RNA-RNA ANNEALING PROTEINS

本研究的目的是分离和鉴定HeLa中的蛋白质 促进互补RNA之间碱基配对的细胞核 分子。在反义RNA的实验过程中 作为前mRNA剪接的探针，核提取液中存在的蛋白质 极大地加快了RNA-RNA的退火率。 这里概述了实验以确定其机制和特异性。 以及这些蛋白质在细胞中所起的作用。核子 萃取物将通过层析程序进行分级，以确定和 提纯促进RNA/RNA退变的蛋白质。化验方法将是 建立了针对不同类型的退火反应。纯净退火法 蛋白质将根据它们与RNA的相互作用来表征 以及促进碱基配对的能力。单抗将会是 针对退火蛋白制备的，用于表征细胞内 本地化和与RNA的关联是理解的第一步 这些蛋白质的生物学作用。这项工作对 了解介导RNA-RNA碱基配对的蛋白质的性质 活着。因此，这项研究将为进一步深入了解分子 高等生物中调控基因表达的相互作用及其相互作用 并增强我们在体内调节这些相互作用的能力 在试管中。

The goal of this research is to isolate and characterize proteins in HeLa cell nucleus which promote base-pairing between complementary RNA molecules. During the course of experiments in which antisense RNA was used as a probe for pre-mRNA splicing, proteins present in nuclear extracts were observed to greatly accelerate the rate of RNA-RNA annealing. Experiments are outlined here to determine the mechanism and specificity of annealing and the role that these proteins play in the cell. Nuclear extracts will be fractionated by chromatographic procedures to identify and purify proteins which promote RNA/RNA annealing. Assay methods will be established for different types of annealing reactions. Purified annealing proteins will be characterized with respect to their interactions with RNA and ability to promote base-pairing. Monoclonal antibodies will be prepared against annealing proteins and used to characterize intracellular localization and association with RNA as a first step towards understanding the biological role of these proteins. This work is important for understanding properties of proteins which mediate RNA-RNA base-pairing in vivo. This research will thus provide further insight into molecular interactions which regulate gene expression in higher organisms and their viruses and enhance our ability to modulate these interactions in vivo and in vitro.