FUNCTIONAL ANALYSIS OF ETS1 PROTEIN ISOTYPES

ETS1 蛋白同种型的功能分析

• 批准号: 6203462
• 负责人: TAKIS S PAPAS
• 金额: $ 15.41万
• 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: 1999-08-25 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6203462
• 关键词: T lymphocyte; apoptosis; carcinogenesis; cell growth regulation; cell line; developmental immunology; gene expression; gene targeting; genetic regulation; genetically modified animals; laboratory mouse; molecular site; mutant; neoplasm /cancer genetics; nucleic acid sequence; phenotype; phosphorylation; protein isoforms; tetracyclines; transcription factor

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/Fli1, 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. The potentially important role of the p42- ETS1 protein in the maintenance of normal T-cell populations will also be examined. 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 in T-cells, 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, and iv) study the normal function of the p42-ETS1 in T-cells by generating knock-out mice individually lacking one of the ETS1 isotypes. 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.

癌症是一种涉及一系列基因改变的多步骤疾病 导致细胞增殖和分化失调。 这些偶发突变，其中一些是由以下因素引起的， 环境化学品和辐射，导致激活 癌基因，肿瘤抑制的丧失和程序性癌基因的激活， 细胞死亡我们的实验室多年来一直致力于 处理ets基因家族成员的分子生物学。具有 作为第一个证明这些基因作为一个家族存在的小组，我们 克隆并鉴定了前几个家族成员，包括ETS 1， ETS 2、ERG、ERGB/Fli 1和ERF。虽然已经表明ETS 1是一种 当它被E26病毒转导时，我们已经确定了 许多ETS基因在细胞增殖，正常分化， 并且在某些情况下导致癌症的异常发育。 本提案中描述的实验旨在了解 详细介绍了机制，其中ETS 1的同种型，作为剪接产生 一种称为p42-ETS 1的变体，抑制肿瘤发生并诱导 上皮细胞凋亡。p42的潜在重要作用- ETS 1蛋白在维持正常T细胞群中的作用也将是 考察为了实现这些目标，我们将，i）分析分子 p42-ETS 1蛋白导致诱导 上皮癌细胞凋亡，ii）确定哪些是关键的 p42-ETS 1基因内的DNA区域（结构域），这些区域是 基因产物诱导上皮癌细胞凋亡的能力 和在T细胞中，iii）鉴定和表征p42-ETS 1的功能， 上皮癌细胞凋亡诱导中的靶基因 细胞，使用四环素诱导系统，和iv）研究正常的 通过产生基因敲除小鼠在T细胞中p42-ETS 1的功能 单独缺乏ETS 1同种型之一。 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 关于p42-ETS 1剪接变体ETS 1的潜在作用的见解 同种型通过诱导细胞凋亡控制细胞增殖， 上皮细胞癌这样的见解可以为设计提供基础 利用p42-ETS 1启动基因治疗的能力， 癌细胞的死亡。