BIOSYNTHESIS OF RNAS

RNAS的生物合成

• 批准号: 3270256
• 负责人: CHRISTINE GUTHRIE
• 金额: $ 16.5万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-02-01 至 1990-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3270256
• 关键词: alleles; chromatography; endonuclease; eukaryote; fungal genetics; gel electrophoresis; gene expression; genetic manipulation; genetic transcription; lethal genes; messenger RNA; mutant; nucleic acid sequence; prokaryote; radiotracer; transfer RNA; ultraviolet spectrometry

Despite intensive study in numerous laboratories, our understanding of the molecular mechanisms which specify the accurate and efficient splicing of intron-containing mRNA nuclear transcripts has progressed only slightly beyond the phenomenological. The same is true for other steps in the maturation of messages, including the generation of 3' termini and polyadenylation. Attention has recently been drawn to a group of six small nuclear RNAs (snRNAs), whose extraordinary structural conservation, together with a variety of circumstantial evidence, suggest their participation in functionally conserved steps in eukaryotic gene expression. For example, U1 RNA is hypothesized to mediate intron removal via complementarity to conserved sequences at the splice junction(s). Because of the lack of tractable experimental systems among the higher eukaryotes, these hypotheses have yet to be critically tested. We propose to exploit the powerful genetic techniques uniquely available in yeast to achieve these ends. We are employing two complementary strategies. The first exploits our recent demonstration of the existence in Saccharomyces cerevisiae of snRNAs encoded by single copy genes; this allows us to perform definitive tests of these models by analyzing the consequences of null and conditional alleles in each of these genes. Moreover, the isolation of extragenic suppressors of these mutations will identify cellular components which interact with the snRNAs; these should include the snRNP proteins, as well as potential "target" RNAs: the pre-mRNA substrates. Our second approach takes advantage of our recent identification of a mutant 5' splice junction in a yeast mRNA intron. In this case, the ability to isolate extragenic suppressors should allow a direct test of the "Ul guide model" for RNA splicing

尽管对众多实验室进行了深入研究，但我们对 分子机制，这些机制指定了准确有效的剪接 含有内含子的mRNA核转录本仅略有进展 超越现象学。 对于其他步骤也是如此 消息的成熟，包括3'末端和 聚腺苷酸化。 最近注意到一组六个小型 核RNA（SNRNA），其非凡的结构保护， 以及各种间接证据，表明他们 参与真核基因的功能保守步骤 表达。 例如，假设U1 RNA介导内含子去除 通过与剪接连接处的保守序列的互补性。 由于较高的实验系统缺乏可行的实验系统 真核生物，这些假设尚未经过严格检验。 我们建议 利用酵母中独特可用的强大遗传技术 达到这些目的。 我们采用两种互补策略。 第一个利用我们的 最近证明了Snrnas酿酒酵母中存在的存在 由单复制基因编码；这使我们能够执行确定的测试 这些模型通过分析零和条件等位基因的后果 在每个基因中。 此外，隔离基因抑制器 这些突变中将确定与之相互作用的细胞成分 snrnas;这些应包括SNRNP蛋白质以及潜力 “目标” RNA：前MRNA底物。 我们的第二种方法采用 我们最近识别突变体5'剪接连接的优势 酵母mRNA内含子。 在这种情况下，隔离基因的能力 抑制器应允许直接测试RNA的“ UL指南模型” 剪接