FUNCTIONAL ANALYSIS OF ETS1 PROTEIN ISOTYPES

ETS1 蛋白同种型的功能分析

• 批准号: 2763895
• 负责人: TAKIS S PAPAS
• 金额: $ 12.99万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2763895
• 关键词: DNA binding protein; apoptosis; carcinogenesis; cell growth regulation; epithelium; gel mobility shift assay; gene expression; genetic library; genetic regulation; immunoprecipitation; neoplastic cell; nucleic acid sequence; oncogenes; phenotype; phosphorylation; polymerase chain reaction; protein isoforms; tetracyclines; tissue /cell culture; transcription factor; transfection

Cancer is a multi-step disease involving a series of genetic alterations that result in dysregulation of cell proliferation and differentiation. These sporadic mutations, some of which are caused by factors such as environmental chemicals and radiation, lead to the activation of oncogenes, the loss of tumor suppression and the activation of programmed cell death. Our laboratory has been involved for many years in studies dealing with the molecular biology of the ets gene family members. Having been the first group to demonstrate that these genes exist as a family, we cloned and characterized the first several family members, including ETS1, ETS2, ERG, ERGB, and ERF. While it has been shown that ETS1 is an oncogene when it is transduced by the E26 virus, we have determined the role of many of ets genes in cell proliferation, normal differentiation, and in some cases abnormal development leading to cancer. The experiments described in this proposal are designed to understand in detail the mechanism by which an isotype of ETS1, generated as a splicing variant and referred to as p42-ETS1, represses tumorigenicity and induces apoptosis in epithelial cells. To accomplish these goals we will: i) analyze the molecular mechanism by which the p42-ETS1 protein leads to the induction of apoptosis in epithelial cancer cells; ii) determine which are the critical DNA regions (domains) within the p42-ETS1 gene that are required for the ability of the gene product to induce apoptosis in epithelial cancer cells; and iii) identify and characterize the function of p42-ETS1 target genes involved in the induction of apoptosis in epithelial cancer cells, using a tetracycline inducible system. This work can have profound medical significance in that it may open new insights as to the potential role of the p42-ETS1 splice variant ETS1 isotype in controlling cell proliferation by the induction of apoptosis in epithelial cell cancers. Such insights may provide a basis for the design of a gene therapy approach utilizing the ability of p42-ETS1 to initiate cell death in cancer cells.

癌症是一种多步骤疾病，涉及一系列导致细胞增殖和分化失调的遗传改变。 这些零星突变，其中一些是由环境化学品和辐射等因素引起的，导致癌基因的激活，肿瘤抑制的丧失和程序性细胞死亡的激活。 我们的实验室多年来一直从事ets基因家族成员的分子生物学研究。作为第一个证明这些基因作为一个家族存在的小组，我们克隆并表征了前几个家族成员，包括ETS 1，ETS 2，ERG，ERGB和ERF。 虽然已经表明，当它被E26病毒转导时，ETS 1是一种致癌基因，但我们已经确定了许多ets基因在细胞增殖、正常分化以及在某些情况下导致癌症的异常发育中的作用。本提案中描述的实验旨在详细了解ETS 1的同种型（作为剪接变体产生，称为p42-ETS 1）抑制致瘤性并诱导上皮细胞凋亡的机制。 为了实现这些目标，我们将：i）分析p42-ETS 1蛋白诱导上皮癌细胞凋亡的分子机制; ii）确定p42-ETS 1基因内哪些是基因产物诱导上皮癌细胞凋亡的能力所需的关键DNA区域（结构域）;和iii）使用四环素诱导系统鉴定和表征参与上皮癌细胞中细胞凋亡诱导的p42-ETS 1靶基因的功能。这项工作可能具有深远的医学意义，因为它可能会打开新的见解，p42-ETS 1剪接变异体ETS 1同种型在控制上皮细胞癌细胞凋亡诱导细胞增殖的潜在作用。 这些见解可以为利用p42-ETS 1启动癌细胞死亡的能力设计基因治疗方法提供基础。