Characterization of Fortilin, A Novel Anti-p53 Protein

Fortilin（一种新型抗 p53 蛋白）的表征

• 批准号: 6365303
• 负责人: Ken Fujise
• 金额: $ 22.43万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6365303
• 关键词: Bax gene /protein; apoptosis; atherosclerotic plaque; cell growth regulation; inhibitor /antagonist; p53 gene /protein; protein binding; protein protein interaction; protein structure function

DESCRIPTION (provided by the applicant): Atherosclerosis is the leading cause of coronary artery disease, peripheral vascular disease and cerebrovascular disease in this country, afflicting millions of Americans. The pathogenesis of atherosclerosis is complex, yet dysregulated proliferation of smooth muscle cells is implicated as an important proponent. A genetic study using p53 knockout mice suggested that he presence of functional 053 is preventive against atherosclerosis in a hyperlipidemic environment. In addition, studies have indicated that the presence of p53 inhibitors, such as Mdm-2 and IE84, accelerates atherosclerosis. Our laboratory identified a new p53 inhibitor and designated it as fortilin. Characterization of fortilin in our laboratory so far showed: (1) The peptide sequence of fortilin is highly conserved among different species. (2) The message of fortilin is resent in all normal human tissues while its expression is greater in cancerous cell lines. (3) Fortiin expression is more prominent in malignant than I benign tissue. (4) Fortilin binds p53. (5) The overexpression prevents of fortilin prevents cells from undergoing etoposide-induced apoptosis. (6) The over expression of fortilin prevents cells from undergoing apoptosis induced by the expression of p53. (7) The overexpression of fortilin inhibits p53-mediated transactivation of BAX, a pro-apoptotic molecule. (8) The expression of fortilin is up regulated in atheroma. There are three major Specific Aims proposed to further characterize fortilin and the fortilin-p53 interaction: Aim 1: To define the region of fortilin that interacts with p53 and other molecules. We will first determine which domain of fortilin participates in p53 binding by generating and evaluating various deletion mutants for their ability to interact with fortilin (Aim1.1). We will then identify the region of that fortilin domain that is required for the interaction with p53 (Aim1.2). Finally, we will establish whether fortilin is covalently modified by sentrin and other ubiquitin-like molecules (Aim 1.3). Aim 2: To characterize the region of p53 that interacts with fortilin and other molecules. In this major aim, we determine which domain (Aim 2.1) and region (Aim 2.2) of p53 participates in fortilin binding, using deletion and point mutants, respectively. We will then evaluate whether the fortilin-binding region of p53 is required for p53 to interact with Mdm2, Mdm-X and BRCA2, p53 binding and inhibitory molecules (Aim 2.3). Aim 3: To investigate the regulatory role of fortilin-p53 interaction in apoptosis. We will determine whether fortilin inhibits p53-mediatd apoptosis (Aim3.1), p53-mediated transactivation of Bax (Aim 3.2) and p53 binding to the Bas-responsive element (Aim 3.3) THROUGH ITS DIRECT binding of p53. Knowledge gained I this proposal will enable us to have a solid foundation for the future intervention of human atherosclerosis.

描述（由申请人提供）：动脉粥样硬化是主要原因 冠状动脉疾病、周围血管疾病和脑血管疾病 这个国家的疾病困扰着数百万美国人。发病机制 动脉粥样硬化很复杂，但平滑肌增殖失调 细胞被认为是一个重要的支持者。使用 p53 的遗传学研究 基因敲除小鼠表明功能性 053 的存在具有预防作用 在高脂血症环境下对抗动脉粥样硬化。此外，研究 已经表明 p53 抑制剂的存在，例如 Mdm-2 和 IE84， 加速动脉粥样硬化。我们的实验室发现了一种新的 p53 抑制剂 将其命名为福替林。我们实验室对福替林的表征 远期表明：（1）fortilin的肽序列在不同物种之间高度保守。 不同的物种。 (2) 福替林的信息在所有正常人体内均不存在 组织中，而其在癌细胞系中表达更高。 (3) 福廷 表达在恶性组织中比在良性组织中更突出。 (4) 福替林 结合 p53。 (5) fortilin的过表达阻止细胞 经历依托泊苷诱导的细胞凋亡。 (6) fortilin过度表达 防止细胞发生由 p53 表达诱导的细胞凋亡。 (7) Fortilin 的过表达抑制 p53 介导的 BAX 反式激活，BAX 是一种 促凋亡分子。 (8) fortilin表达上调 动脉粥样硬化。提出了三个主要具体目标来进一步描述 fortilin 和 fortilin-p53 相互作用：目标 1：定义 fortilin 与 p53 和其他分子相互作用。我们首先会确定 fortilin 的哪个结构域通过生成和参与 p53 结合 评估各种缺失突变体与 fortilin 相互作用的能力 （目标1.1）。然后我们将确定 fortilin 结构域的区域 与 p53 相互作用所需的 (Aim1.2)。最后，我们将建立 fortilin是否被sentrin和其他泛素样蛋白共价修饰 分子（目标 1.3）。目标 2：表征 p53 相互作用的区域 与福替林和其他分子。在这个主要目标中，我们确定哪个域 p53 的（目标 2.1）和区域（目标 2.2）参与 fortilin 结合，使用 分别是缺失突变体和点突变体。然后我们将评估是否 p53 需要 p53 的 fortilin 结合区才能与 Mdm2、Mdm-X 相互作用 以及 BRCA2、p53 结合和抑制分子（目标 2.3）。目标 3： 研究 fortilin-p53 相互作用在细胞凋亡中的调节作用。我们 将确定 fortilin 是否抑制 p53 介导的细胞凋亡 (Aim3.1)， p53 介导的 Bax 反式激活（目标 3.2）和 p53 与 碱性响应元件（目标 3.3）通过直接结合 p53。知识 我认为这个建议将使我们为未来打下坚实的基础 干预人体动脉粥样硬化。