RNA-RNA ANNEALING PROTEINS

本研究的目的是分离和表征HeLa中的蛋白质 促进互补RNA之间碱基配对的细胞核 分子。 在使用反义RNA的实验过程中， 用作前mRNA剪接探针，存在于核提取物中的蛋白质 观察到大大加速RNA-RNA退火的速率。 本文概述了实验以确定 退火和这些蛋白质在细胞中的作用。 核 将通过色谱程序对浸提液进行分级， 纯化促进RNA/RNA退火的蛋白质。 测定方法将 为不同类型的退火反应建立。 净化退火 蛋白质将根据它们与RNA的相互作用来表征 和促进碱基配对的能力。 单克隆抗体将是 针对退火蛋白制备并用于表征细胞内 定位和与RNA的关联作为理解 这些蛋白质的生物学作用。 这项工作对于 了解介导RNA-RNA碱基配对的蛋白质的性质， vivo. 因此，这项研究将为分子生物学提供进一步的见解。 调节高等生物基因表达的相互作用及其 病毒并增强我们在体内调节这些相互作用的能力， 体外

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.