Regulation of PEP-carboxykinase mRNA Breakdown by cAMP

cAMP 对 PEP-羧激酶 mRNA 分解的调节

• 批准号: 9106256
• 负责人: Yaacov Hod
• 金额: $ 19.3万
• 依托单位: SUNY at Stony Brook
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1995-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9106256&HistoricalAwards=false
• 关键词: Regulation; PEP; carboxykinase; mRNA; Breakdown

The steady-state level of specific mRNA is determined by the rates by which this molecule is synthesized and degraded. Despite the importance of mRNA turn-over in the control of gene expression, the mechanisms that underlie this process are still obscure. It is thus the long-term goal of the proposed research to explore the mechanism of intracellular mRNA breakdown. In recent studies the PI has shown that the breakdown rate of the mRNA for PEP- carboxykinase (PEPCK) is hormonally regulated and that cAMP (as an intracellular mediator of glucagon) stabilizes the mRNA against degradation. It has also been demonstrated that 733 nucleotides from the 3'-region of the mRNA are sufficient to confer the cAMP regulation of mRNA turn-over. The main goal of this proposal is to delineate the nucleotide domains that mediate the cAMP regulation of PEPCK mRNA as well as the determinants that confer the short half-life of PEPCK mRNA. These studies will involve monitoring the half-lives of mRNA transcripts of a variety of hybrid genes containing different sequences of PEPCK, after these genes have been introduced into hepatoma cells. In addition, we propose to examine the role of RNA-protein interaction at the cAMP- responsive element in the control of the half-life of PEPCK mRNA. It is hoped that these studies will result in a better understanding of the molecular events which control gene expression and also will find a practical use in the construction of an efficient expression system in which the control of mRNA stability is considered when a foreign gene is to be introduced into cells.//

特定mRNA的稳态水平是由该分子合成和降解的速率决定的。尽管mRNA翻转在基因表达控制中的重要性，但这一过程背后的机制仍然不清楚。因此，探索细胞内mRNA分解的机制是本研究的长期目标。在最近的研究中，PI表明PEP-羧激酶（PEPCK） mRNA的分解率是受激素调节的，cAMP（作为细胞内胰高血糖素的介质）稳定mRNA的降解。研究还表明，来自mRNA 3'区的733个核苷酸足以赋予cAMP对mRNA翻转的调节。本提案的主要目标是描述介导cAMP调控PEPCK mRNA的核苷酸结构域，以及赋予PEPCK mRNA短半衰期的决定因素。这些研究将包括监测多种含有不同PEPCK序列的杂交基因的mRNA转录物在将这些基因引入肝癌细胞后的半衰期。此外，我们建议研究cAMP响应元件上的rna -蛋白相互作用在控制PEPCK mRNA半衰期中的作用。希望这些研究将有助于更好地了解控制基因表达的分子事件，并在构建有效的表达系统中找到实际应用，在将外源基因引入细胞时考虑对mRNA稳定性的控制。//