CYTOKINE GENE REGULATION--CONTROL OF MRNA STABILITY

该项目的长期目标是确定 来调节信使RNA的稳定性。尽管我们已经学到了很多东西 关于转录基因调控，我们知之甚少 控制信使RNA稳定性的因素。它越来越多地 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mRNAs稳定或 对外界刺激的反应被降解的情况还没有确定。在……里面 这个项目，人类转录后调控的机制 炎性细胞因子基因，由白介素1(IL-1) 调查过了。满足以下条件的最小序列 IL-1对信使核糖核酸稳定性的依赖调节将由 改变的gro基因的转染法。的功能意义 预测的调控序列中的二级结构元件将 通过定点突变进行检测。细胞质因子 将使用In来表征与调控序列的相互作用 体外方法。我们将研究gro基因在体内的衰退过程。 确定核酸酶攻击的初始位置并识别 控制gro信使核糖核酸周转的药理作用。一种体外系统 将被开发用来分离调节gro基因的因子。 稳定性。这个项目将增加对分子事件的理解 控制炎症反应，并应该阐明一种机制 以生物反应为基础的基因调控。

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.