Structure and Function of Self-replicate RNAs

自我复制RNA的结构和功能

基本信息

批准号：
59380025
负责人：
OKADA Yoshimi
金额：
$ 15.74万
依托单位：
University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Co-operative Research (A)
财政年份：
1984
资助国家：
日本
起止时间：
1984 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-59380025/
关键词：
RNA Viral RNA Viroid Replication of RNA Genome structure cDNAクローン

项目摘要

An in vitro transcription system to produce infectious TMV RNA from a cloned cDNA has been established. Using this system, several mutations were introduced at or near the termination codon of the 130K protein. The 130K and 180K proteins are confirmed to be involved in viral RNA replication. The pathogenisities and nucleotide sequences of cucumber pale fruit viroid, grapevine viroid and hop stunt viroid was investigated and demonstrated that the pathogenisities of these three viroids are quite simialr, though some differences are recognized in their nucleotide sequences. Nucleotide sequence of apple scar skin viroid has been determined.Multiple functions of the RNA-dependent RNA polymerase in influenza virus have been identified including the endonucleolvtic cleavage of capped RNA, the primer dependent RNA synthesis, the polyadenylation of transcripts and an apparent proofreading function. For studying the antigenic determinant sites or functional domain of sialic acid binding activity of influenza virus HA, site directed mutagenesis was applied. The genome organization of Sendai virus was elucidated completely and it was found that the nonstructural C protein of the virus plays an important role in induction of interferon. In vitro modified polioviruses were constructed by using the infectious cDNA clone. Biological tests including plaque size marker test and monkey neurovirulence test were performed on these mutant viruses. The results suggested that interactions between different genome loci of the RNA were importantly involved in the replication and gene expression.The complete nucleotide sequence of the RNA coliphage GA and SP were determined. The amino acid residues in the coat protein causing the changes of the serological properties from GA to MS2 were identified.

已经建立了从克隆的 cDNA 产生感染性 TMV RNA 的体外转录系统。使用该系统，在 130K 蛋白的终止密码子处或附近引入了多个突变。 130K和180K蛋白被证实参与病毒RNA复制。对黄瓜淡果类病毒、葡萄类病毒和啤酒花矮化类病毒的致病性和核苷酸序列进行了研究，结果表明，这三种类病毒的致病性非常相似，尽管它们的核苷酸序列存在一些差异。苹果疤痕类病毒的核苷酸序列已确定。流感病毒中RNA依赖性RNA聚合酶的多种功能已被鉴定，包括加帽RNA的核酸内切、引物依赖性RNA合成、转录物的多腺苷酸化和明显的校对功能。为了研究流感病毒HA的唾液酸结合活性的抗原决定簇位点或功能域，应用了定点诱变。仙台病毒的基因组结构被完全阐明，发现该病毒的非结构C蛋白在诱导干扰素中发挥重要作用。使用感染性cDNA克隆构建体外修饰的脊髓灰质炎病毒。对这些突变病毒进行了生物学测试，包括噬斑大小标记测试和猴神经毒力测试。结果表明RNA不同基因组位点之间的相互作用在复制和基因表达中发挥着重要作用。确定了RNA大肠杆菌噬菌体GA和SP的完整核苷酸序列。鉴定了外壳蛋白中引起血清学特性从GA到MS2变化的氨基酸残基。