GENETICS OF THE TRANSCRIPTION PROCESS

真核RNA聚合酶III负责转录许多 小的RNA分子，包括所有细胞质的tRNA。初级识别 PolIII转录复合体对tRNA基因的特异性存在于 存在于染色质中但可以分离的两种或两种以上蛋白质 来自全细胞提取液中的RNA聚合酶III。形成一种 防止了tRNA基因和这些蛋白质之间的预启动复合体 通过改变二氢U和T Psi臂中的序列的突变 TRNA编码序列。本研究项目的目的是剖析 分离并识别在过程中发生的各种DNA-蛋白质相互作用 真核生物tRNA基因转录的启动和终止。 酵母中的OchRE抑制tRNA基因提供了独特的机会 分离和研究影响转录的tRNA基因的两个突变 以及与基因相互作用的蛋白质成分的基因突变 在转录过程中。可以预见，对这种类型的基因分析 影响酵母SUP4基因表达的突变共同作用 通过对受这些突变影响的蛋白质进行生化研究，威尔 提供对转录过程的新见解。因为这种方法是 发展起来，应该也适用于分子遗传学分析。 RNA聚合酶II，酵母中制造mRNA的酶和 其他真核生物。

Eukaryotic RNA polymerase III is responsible for the transcription of many small RNA molecules, including all cytoplasmic tRNAs. Primary recognition specificity of tRNA genes by the PolIII transcription complex resides in two or more proteins which are present in chromatin but can be separated from RNA polymerase III in whole cell extracts. Formation of a pre-initiation complex between a tRNA gene and these proteins is prevented by mutations which alter sequences within the Dihydro U and T Psi arms of tRNA coding sequence. The goal of this research project is to dissect apart and identify the various DNA-protein interactions which occur during the initiation and termination of transcription of eukaryotic tRNA genes. Ochre-suppressor tRNA genes in yeast provide unique opportunities to isolate and study both mutations in tRNA genes that affect transcription and mutations in genes for protein components that interact with the genes during transcription. It is envisioned that the genetic analysis of such mutations affecting expression of the yeast SUP4 tRNATyr gene, together with biochemical studies of the proteins affected by these mutations, will provide new insights into the transcription process. As this approach is developed, it should also be applicable to molecular genetic analysis of the function of RNA Polymerase II, the enzyme which makes mRNA in yeast and other eukaryotes.