RNA PROCESSING AND RIBONUCLEOPROTEIN

说明(改编自申请人的建议书)： 调查两个小RNA将被作为模型系统进行调查 进一步加深了我们对RNA流量和本地化的理解。这些研究 是这个实验室最近工作的合乎逻辑的延伸，在这个实验室中 活细胞中不同RNA的胞内运输通过 显微注射荧光RNA，结合并行定位 通过原位杂交和免疫细胞化学检测内源RNA。在……里面 目的1核糖核蛋白RNA亚基的核内转运 将研究核糖核酸酶P，重点是核仁相 它的生物合成、RNP组装和可以想象的功能。核糖核酸酶P的突变体 将通过注射荧光RNA和通过表达来研究 标记基因来定义核仁靶向信号。这些研究将 通过分析RNP组装中涉及的序列来补充，使用 新近研制出的抗体。核糖核酸酶P在体内发挥作用的可能性 核糖体前RNA加工或其他核仁RNA加工将 在非洲爪哇卵母细胞中进行研究，如果有必要，也在哺乳动物细胞中进行研究。在……里面 目标2申请人最近发现的信号识别粒子 (SRP)RNA定位(至少是暂时的)在核仁中将是 搜索本地化所需的序列的基础。相同 研究应提供有关核出口所需序列的信息。 这两个相互关联的项目检验了核仁是一种 比之前想象的更具多功能的细胞器。

DESCRIPTION (Adapted from applicant's proposal): In the proposed investigation two small RNAs will be investigated as model systems for furthering our understanding of RNA traffic and localization. These studies are a logical extension of recent work from this laboratory in which the intracellular traffic of various RNAs in living cells was investigated by microinjection of fluorescent RNAs, coupled with parallel localization of the endogenous RNAs by in situ hybridization and immunocytochemistry. In Aim 1 the intranuclear traffic of the RNA subunit of the ribonucleoprotein enzyme RNase P will be investigated, with emphasis on the nucleolar phase of its biosynthesis, RNP assembly and conceivably function. Mutants of RNase P RNA will be investigated by injection of fluorescent RNA and by expression of marked genes to define nucleolar targeting signals. These studies will be complemented by analysis of the sequences involved in RNP assembly, using recently developed antibodies. The possibility that RNase P functions in pre-ribosomal RNA processing or other nucleolar RNA processing will be investigated in Xenopus oocytes, and if warranted in mammalian cells. In Aim 2 the applicant's recent discovery that signal recognition particle (SRP) RNA is localized (at least transiently) in the nucleolus will be the basis for a search for sequences required for this localization. The same studies should provide information on sequences required for nuclear export. These two inter-related projects test the hypothesis that the nucleolus is a more pluri-functional organelle than previously thought.