EIA CONTROL OF PROTEASE GENE TRANSCRIPTION

耦合细胞的机理和成分分析 信号转导、转录反应和细胞生长过程 是这项研究资助申请者的主要职业兴趣。的 特别令人感兴趣的是癌基因蛋白调节的机制。 这些过程。被激活的癌基因本质上是一个突变基因，即 提供了一种不断揭示行为新方面的表型， 调节分子的设计和相互作用。在这个框架中， 拟议中的研究有望对相互作用产生新的见解 具有两个不同调控基因的E1a癌基因 胶原酶(CL)和IV型胶原酶(T4)。蛋白水解酶 由这些基因编码的基因在肿瘤转移中起着重要作用。E1a 蛋白质将被用作关键探针来分析它们的调控。 具体地说，CL基因启动子的增强子元件TPA- 调节元件，被这位调查者发现是E1a调节的 增强子：E1a在一个细胞系中抑制其活性，但在 又一个。确定对E1a有积极或积极反应的因素 通过对一个细胞中的蛋白质进行功能分析，将实现阴性 系统在后台的另一个。为此目的进行的化验将包括 两种一般类型，体外(例如，体外转录)和体内 (转染法或显微注射法)T4基因调控区将是 进一步分析，因为已经发现了增强器和消音器元件。 一个E1a可阻抑的增强子元件，可结合AP2样转录 我们将更详细地描述E1a的作用机制 压制将被分析。

Analysis of the mechanisms and components underlying the coupled cellular processes of signal transduction, transcriptional response and cell growth is the major career interest of the applicant for this research grant. Of particular interest is the mechanism by which oncogene proteins regulate these processes. The activated oncogene is essentially a mutant gene, that provides a phenotype revealing continually new aspects of the behavior, design and interactions of regulatory molecules. In this framework, the proposed research promises to yield new insights concerning the interaction of the E1A oncogene with two dissimilarly regulated genes, interstitial collagenase (CL) and type IV collagenase (T4). The proteolytic enzymes encoded by these genes play an important role in tumor metastasis. E1A protein will be used as a critical probe to analyze their regulation. Specifically, an enhancer element of the CL gene promoter, the TPA- Regulatory Element, was found by this investigator to be an E1A-regulated enhancer: E1A represses its activity in one cell line, but activates it in another. The identification of factors that respond to E1A positively or negatively will be achieved by functionally assaying proteins from one cell system in the background of the other. Assays for this purpose will be of two general types, in vitro (e.g., in vitro transcription) and in vivo (transfection or microinjection.) The T4 gene regulatory region will be analyzed further, as both enhancer and silencer elements have been found. One E1A-repressible enhancer element that binds an AP2-like transcription factor will be characterized in more detail, and the mechanism of E1A repression will be analyzed.