ACIDIFICATION OF HOST CELL ENDOSOMES & VIRUS RESISTANCE

宿主细胞内体的酸化

• 批准号: 3133339
• 负责人: WILLIAM S SLY
• 金额: $ 9.69万
• 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/3133339
• 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.

这项研究的主要目的是确定内体的作用， 病毒穿透宿主细胞时的酸化。 有五个具体的 目的： 1)我们认为，成功的病毒感染（病毒复制和 新病毒颗粒的繁殖）通常取决于 内化病毒通过酸性区室，内体。 我们有 发展了亚细胞分离技术， 分离的内体通过ATP依赖性机制酸化。 我们 第一个目的是确定内体的ATP依赖机制 酸化 2)我们已经从几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都具有“交叉抗性”。 初步 结果暗示了内体酸化的缺陷， “交叉抗性”。 我们的第二个目标是验证和表征 交叉抗性突变体中内体酸化缺陷。 3)已报道某些胺抑制对细胞凋亡的诱导。 抗病毒状态的干扰素。 我们的第三个目标是检验假设 胺的这种作用可以解释为干扰素需要 通过酸性隔室进入宿主细胞以诱导抗病毒抗体 状态 4)内体的Mg++和ATP依赖性酸化可能取决于 在Mg-ATPase上。 我们的第四个目标是确定负责 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，用于研究 内体酸化作为病毒感染的抑制剂， 和病毒感染抑制剂对核内体的作用 酸化 这些研究有助于阐明内体在病毒感染中的作用 渗透宿主细胞，并可能提供框架，开发新的 抗病毒化合物。