RNA-RNA ANNEALING PROTEINS

RNA-RNA 退火蛋白

• 批准号: 3438897
• 负责人: STEPHEN H MUNROE
• 金额: $ 9.97万
• 依托单位: MARQUETTE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-04-01 至 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3438897
• 关键词: HeLa cells; RNA; RNA splicing; antisense nucleic acid; conformation; double stranded RNA; gel electrophoresis; heterogeneous nuclear ribonucleoprotein; immunofluorescence technique; monoclonal antibody; nucleic acid hybridization; nucleobase; protein purification; protein structure function; ribonucleoproteins

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.

本研究的目的是分离和表征HeLa蛋白

项目成果

Inhibition of c-erbA mRNA splicing by a naturally occurring antisense RNA.

• DOI: 10.1016/s0021-9258(18)54535-x
• 发表时间: 1991-11
• 期刊: The Journal of biological chemistry
• 作者: S. H. Munroe;Mitchell A. Lazar