ACIDIFICATION OF HOST CELL ENDOSOMES & VIRUS RESISTANCE

宿主细胞内体的酸化

• 批准号: 3133338
• 负责人: WILLIAM S SLY
• 金额: $ 10.58万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-09-30 至 1987-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3133338
• 关键词: adenosine triphosphate; adenosinetriphosphatase; amines; antiviral agents; density gradient ultracentrifugation; diphtheria toxin; fluorescent dye /probe; interferons; kidney cell; lysosomes; mutant; organelles; photochemistry; pinocytosis; radiotracer; virus infection mechanism; virus replication; viruslike particle

The broad aim of this research is to define the role of endosome acidification in virus penetration of host cells. There are five specific aims: 1) We believe that successful virus infection (viral replication and propagation of new virus particles) normally depends on passage of internalized virus through an acidic compartment, the endosome. We have developed subcellular fractionation techniques which allowed us to show that the isolated endosome acidifies by an ATP-dependent mechanism. Our first aim is to define the ATP-dependent mechanism of endosome acidification. 2) We have obtained, from several sources, Mammalian cell mutants that are "cross resistant" to both diphtheria toxin and animal viruses. Preliminary results implicate a defect in endosome acidification as the basis for "cross resistance". Our second aim is to verify and characterize the defect in acidification of endosomes in cross resistant mutants. 3) Certain amines have been reported to inhibit the induction of the anti-viral state by interferon. Our third aim is to test the hypothesis that this effect of amines is explained by a requirement for interferon to enter the host cell through an acidic compartment to induce the anti-viral state. 4) The Mg++ and ATP-dependent acidification of endosomes presumably depends on a Mg-ATPase. Our fourth aim is to identify the ATPase responsible for acidification of endosomes, and to purify and characterize it. 5) Our fifth aim is to develop a model system for studying inhibitors of endosome acidification for their action as inhibitors of virus infection, and inhibitors of virus infection for their effects on endosome acidification. These studies should elucidate the role of the endosome in virus penetration of host cells and may provide the framework for developing new anti-viral compounds.

这项研究的主要目标是确定内体的作用 病毒渗透宿主细胞的酸化。 具体有五项 目标： 1）我们相信成功的病毒感染（病毒复制和 新病毒颗粒的传播）通常取决于 病毒通过酸性区室（内体）内化。 我们有 开发了亚细胞分离技术，使我们能够展示 分离的内体通过 ATP 依赖性机制酸化。 我们的 第一个目标是确定内体的 ATP 依赖性机制 酸化。 2）我们从多个来源获得了哺乳动物细胞突变体 对白喉毒素和动物病毒具有“交叉抗性”。 初步的 结果表明内体酸化缺陷是 “交叉抵抗”。 我们的第二个目标是验证和表征 交叉耐药突变体中内体酸化缺陷。 3) 据报道，某些胺会抑制诱导 干扰素的抗病毒状态。 我们的第三个目标是检验假设 胺的这种作用可以通过干扰素的需要来解释 通过酸性隔室进入宿主细胞以诱导抗病毒 状态。 4) 内体的 Mg++ 和 ATP 依赖性酸化可能取决于 在 Mg-ATP 酶上。 我们的第四个目标是确定负责的 ATPase 内体的酸化，并对其进行纯化和表征。 5）我们的第五个目标是开发一个模型系统来研究抑制剂 内体酸化作为病毒感染抑制剂的作用， 和病毒感染抑制剂对内体的影响 酸化。 这些研究应该阐明内体在病毒中的作用 渗透宿主细胞并可能为开发新的 抗病毒化合物。