MECHANISMS OF P53 MEDIATED APOPTOSIS AND CARCINOGENESIS

作为了解P53介导的机制的一步 细胞凋亡与癌变，我们发现了一种新的可能的锌 手指转录因子Pw1/asf-1和两个CED3/ICE相关基因 蛋白酶、NEDD2和CPP32，其基因表达被诱导 特别是在P53介导的细胞凋亡过程中。这样做的主要目标是 研究项目是研究这三个因素的作用和调节。 P53介导的细胞凋亡中的基因产物。在第一个具体目标中， Pw1/ASF-1在P53介导过程中的诱导意义 将对细胞凋亡进行检测。这种蛋白质的生物学功能将是 PW/ASF-1基因产物在哺乳动物中的表达分析 细胞。Pw1/ASF-1蛋白作为转录因子的功能 将在特定目标中进行探索2.鉴定DNA的实验 提出了一种体外免疫选择结合位点的方法。 Pw1/ASF-1基因表达的调控机制也将是 通过分析其启动子和增强子元件进行研究。在 第三个特定目标，调控CED3/ICE相关的蛋白酶，NEDD2 和CPP32，在p53介导的细胞凋亡过程中将被检测。这个 蛋白水解酶基因表达的诱导可以作为一种 将测试激活CED3/ICE相关酶的机制。 P53基因改变经常与人类癌症有关 而P53介导的细胞凋亡在预防 癌变和恶性肿瘤，从而了解其发病机制 P53诱导的细胞凋亡是一个重要的研究途径 可能为更好地治疗人类肿瘤提供依据。

As a step toward understanding the mechanisms of p53 mediated apoptosis and carcinogenesis, we have identified a new putative zinc finger transcription factor, Pw1/ASF-1, and two CED3/ICE-related proteases, NEDD2 and CPP32, whose gene expressions are induced specifically during p53 mediated apoptosis. The main goal of this research project is to investigate the role and regulation of these three gene products in p53-mediated apoptosis. In the first specific aim, significance of the induction of Pw1/ASF-1 during p53 mediated apoptosis will be examined. Biological functions of this protein will be analyzed by expressing the Pw/ASF-1 gene product in mammalian cells. How the Pw1/ASF-1 protein functions as a transcription factor will be explored in specific aim 2. Experiments to identify the DNA binding sites by an in vitro immuno-selection method are proposed. Regulatory mechanism of the Pw1/ASF-1 gene expression will also be investigated by analyzing its promoter and enhancer elements. In the third specific aim, regulation of CED3/ICE related proteases, NEDD2 and CPP32, during p53 mediated apoptosis will be examined. The hypothesis that Induction of protease gene expression can serve as a mechanism to activate CED3/ICE related proteases will be tested. Genetic alteration of p53 is frequently associated with human cancers and p53-mediated apoptosis plays a critical role to prevent carcinogenesis and malignancies, Thus, understanding the mechanism of p53-induced apoptosis represents an important research avenue and may provide a basis for better treatment of human tumors.