SV40 ANTIGEN, CHROMATIN STRUCTURE AND VIRAL FUNCTION

SV40 抗原、染色质结构和病毒功能

• 批准号: 3174755
• 负责人: LOIS C TACK
• 金额: $ 14.5万
• 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 1989-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3174755
• 关键词: chromatin; electron microscopy; gel electrophoresis; gene expression; genetic mapping; genetic transcription; histocompatibility antigens; monoclonal antibody; mutant; radiotracer; simian virus 40; temperature sensitive mutant; thiols; tissue /cell culture; tumor antigens; virus DNA; virus antigen; virus genetics; virus infection mechanism; virus replication

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 understanding the proliferation and differentiation of all cells. Simian virus 40 (SV40) chromosomes utilize normal cellular host processes for viral DNA replication and transcription within permissive cells and thus are appropriate and relatively simple model systems for studying these processes. Analyzing the properties of virally-expressed proteins as they regulate viral transcription and replication during productive infection of eukaryotic cells is necessary for understanding how viruses function within host cells. This proposal is concerned with further studies on the structure and function of large T antigen, both cytoplasmic and chromosome-bound species, synthesized during SV40 lytic infection. Twenty-one monoclonal antibodies, specific for T, will be used to analyze the role of T Ag in each of the several steps required for viral DNA synthesis. These antibodies, recognizing different structural and functional sites as well as active and inactive subclasses of T, will permit a comprehensive analysis of the multifunctional T protein as it participates in a wide variety of activities related to replicative function. An amine-sensitive ester site in SV40 T is important for early steps in viral replication. Its replicative role will be analyzed using several defined SV40 mutants. This highly reactive ester bond appears related to thiolester sites in the complement gene family and is present in all papovavirus T Ags. Highly homologous sequences are also found in several other viral and cellular proteins. Using biochemical and genetic studies we will characterize and identify the SV40 trans acting chromatin-bound factor responsible for activating late viral gene transcription during lytic infection, concomitant with activation of viral replication.

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 复制和转录是理解 所有细胞的增殖和分化。 猴病毒40（SV 40） 染色体利用病毒DNA的正常细胞宿主过程 在允许细胞内复制和转录， 适当的和相对简单的模型系统来研究这些 流程. 分析病毒表达蛋白质的特性， 在生产性感染期间调节病毒转录和复制， 真核细胞是了解病毒如何在体内发挥作用所必需的。 宿主细胞 这项建议涉及进一步研究 大T抗原结构和功能 染色体结合的物质，在SV 40裂解感染期间合成。 21种T特异性单克隆抗体将用于分析 T抗原在病毒DNA所需的几个步骤中的每一个中的作用 合成. 这些抗体，识别不同的结构和 功能位点以及T的活性和非活性亚类，将 允许全面分析多功能T蛋白，因为它 参加各种各样的活动，有关复制 功能 SV 40 T中的胺敏感性酯位点对于早期免疫应答是重要的。 病毒复制的步骤。 它的复制作用将使用 几种确定的SV 40突变体 这种高活性的酯键 与补体基因家族中的硫酯位点相关，并存在于 所有乳多空病毒T抗原。 高度同源的序列也被发现， 其他几种病毒和细胞蛋白质。 利用生物化学和遗传学 研究，我们将表征和鉴定SV 40反式作用 负责激活晚期病毒基因的染色质结合因子 在裂解感染过程中转录，伴随着病毒的激活， 复制的