MEASLES VIRUS: THE VIRION AND ITS REPLICATIVE STRATEGY

麻疹病毒：病毒粒子及其复制策略

• 批准号: 3130257
• 负责人: Stephen A. Udem
• 金额: $ 21.59万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3130257
• 关键词: HeLa cells; autoradiography; binding proteins; gel electrophoresis; gene expression; genetic transcription; genome; immunoprecipitation; laboratory rabbit; measles; microorganism reproduction; molecular cloning; molecular pathology; nucleic acid probes; nucleic acid sequence; protein biosynthesis; radiotracer; ribonucleoproteins; subacute sclerosing panencephalitis; tissue /cell culture; virulence; virus RNA; virus envelope; virus infection mechanism; virus morphology; virus protein; virus replication

The aims of this proposal remain the detailed characterization of (1) the mechanisms by which efficient reproduction of measles virus is achieved and (2) the ways in which these pathways of replication are altered in the evolution of persistent measles virus infections, in order, ultimately, to (3) gain insight into the role played by such persistent infection in the pathogenesis of chronic human disease. Despite recent progress in defining the sequence, structure, and organization of the measles virus genome, the pathways by which transcription and genomic replication are accomplished remain vague and uncertain. Particular attention will be given to these, as yet undefined, features of viral reproduction by exploiting systems and preliminary observations recently generated by this laboratory, including: (1) an in vitro system able to accomplish transcription only when supplemented by a host cell factor -- a system in which the role(s) played by virus-specified and by host cell proteins can now be dissected and reconstructed, and (2) a membrane blot-hybridization system showing binding of measles virus genome and anti-genome RNA to a approximately 90 kD. host-cell protein --a method by which the polarity and sequences of viral RNA responsible for this host-cell protein:nucleic acid interaction can be analyzed and perhaps by which the possible significance of this interaction to transcription and/or replication can be assessed. In addition, using our extensively studied SSPE cell line (IP-3-Ca), efforts to identify the mechanism(s) by which measles virus matrix protein expression is restricted in the persistently infected brain cells of patients with SSPE will continue. The major aim now is to clone the matrix genes of the IP-3-Ca cell and of the genome of a revertent SSPE virus that emerged from this cell line, and to subject these cDNAs to sequence analysis in order to evaluate the hypothesis that genomic mutations accumulated in the course of measles virus persistence ultimately determine both the nature of and the level at which matrix protein expression is restricted in SSPE.

该提案的目的仍然是 （1）有效繁殖麻疹的机制 获得病毒，（2）这些途径的方式 持续麻疹病毒的演变改变了复制 最终，感染最终（3）了解角色 通过这种持续感染在慢性发病机理中发挥的作用 人类疾病。 尽管最近在定义序列，结构和 麻疹病毒基因组的组织，该途径 转录和基因组复制保持了 含糊和不确定。 将特别注意这些， 尚未定义，通过利用病毒复制的特征 系统和初步观察最近产生了 实验室，包括：（1）一个能够完成的体外系统 仅在补充宿主细胞因子时转录 - a 病毒指定和宿主扮演的角色的系统 现在可以解剖和重建细胞蛋白，（2）A 膜印迹杂交系统显示麻疹的结合 病毒基因组和抗基因组RNA至约90 kD。 宿主细胞蛋白 - 极性和序列的方法 负责此宿主细胞蛋白的病毒RNA：核酸 可以分析互动，也许可以通过 这种相互作用对转录和/或复制的重要性 可以评估。 另外，使用我们的广泛研究的SSPE细胞系（IP-3-CA）， 确定麻疹病毒的机制的努力 基质蛋白表达受到持续感染的限制 SSPE患者的脑细胞将继续。 主要目标 现在是克隆IP-3-CA细胞的基因基因和 从该细胞中出现的恢复SSPE病毒的基因组 行，并对这些cDNA进行序列分析，以便 评估基因组突变积累的假设 麻疹病毒持久性的过程最终决定 基质蛋白表达的性质和水平是 限制在SSPE中。