MUTATIONS AFFECTING THE PRODUCTION OF MATURE RNAS

影响成熟 RNA 产生的突变

• 批准号: 3275164
• 负责人: Anita K Hopper
• 金额: $ 21万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-09-01 至 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3275164
• 关键词: RNA biosynthesis; Saccharomyces; eukaryote; fungal genetics; gene expression; genetic transcription; laboratory rabbit; methylation; nucleic acid sequence; polymerase chain reaction; ribosomal RNA; transfer RNA

The goal of the proposed research is to gain an understanding of the steps of eukaryotic pre-tRNA and pre-rRNA maturation and the gene products that play a role in these processes. In all organisms initial transcripts can differ significantly from the mature RNAs that function in protein synthesis. Although there is a general outling of the steps necessary to generate mature tRNAs and rRNAs, few of the enzymes that catalyze nucleolytic, splicing and modification reactions have been characterized and even fewer of the genes coding and regulating these activities have been studied. Furthermore, it is known that pre-rRNA processing steps occur upon ribonuclear particles and it is possible that such particles participate in pre-tRNA processing. Thus, gene products other than those responsible for the catalytic activities are also likely to be required for processing reactions. We are utilizing genetic, molecular and biochemical approaches to answer four questions regarding the processing of pre-tRNA and pre-rRNAs in Saccharomyces cerevisiae: (1) What parts of pre- tRNAs are important to the processing machinery? We will study the in vivo consequences upon pre-tRNA processing of single mutations in a gene encoding tRNATyr; (2) What products are involved in pre-tRNA processing? We are attempting to identify, by mutation or by gene amplification, genes that affect the maturation of tRNA; (3) Is pre-tRNA processing associated with ancillary superstructural proteins? (4) What are the functions of the genes that previously had been shown to affect pre-tRNA and pre-rRNA processing? We have now identified 6 genes (RNA1, SRN1, LOS1, pC6, RRP1 and SRD1) that affect processing. Surprisingly, it does not appear that any of these code for known catalytic activities and therefore these genes may serve to identify products that were unanticipated from the biochemical pathways of the processing reactions. Our approach to study these genes is to clone the wild-type genes, to locate the gene products within yeast, to biochemically characterize these products, and to identify other genes with related functions.

拟议的研究的目的是了解 真核pre-tRNA和前rNNA成熟的步骤以及 在这些过程中发挥作用的基因产品。 总的来说 有机体初始成绩单可能与 成熟的RNA在蛋白质合成中起作用。 虽然那里 是产生成熟所需的步骤的一般范围 trnas和rRNA，很少催化核酸化的酶 剪接和修改反应已被表征，并且 编码和调节这些活动的基因甚至更少 已经被研究了。 此外，众所周知 核糖核颗粒上发生处理步骤，可能 这样的颗粒参与了tRNA前处理。 因此， 除催化性的基因产品以外 处理反应也可能需要活动。 我们正在利用遗传，分子和生化方法 回答有关pre-tRNA的处理的四个问题 酿酒酵母中的葡萄球菌前：（1） TRNA对处理机械很重要？ 我们将学习 体内对单个单一处理的后果 编码trnatyr的基因中的突变； （2）什么是产品 参与tRNA处理？ 我们试图识别， 通过突变或通过基因扩增，影响该基因 tRNA的成熟； （3）是与 辅助上层结构蛋白？ （4） 以前已证明会影响tRNA的基因和 前rNNA处理？ 我们现在已经鉴定了6个基因（RNA1， 影响处理的SRN1，LOS1，PC6，RRP1和SRD1）。 令人惊讶的是，似乎没有任何这些代码 催化活性，因此这些基因可能有助于 识别从生化中意外的产品 处理反应的途径。 我们学习的方法 这些基因是克隆野生型基因，以定位基因 酵母中的产品，以生化为特征 产品，并确定具有相关功能的其他基因。