CELL CYCLE REGULATION OF DNA REPAIR

拟议研究的总体目标(S)是为了获得一个基本的 对时间调控机制的理解(S) DNA修复基因的表达。彻底分析监管机构 参与调节DNA修复过程的机制将会增加 我们对真核生物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永续的策略的了解 没有错误。从理解规则中获得的信息 DNA修复过程的表达应该有助于深入了解 DNA修复与遗传性疾病的关系 易患癌症是一种表型特征。 DNA修复过程的时间调制似乎是一种常见的 在不同来源的哺乳动物细胞中具有这种特征。此外，还有 似乎与DNA细胞周期控制的改变之间存在关联 修复过程和DNA修复缺陷或对损伤过敏。 然而，这种调节的确切机制尚不清楚。几个 问题仍然没有得到回答：调制是在 转录还是转录后？控制是否在翻译时启动 或翻译后修改；或者，它可以是 所有这些都结合在一起。但很明显，人类和 CHO细胞将为理解调控机制提供良好的模型 涉及的流程。 作为理解监管机制的持续努力的一部分(S) 参与DNA修复过程的时间调制以及 Bloom综合征遗传缺陷及其修复的分子基础 缺乏CHO细胞系，建议检查DNA的控制 在分子水平上的修复过程。它计划着眼于控制 机制从几个不同的方面；1)检查是否 控制处于转录水平。计划对信使核糖核酸进行定量 用Northern研究细胞周期中各种DNA修复基因的水平 斑点杂交；2)克隆和测序 中国仓鼠细胞中的脱嘌呤/脱嘧啶(AP)内切酶。它是 计划确定是否存在DNA序列异常 正常和修复缺陷的AP内切酶可以解释 观察到时间调制的差异；3)构建基因组 来自CHO细胞的文库。将对生成的文库进行尿嘧啶的筛选 DNA糖基酶(UDG)和AP内切酶基因寻找共同点 可能解释这些基因协调控制的调控区 产品；4)学生培训是拟议的 研究。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 科学努力的严酷。

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.