FUNCTION OF SNRNP PARTICLES IN PRE-MRNA SPLICING

SNRNP 颗粒在 Pre-mRNA 剪接中的功能

• 批准号: 3467690
• 负责人: Brian C. Rymond
• 金额: $ 10.59万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3467690
• 关键词: DNA footprinting; RNA splicing; Saccharomyces; fungal genetics; gene expression; precursor mRNA; small nuclear RNA; spliceosomes; tissue /cell culture

The removal of introns from pre-messenger RNA (pre-mRNA) is a requirement for the synthesis of functional messenger RNA (mRNA) from many eukaryotic genes. Failure of the pre-mRNA splicing process can prevent gene expression and lead to disastrous consequences for the organism. Several human disorders, including a number of thalassaemias, result from splicing defects. In addition, pre-mRNA splicing is a regulated process which controls the normal expression patterns of numerous genes. The splicing reaction takes place on a large ribonucleoprotein (RNP) particle called the spliceosome. Within the spliceosome are a number of small nuclear RNAs (snRNAs) which, together with associated proteins, comprise the small nuclear RNP (snRNP) subunits of the spliceosome. The long term objective of this study is to determine the contribution of snRNPs to the rate and fidelity of pre-mRNA splicing. Specific attention will be placed on defining the role of the U1 snRNP particle in 5' splice site utilization. U1 snRNA contains a sequence of its 5' end which base pairs with the 5' splice site sequence of pre-mRNA molecules. The U1-specific, 70 kd protein binds U1 snRNA adjacent to this pre-mRNA binding domain. An immediate goal of this work will be to determine if the 70 kd protein influences 5' splice site recognition, cleavage, or ligation. The gene encoding the 70 kd protein will be isolated from Saccharomyces cerevisiae (yeast) and in vivo and in vitro assays will be developed to monitor 70 kd protein function. Genetic and biochemical experiments will be performed to characterize the RNA binding properties of the 70 kd protein and to determine the contribution of the U1 snRNA/70 kd protein complex to spliceosome formation and splicing.

从前信使RNA（pre-mRNA）中去除内含子是一种 合成功能性信使RNA（mRNA）的需要， 许多真核基因。 前体mRNA剪接过程的失败可以 阻止基因表达，并导致灾难性的后果， 有机体 几种人类疾病，包括一些地中海贫血症， 这是由于拼接缺陷造成的。 此外，前体mRNA剪接是一种 调节过程，控制正常表达模式， 许多基因。 剪接反应发生在一个大的核糖核蛋白（RNP） 一种叫做剪接体的粒子。 在剪接体中有许多 小核RNA（snRNA），与相关蛋白质一起， 包括剪接体的小核RNP（snRNP）亚基。 的 本研究的长期目标是确定 snRNP对前体mRNA剪接的速率和保真度的影响。 具体 我们将把注意力放在确定U1 snRNP粒子的作用上 在5 ′剪接位点利用方面。 U1 snRNA包含其5'端序列，该序列与5'端碱基配对 前mRNA分子剪接位点序列。 U1特异性，70 kd 蛋白结合邻近该前mRNA结合结构域的U1 snRNA。 一个 这项工作的直接目标是确定70 kd蛋白质是否 影响5'剪接位点识别、切割或连接。 基因 编码70 kd蛋白的酵母菌将被分离 酿酒酵母（酵母）和体内和体外试验将被开发， 监测70 kd蛋白功能。 遗传和生物化学实验 将进行表征70种蛋白的RNA结合特性 kd蛋白质，并确定U1 snRNA/70 kd的贡献 蛋白质复合物对剪接体形成和剪接的影响。