CELL CYCLE REGULATION OF DNA REPAIR

拟议研究的总体目标是获得基本的 理解涉及时间调节的机制， DNA修复基因的表达。 对监管的全面分析 参与DNA修复过程调节的机制将增加 我们对真核生物中细胞周期介导的基因调控的理解， 我们对细胞用来确保自我延续的策略的了解 没有错误。 通过了解《生物多样性公约》 DNA修复过程的表达应该提供了对 DNA修复和遗传疾病之间的关系， 癌症易感性是一种表型特征。 DNA修复过程的时间调节似乎是一种常见的 不同来源的哺乳动物细胞中的特征。 此外还有 DNA的细胞周期控制改变似乎与 修复过程和DNA修复缺陷或对损伤超敏反应。 然而，这种调节的确切机制尚不清楚。 几 问题仍然没有答案：调制是否处于 转录还是转录后？ 是否在转换时启动控制 或翻译后修饰;或者，它可以是 以上所有的组合。 很明显，无论是人类还是 CHO细胞将为理解调控机制提供良好的模型。 涉及的过程。 作为理解监管机制的持续努力的一部分 参与DNA修复过程的时间调节以及 Bloom综合征遗传缺陷的分子基础及其修复 缺陷的CHO细胞系，建议检查DNA的控制 修复过程在分子水平上。 它计划看控制 从几个不同方面的机制; 1）检查是否 控制在转录水平。 计划定量mRNA 使用北方DNA修复技术， 2）克隆和测序主要目的基因 来自中国仓鼠细胞的脱嘌呤/脱嘧啶（AP）核酸内切酶。 是 计划确定是否有任何DNA序列的不规则性之间 正常和修复缺陷型AP核酸内切酶可以解释 观察到的时间调制差异; 3）构建基因组 来自CHO细胞的文库。 将对所得文库进行尿嘧啶筛选 DNA糖基化酶（UDG）和AP核酸内切酶基因，以寻找共同的 调节区域，这可能解释这些基因的协调控制 产品; 4）学生培训是建议的一个组成部分 research.研究生将有机会接受培训， 现代分子生物学技术，同时发展对 科学努力的严格性。

The overall objective(s) of the proposed research is to gain a fundamental understanding of the mechanism(s) involved in the temporal regulation of the expression of DNA repair genes. A thorough analysis of the regulatory mechanisms involved in the regulation of DNA repair processes will increase our understanding of cell cycle mediated gene regulation in eukaryotes and our knowledge of the strategies cell use to ensure self perpetuation without error. The information gained on understanding the regulation of the expression of DNA repair processes should provide insight into the relationship between DNA repair and genetic diseases in which predisposition to cancer is a phenotypic characteristic. The temporal modulation of DNA repair processes appears to be a common feature in mammalian cells of different origins. In addition, there appears to be a correlation between altered cell cycle control of DNA repair processes and DNA repair deficiency or hypersensitivity to damage. However, the exact mechanism of this modulation is unclear. Several questions remain unanswered: Is the modulation at the level of transcription or post transcriptional? Is control initiated at translation or post translational modification; Alternatively, it could be a combination of all of the above. It is clear though, that both human and CHO cells will provide good models for understanding the regulatory processes involved. As part of a continuing effort to understand the regulatory mechanism(s) involved in the temporal modulation of DNA repair processes as well as the molecular basis for the genetic defect in Bloom's syndrome and repair deficient CHO cell lines, it is proposed to examine the control of DNA repair processes at the molecular level. It is planned to look at control mechanisms from several different aspects; 1) To examine whether or not control is at the level of transcription. It is planned to quantitate mRNA levels of various DNA repair genes throughout the cell cycle using Northern Blot hybridization; 2) To clone and sequence the major apurinic/apyrimidinic (AP) endonuclease from Chinese hamster cells. It is planned to determine if there are any DNA sequence irregularities between the normal and the repair deficient AP endonuclease which could explain the observed difference in temporal modulation; 3) To construct a genomic library from CHO cells. The resulting library will be screened for uracil DNA glycosylase (UDG) and AP endonuclease genes to look for common regulatory regions which may explain the coordinate control of these gene products; 4) Student training is an integral component of the proposed research. Graduate students will be given the opportunity to be trained in modern molecular biology techniques while developing an appreciation for the rigors of scientific endeavor.