CYTOKINE GENE REGULATION--CONTROL OF MRNA STABILITY

该项目的长期目标是确定机制 调节信使RNA的稳定性。 尽管我们已经了解到 关于转录基因的调控，我们对转录基因的调控知之甚少。 控制信使RNA稳定性的因素。 越来越 认识到许多细胞因子基因是由mRNA的变化来调节的， 稳定性，但这些mRNA稳定或 对外界刺激的反应尚未确定。 在 这个项目，人类转录后调控的机制， 炎症细胞因子基因，gro，通过白细胞介素-1（IL-1）， 研究了 必要且充分的最小序列， 赋予mRNA稳定性的IL-1依赖性调节将通过 改变的GRO基因的转染。 的功能意义 调控序列中预测的二级结构元件将 通过定点诱变进行测试。 细胞质因子， 与调控序列相互作用的特征将使用一种 体外方法。 将检查体内gro mRNA衰减的过程 以确定核酸酶攻击的起始位点， 控制gro mRNA周转的药物。 体外系统 将开发能够分离调节gro mRNA的因子 稳定 该项目将增加对分子事件的理解 控制炎症反应并阐明一种机制 对生物反应至关重要的基因调控。

The long-term objective of this project is to identify the mechanisms that regulate messenger RNA stability. Although much has been learned about transcriptional gene regulation, very little is known about the factors that control messenger RNA stability. It is increasingly recognized that many cytokine genes are regulated by changes in mRNA stability, but the mechanisms by which these mRNAs are stabilized or degraded in response to external stimuli have not been identified. In this project, the mechanisms of posttranscriptional regulation of a human inflammatory cytokine gene, gro, by interleukin-1 (IL-1) will be investigated. The minimum sequences that are necessary and sufficient to confer IL-1-dependent regulation of mRNA stability will be determined by transfection of altered gro genes. The functional significance of predicted secondary structural elements in the regulatory sequences will be tested by site-directed mutagenesis. Cytoplasmic factors that interact with the regulatory sequences will be characterized using an in vitro approach. The process of gro mRNA decay in vivo will be examined to determine the initial sites of nuclease attack and to identify pharmacologic agents that control gro mRNA turnover. An in vitro system will be developed to enable isolation of factors that regulate gro mRNA stability. This project will increase understanding of molecular events that control the inflammatory response and should elucidate a mechanism of gene regulation fundamental to biologic responses.