ANALYSIS OF THE SHOPE-PAPILLOMA CARCINOMA SYSTEM

SHOPE-乳头状瘤系统的分析

• 批准号: 3481799
• 负责人: FELIX O WETTSTEIN
• 金额: $ 17.9万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-06-30 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3481799
• 关键词: Shope papilloma; affinity chromatography; carcinoma; cell differentiation; clone cells; cocarcinogen; density gradient ultracentrifugation; gel electrophoresis; genetic manipulation; genetic translation; host neoplasm interaction; immunochemistry; immunofluorescence technique; molecular oncology; neoplasm /cancer immunology; neoplasm /cancer transplantation; rabbit papillomavirus; radiotracer; simian virus 40; viral carcinogenesis; virus DNA; virus RNA; virus antigen; virus genetics; virus related neoplasm /cancer; virus replication

We have mapped by S1 and ExoVII digestion and by primer extension viral transcripts in nonvirus producing domestic rabbit papillomas and carcinomas and in virus producing domestic rabbit papillomas and determined their coding capacity. The major transcripts can code for E7 and a truncated E6 protein, and minor ones were identified which can code for E2 and a full sized E6 protein. All early RNA is transcribed from promotors with TATA boxes. Late transcripts of 2.6 and 4.8 kb unique to virus producing tumors code for an L1 and an L2 or possibly an L2-L1 fusion protein. Both share the same leader exon and a capsite located in the untranslated region which is not preceded by a TATA box. This indicates that a switch from early to late transcription involves recognition of a new promotor and suppression of transcription termination at the early polyadenylation site. Expression of early proteins in COS-7 cells has permitted us to identify the E6 protein and preliminary data have also been obtained for E2. DNA analysis of HPV-16 in cervical cancer has shown that in a majority, the HPV-16 DNA is integrated. Some also contain plasmid DNA. Transcript mapping indicated that the major transcripts code for E7 and E6 and it suggests that only integrated DNA is functional and integration, at least of some DNA, may be required for cancer progression. The E7 protein of HPV-16 has been identified as a cytoplasmic phosphoprotein with a short half life. Cottontail rabbit papillomavirus provides the only animal model system and our long term objective is to understand the molecular biology of the virus and particularly how the virus contributes to the development of malignant tumors. In the present application, we propose to: 1) Identify and biochemically characterize ealy viral proteins with respect to subcellular distribution, modification, association with cellular proteins and possible enzymatic activity; 2) Determine their biological function through a genetic analysis in animals and epithelial cell culture; 3) Initiate experiments related to immunologic phenomena.

我们已经通过S1和ExoVII消化以及通过引物 非病毒产生家兔中的延伸病毒转录本 乳头状瘤和癌以及产病毒家兔 乳头状瘤，并确定其编码能力。 主要 转录本可以编码E7和截短的E6蛋白，以及次要的 其中一个可以编码E2和一个全尺寸的E6 蛋白 所有的早期RNA都是从具有TATA的启动子转录的 箱. 病毒特有的2.6和4.8 kb晚期转录物 产生编码L1和L2或可能L2-L1的肿瘤 融合蛋白 两者共享相同的前导外显子和一个帽位点 位于非翻译区，前面没有 塔塔盒子。 这表明从早到晚的转变 转录涉及识别新的启动子， 抑制转录终止在早期 多聚腺苷酸化位点。 COS-7细胞中早期蛋白的表达 使我们能够识别E6蛋白和初步数据， E2也是如此。 宫颈癌组织中HPV-16的DNA分析 宫颈癌已经表明，在大多数情况下，HPV-16 DNA是 整合 有些还含有质粒DNA。 转录作图 表明主要转录本编码E7和E6， 表明只有整合的DNA才有功能和整合， 至少某些DNA可能是癌症进展所必需的。 HPV-16的E7蛋白已被鉴定为细胞质内的 磷蛋白，半衰期短。 棉尾兔 乳头瘤病毒提供了唯一的动物模型系统， 长期目标是了解 病毒，特别是病毒如何有助于 恶性肿瘤的发展。 在本申请中，我们提出：1）识别和 对早期病毒蛋白进行生物化学表征 亚细胞分布、修饰、与细胞 蛋白质和可能的酶活性; 2）确定它们的 通过动物遗传分析的生物学功能， 上皮细胞培养; 3）启动与 免疫现象。