EIA CONTROL OF PROTEASE GENE TRANSCRIPTION

蛋白酶基因转录的 EIA 控制

• 批准号: 3468149
• 负责人: Steven Miles Frisch
• 金额: $ 12万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3468149
• 关键词: DNA footprinting; HeLa cells; collagenase; gene induction /repression; genetic regulatory element; genetic transcription; metastasis; microinjections; oncogenes; oncoproteins; transcription factor; transfection

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.

分析耦合细胞的机制和组件 信号转导，转录响应和细胞生长的过程 是该研究赠款的申请人的主要职业兴趣。 的 特别感兴趣的是癌症调节的机制 这些过程。 激活的癌基本质上是一个突变基因， 提供表型，揭示行为的不断新方面， 调节分子的设计和相互作用。 在这个框架中， 拟议的研究有望产生有关互动的新见解 具有两个不同调控基因的E1a癌基因的间隙 胶原酶（CL）和IV型胶原酶（T4）。 蛋白水解酶 这些基因编码在肿瘤转移中起重要作用。 E1a 蛋白质将用作分析其调节的关键探针。 具体而言，Cl基因启动子的增强子元素TPA- 该研究者发现调节元件是E1A调节的 增强剂：E1A在一个单元线中抑制其活性，但在将其激活 其他。 识别对E1A响应的因素或 通过从功能上测定一个细胞的蛋白质来实现负面影响 在另一个背景中的系统。 为此目的的测定将是 两种一般类型，体外（例如，体外转录）和体内 （转染或显微注射。）T4基因调节区域将是 进一步分析，因为已经发现增强剂和消音器元素。 一个结合AP2样转录的E1A可抑制增强子元件 因子将更详细地表征和E1a的机制 抑制作用将进行分析。