EIA CONTROL OF PROTEASE GENE TRANSCRIPTION

耦合细胞的机制和成分分析 信号转导、转录反应和细胞生长的过程 是这项研究补助金申请人的主要职业兴趣。 的 特别令人感兴趣的是癌基因蛋白调节 这些过程。 激活的癌基因本质上是一种突变基因， 提供了一种不断揭示行为新方面的表型， 调控分子的设计和相互作用。 在此框架内， 拟议中的研究有望产生关于相互作用的新见解， E1 A癌基因与两个不同的调控基因，间质 胶原酶（CL）和IV型胶原酶（T4）。 蛋白水解酶 这些基因编码的蛋白在肿瘤转移中起重要作用。 E1a 蛋白质将被用作分析其调控的关键探针。 具体地，CL基因启动子的增强子元件，TPA- 该研究者发现监管要素是E1 A监管的 增强子：E1 A在一种细胞系中抑制其活性，但在另一种细胞系中激活其活性。 另 确定对E1 A有积极反应的因素， 通过对来自一个细胞的蛋白质进行功能性分析， 系统在另一个的背景下。 用于此目的的测定将是 两种一般类型，在体外（例如，体外转录）和体内 （转染或显微注射。） T4基因调控区将是 进一步分析，因为已经发现了增强剂和沉默剂元素。 一个结合AP 2样转录的E1 A抑制性增强子元件 因子将被更详细地表征，并且E1 A的机制 将对压制进行分析。

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.