MUTATIONS AFFECTING THE PRODUCTION OF MATURE RNAS

建议对其调控、合成和表达的研究 真核tRNA可以在酵母中完成--一种简单的真核生物，在其中 可以进行遗传和生化相结合的研究。这项研究 旨在分离突变体或建立tRNA 合成或表达异常。实现这一目标的两种方法是 描述：(1)条件丧失能力的突变体的分离 抑制无意义的突变，也无法在不允许的情况下成长 温度(导致细胞失去抑制子的突变类型 活性将通过重建携带突变的抑制子菌株来研究 在影响酵母细胞生长的已知方面的基因中)； 用功能性RNA聚合酶III自杀细胞。突变体将是 对基因进行分析以确定互补基团的数量并 确定突变是否会影响一种以上的tRNA。核糖核酸 将对细胞进行生化分析，以建立一条 处理事件，以确定修饰碱基在tRNA功能中的作用，以及 研究RNA聚合酶III的亚基。

It is proposed that a study of the regulation, synthesis, and expression of eukaryotic tRNA can be accomplished in yeast - a simple eukaryote in which combined genetic and biochemical investigations can be conducted. The research is directed towards isolating mutants or establishing conditions for which tRNA synthesis or expression is aberrant. Two approaches to accomplishing this are described: (1) isolation of mutants which conditionally lose the ability to suppress nonsense mutations and also fail to grow at the nonpermissive temperature (the types of mutations which cause cells to lose suppressor activity will be studied by reconstructing suppressor stains bearing mutations in genes affecting well described aspects of yeast cells growth); (2) tritium suicide of cells with functional RNA polymerase III. The mutants will be analyzed genetically to determine the number of complementation groups and to determine whether the mutations affect more than one species of tRNA. The RNA from the cells will be analyzed biochemically to establish a pathway of processing events, to determine the role of modified bases in tRNA function, and to study the subunits of RNA polymerase III.