STRUCTURE/EXPRESSION OF YEAST VIRAL DOUBLE-STRANDED RNAS

酵母病毒双链RNA的结构/表达

• 批准号: 3271003
• 负责人: JEREMY A BRUENN
• 金额: $ 14.87万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-05-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3271003
• 关键词: DNA directed RNA polymerase; RNA; RNA biosynthesis; RNA virus; Saccharomyces; chemical fingerprinting; density gradient ultracentrifugation; electron microscopy; fatty acid binding protein; fungal genetics; gel electrophoresis; genetic manipulation; genetic mapping; genetic regulation; genetic transcription; glycoproteins; immunochemistry; messenger RNA; molecular cloning; mutant; nucleic acid hybridization; nucleic acid sequence; radiotracer; scintillation counter; thin layer chromatography; transcription termination; tritium; virus RNA; viruslike particle

ScV is a simple ds RNA virus with two, separately encapsidated dsRNAs. Like other fungal viruses, ScV particles are communicated from cell to cell only by mating. The larger viral dsRNA (L) encodes the major capsid protein. The smaller (M) encodes a toxin lethal to strains without ScV-M particles. INternal deletions of M result in defective-interfering (DI) particles containing fragments of M (ScV-S particles). Viral particles have a transcriptase activity. There are many unresolved questions about the ScV system that we hope to address in this project. A completed sequence analysis of L and M, combined with immunological screening for peptides predicted by sequence analysis should complete the catalog of viral proteins. An exact mapping of the in vivo and in vitro viral mRNAs will complement this analysis. There is no definitive evidence on the mechanism of transcription and replication of the viral dsRNAs. With the appropriate use of cDNAs cloned in the single-stranded phage M13, it should be possible to distinguish between conservative and semiconservative transcription in vitro. Suppression remains a useful phenomenon for understanding both viral interactions and virus-host interactions, since DI particles displace their parental virus by competing for some product present in limiting amount. The DI RNAs themselves will provide useful information about the viral transcriptase: a comparison of S dsRNAs should help us deduce signals for transcription termination and initiation. Combined with a complete analysis of the coding regions of M and similar sequence analysis of L, this comparison of S dsRNAs should define sites necessary for transcription initiation, replication, and packaging. Exact definition of such sites may become possible by construction of cDNA clones capable of expressing the complete viral plus strand in transformed yeast cells without indiginous virus, thus resurrecting the virus.

SCV是一种简单的DS RNA病毒，具有两个分别封装的DSRNA。 像其他真菌病毒一样，SCV颗粒从细胞到细胞传达 仅通过交配。 较大的病毒dsRNA（L）编码主要的衣壳 蛋白质。 较小的（M）编码毒素致命为没有SCV-M的菌株 颗粒。 M的内部缺失导致干扰不良（DI） 包含M（SCV-S颗粒）片段的颗粒。 病毒颗粒 具有转录酶活性。 关于 我们希望在该项目中解决的SCV系统。 一个完工 L和M的序列分析，结合免疫筛查 序列分析预测的肽应完成 病毒蛋白。 体内和体外病毒mRNA的精确映射 将补充这一分析。 没有关于 病毒DSRNA的转录和复制机制。 与 适当使用克隆单链噬菌体M13中的cDNA，应该 可以区分保守和半保守 体外转录。 抑制仍然是一个有用的现象 了解病毒相互作用和病毒宿主相互作用，因为DI 颗粒通过竞争某些产品来取代其父母病毒 以限制数量存在。 di rnas本身将提供有用的 有关病毒转录酶的信息：S DSRNA的比较应 帮助我们推断出转录终止和启动的信号。 结合对M和类似编码区的完整分析 l的序列分析，S dsrNA的这种比较应定义位点 转录启动，复制和包装所必需的。 精确的 通过构建cDNA克隆，可能会成为这种位点的定义 能够在转化的酵母中表达完整的病毒和链 没有土著病毒的细胞，从而使病毒复活。