the roles of the Ad5 and Ad12 E1B 55K proteins in oncogenicity. g. Determine if the products of viral genes outside the El transforming region influence the oncogenic capacity of Ad5 and Ad12. h. Explore the basis for the strong susceptibility of Hooded Lister rats to tumor induction by Ad12.

该研究计划的总体长期目标是使用 遗传，免疫学和分子方法来定义功能 5型和12型腺病毒（AD5和AD12）E1A和E1B蛋白， 也许还有其他病毒蛋白，在转化大鼠和小鼠细胞中发挥作用 体外，并调节发生在早期阶段的事件 人类细胞中的生产感染。一个特定的目标是了解 啮齿动物中AD12强的致癌性的基础 该血清型和非结构性类型之间的致癌性差异 5病毒。为此，我们已经构建并正在描述AD12和 基于AD5的病毒含有嵌合E1a基因，并使用了一些 这些和点突变体以识别AD 12的致癌决定因素 位于E1a的保守区域2和3之间的间隔区域 基因。在接下来的五年中，该计划的具体目的是： 一个。继续用嵌合E1a对基于AD12的病毒进行表型分析 基因，以及由这些病毒转化的大鼠和小鼠细胞的观点 定义E1A间隔区对潜在影响 免疫学和其他与宿主相关的因素，有助于塑造 转化的细胞肿瘤性。 b。继续对AD12 E1A未经保存的间隔区域的突变分析 它是致癌性的决定因素。 c。扩展嵌合方法以确定其他E1A区域是否是 致癌性所需。 d。对基于AD5的嵌合病毒进行表型分析 携带AD12垫片区域和其他区域，并确定垫片是否是 细分市场可以自主行动。 e。定义AD12 E1A间隔区域作用的分子基础 在致癌中。 f.ASSESS AD5和AD12 E1B 55K蛋白在致癌性中的作用。 g。确定EL转化之外的病毒基因的产物是否 区域影响AD5和AD12的致癌能力。 h。探索 带帽李斯特大鼠对肿瘤的强敏感性的基础 AD12诱导。