SV40 CHROMOSOME: T-ANTIGEN COMPLEXES AND VIRAL FUNCTION

SV40 染色体：T 抗原复合物和病毒功能

• 批准号: 3174758
• 负责人: LOIS C TACK
• 金额: $ 8.47万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-08-01 至 1986-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3174758
• 关键词: chromatin; electron microscopy; gel electrophoresis; gene expression; genetic transcription; histocompatibility antigens; monoclonal antibody; radiotracer; simian virus 40; thiols; tissue /cell culture; tumor antigens; virus DNA; virus antigen; virus genetics; virus infection mechanism

The genetic information of all higher organisms resides in their chromosomes; therefore, the molecular mechanisms by which chromosomes are replicated and transcribed are fundamental to our understanding of the proliferation and differentiation of all cells. Simian virus 40 chromosomes utilize cellular host processes to accomplish these functions and thus, are apropriate but relatively simple model systems for study. The properties of virally expressed proteins as they relate to the control of viral chromosome functions are also of interest. This proposal considers the question of how viral chromatin structure, specifically T-antigen: SV40 chromosome complexes, afffects the replication and expression of its genetic information. The use of immunoprecipitation techniques utilizing several monoclonal anti-T antibodies to purify T-containing mature, replicating and transcribing SV40 chromosomes, in vivo and in vitro replication and transcription assays to characterize function along with techniques such as electron microscopy, restriction endonuclease digestion, hybridization assays, sedimentation fractionation and gel electrophoresis to identify, characterize and quantitate T-containing DNA species will clarify the role of T-antigen with respect to in vivo SV40 chromosomal replication and transcription. Nuclease digestion and specific labeling studies of T-containing viral chromosomes will be used to determine whether chromatin phasing directs biological events at specific DNA sequences. In addition, the chemistry and functional properties of a highly reactive, aminesensitive ester site in SV40 T-antigen will be examined with respect to viral function. Similar sites recently described in several plasma proteins are not only critical for function of the native molecules, but are, upon activation, capable of covalent interaction with receptor molecules.

所有高等生物的遗传信息都存在于 染色体;因此，染色体的分子机制 复制和转录是我们理解 所有细胞的增殖和分化。 猿猴病毒40 染色体利用细胞宿主过程来完成这些功能 因此，是适当但相对简单的研究模型系统。 病毒表达的蛋白质的性质，因为它们与对照 病毒染色体功能的研究也很有意义。 这项建议 考虑了病毒染色质结构的问题，特别是 T抗原：SV 40染色体复合物，影响复制和 表达其遗传信息。 免疫沉淀的应用 利用几种单克隆抗T抗体的技术， 体内含T成熟、复制和转录SV 40染色体 以及体外复制和转录测定来表征功能 沿着技术如电子显微镜、限制性内切酶 消化、杂交测定、沉降分级分离和凝胶 电泳以鉴定、表征和定量含T DNA 物种将阐明T抗原对于体内SV 40的作用 染色体复制和转录。 核酸酶消化和特异性 含T病毒染色体的标记研究将用于 确定染色质定相是否在特定的时间点指导生物学事件 DNA序列 此外，还研究了一种新型纳米复合材料的化学和功能特性。 SV 40 T抗原中的高反应性、胺敏感性酯位点将被 检查病毒功能。 类似的网站最近描述 在几种血浆蛋白质中，不仅对天然蛋白的功能至关重要， 分子，但在活化时，能够与 受体分子