DNA Polymerase Beta and Cell Transformation

拟议研究的广泛的长期目标是确定异常的碱基切除修复 (BER)与癌症有关。该项目的具体目标是： 1.检验细胞转化活性是Pol~癌变相关细胞共同属性的假设 变种。我们将专注于我们尚未表征的变体，并完成 有趣的POL~肿瘤相关变异体的特征。我们将使用一种组合的遗传、生化 和细胞生物学方法来确定细胞转化的机制，这是由 变种。 2.验证肿瘤相关变异诱导小鼠肿瘤发生的假说。“敲进去”的老鼠会 使用标准方法构建。肿瘤的发生将作为年龄的函数进行监测，也将在 一种或多种DNA损伤剂的存在。 3.检验BER异常和病毒感染相结合导致 肿瘤发生学。在AIM 3中构建的小鼠将用于HPV的皮肤模型，以确定是否存在病毒 蛋白质与异常的BER协同作用以降低或潜伏期或提高肿瘤形成或 转移。其他各种在误码率方面有缺陷的小鼠也将在这一目标中表现出来。 4.验证microRNAs在BER调节中的重要作用这一假说。来检验这一假设 BER基因中microRNA靶点的改变会导致与癌症相关的异常BER。我们会 鉴定已知的生殖系中保守的微小RNA靶点的单核苷酸多态 特异性BER基因的3‘UTRs。我们还将确定人类肿瘤中的microRNA靶标是否发生了变化。

The broad long-term objective of the proposed research is to determine how aberrant base excision repair (BER) is linked to cancer. The Specific Aims of this project are: 1. To test the hypothesis that cellular transforming activity is a common property of the Pol ~ cancerassociated variants. We will focus on variants we have not yet characterized and complete the characterization of interesting Pol ~ tumor-associated variants. We will use a combined genetic, biochemical and cell biological approach to identify the mechanism of cellular transformation that is employed by the variants. 2. To test the hypothesis that cancer-associated variants induce tumorigenesis in mice. "Knock-in" mice will be constructed using standard methods. Tumorigenesis will be monitored as a function of age and also in the presence of one or more DNA damaging agents. 3. To test the hypothesis that a combination of aberrant BER and viral infection leads to tumorigenesis. The mice constructed in Aim 3 will be used in a skin model of HPV to determine if viral proteins synergize with aberrant BER to decrease the rate or latency or increase the rate of tumorigenesis or metastasis. Various other mice with deficiencies in BER will also be characterized in this aim. 4. To test the hypothesis that microRNAs are important in the regulation of BER. To test the hypothesis that alterations in microRNA target sites in BER genes results in aberrant BER that is linked to cancer. We will characterize known germ line single nucleotide polymorph isms in conserved micro RNA target sites within the 3'UTRs of specific BER genes. We will also determine if microRNA targets are altered in human tumors.