ANALYSIS OF THE SHOPE-PAPILLOMA CARCINOMA SYSTEM

SHOPE-乳头状瘤系统的分析

• 批准号: 3481801
• 负责人: FELIX O WETTSTEIN
• 金额: $ 18.22万
• 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/3481801
• 关键词: Shope papilloma; affinity chromatography; carcinoma; cell differentiation; chimeric proteins; clone cells; cocarcinogen; density gradient ultracentrifugation; gel electrophoresis; genetic manipulation; genetic translation; host neoplasm interaction; immunochemistry; immunofluorescence technique; laboratory rabbit; molecular oncology; neoplasm /cancer immunology; neoplasm /cancer transplantation; nucleic acid hybridization; protein biosynthesis; protein structure; rabbit papillomavirus; radiotracer; simian virus 40; viral carcinogenesis; virus DNA; virus RNA; virus antigen; virus genetics; virus protein; 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酶切和引物进行了定位 无病毒家兔体内延伸病毒转录本的研究 家兔乳头状瘤、癌及病毒的产生 并测定它们的编码能力。少校 转录本可以编码E7和截短的E6蛋白，以及次要的 鉴定出了可以编码E2和全尺寸E6的基因 蛋白。所有早期的rna都是从带有tata的启动子转录而来的。 盒子。病毒特有的2.6和4.8kb的晚期转录本 产生L1和L2或可能的L2-L1的肿瘤编码 融合蛋白。这两个基因都有相同的先导外显子和一个CAPITE 位于未翻译区域中，该区域前面没有 塔塔箱。这表明从早到晚的切换 转录包括识别新的启动子和 转录终止在早期的抑制 多聚腺苷酸化位点。早期蛋白在COS-7细胞中的表达 使我们能够鉴定E6蛋白和初步数据 也获得了E2。人乳头瘤病毒16型DNA分析 宫颈癌已经表明，在大多数情况下，HPV-16DNA是 集成的。有些还含有质粒DNA。成绩单映射 表明E7和E6的主要转录本编码以及它 表明只有整合的DNA才是功能和整合的， 至少是一些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 半衰期短的磷蛋白。棉尾兔 乳头瘤病毒提供了唯一的动物模型系统和我们的 长期的目标是了解人类免疫缺陷病毒的分子生物学。 病毒，特别是病毒如何对 恶性肿瘤的发展。 在本申请中，我们建议：1)识别和 伊利病毒蛋白的生物化学特征 亚细胞分布、修饰、与细胞的关联 蛋白质和可能的酶活性；2)确定它们的 通过对动物和动物的遗传分析来实现生物学功能 上皮细胞培养；3)启动与以下相关的实验 免疫现象。