POLY-ADP-RIBOSE AND REPAIR OF RADIATION INDUCED DAMAGE

聚 ADP 核糖和辐射损伤的修复

基本信息

批准号：
3192532
负责人：
THOMAS Dominic STAMATO
金额：
$ 8.72万
依托单位：
LANKENAU INSTITUTE FOR MEDICAL RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
1990
资助国家：
美国
起止时间：
1990-09-25 至 1991-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3192532
关键词：
DNA repair DNA replication adenosine diphosphate cell cycle hamsters ionizing radiation ligase molecular genetics mutant nicotinamide adenine dinucleotide nucleic acid biosynthesis radiation genetics radiation sensitivity relative biological effectiveness

项目摘要

The polymer poly(ADP-ribose) is present is the nuclei of all eukaryotic organisms. While a number of studies have suggested the involvement of this polymer in the repair of damage produced by ionizing radiation and other agents, the biological role of this polymer in DNA repair still remains unclear. Thus, the ultimate goal of this research is to determine what the nature of this involvement might be in mammalian cells. To answer this question, we propose to isolate mutants in the Chinese hamster ovary cell which are deficient in the synthesis of poly(ADP-ribose) and determine the physiologic and biochemical consequences of this deficiency. These mutants will be isolated using a non-selective screening approach in which colonies are examined in situ for enzyme activity using radioactive substrate (NAD) and autoradiography. Viable cells from putative enzyme-deficient colonies will be recovered from nylon cloth replicas and tested for poly(ADP-ribose) enzyme activity in cell free extracts. Poly(ADP-ribose) polymerase negative mutants will be examined for abnormal sensitivity to killing by gamma-rays and other DNA damaging agents. Those agents which produce an increase in sensitivity will also be examined for sensitivity through the cell- cycle. The kinetics of repair of single-and double-strand DNA breaks will be examined for those agents which produced increased sensitivities. Also repair replication and unschedule DNA synthesis will be examined. We believe that biochemical characterization of a poly(ADP)-ribose-deficient mutant offers the prospect of elucidating the role of this polymer in DNA repair.

存在聚合物聚（ADP-核糖）是所有的核真核生物。虽然许多研究表明该聚合物参与修复由电离辐射和其他药物，这是生物学的作用 DNA修复中的聚合物仍然不清楚。因此，最终这项研究的目标是确定这一点的本质参与可能是在哺乳动物细胞中。要回答这个问题，我们建议在中国仓鼠卵巢细胞中分离突变体在综合poly（adp-ribose）和确定此的生理和生化后果不足。这些突变体将使用非选择性分离筛选方法，在原位检查菌落使用放射性底物（NAD）和放射自显影。假定酶的生存细胞殖民地将从尼龙布复制品中回收并测试用于无细胞提取物中的聚（ADP-核糖）酶活性。将检查聚（ADP-核糖）聚合酶阴性突变体的阴性突变体对被伽马射线和其他DNA杀死的异常敏感性有害代理。那些产生增加的代理商还将检查灵敏度通过细胞的灵敏度循环。单链DNA修复的动力学将为那些产生增加的代理人检查中断敏感性。还要修复复制和非安排DNA 将检查合成。我们相信生化 poly（ADP） - ribose缺陷突变体的表征提供阐明该聚合物在DNA修复中的作用的前景。