Study on Genetic Recombination Systems in Eukaryotes

真核生物基因重组系统的研究

基本信息

批准号：
06101003
负责人：
OGAWA Hideyuki
金额：
$ 170.88万
依托单位：
Iwate College of Nursing (1998)Osaka University (1994-1997)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Specially Promoted Research
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06101003/
关键词：
non-homologous end joining nuclease activity of Mre11 protein RecA protein strand-transfer activity of Rad51 Rad52 protein REP protein initiation of meiotic recombination processing from DSB ends MRE11,RAD50&XRS2遺伝子 MRE11,RAD50 & XRS2遺伝子

项目摘要

We set three main subjects as follows. (1) Elucidation of away of recognition in abase sequence between two homologous DNA molecules. (2) Analysis of structures and functions of recombination apparatuses for initiation and formation of recombination intermediates. (3) Analysis of chromosome structures required for recombination process.The following are the prominent results that we obtained for a five-year period of this investigation. (1) Structure of RecA-DNA complexes was analyzed by NMR spectroscopy. Analysis revealed that the DNA structure in the complexes contained novel deoxyribose-base stacking and bases of the single-stranded DNA were spaced out nearly 0.5 run. This novel structure prompted us to propose a new model for a recognition mechanism of homologous base sequence in homologous pairing. (2) We analyzed the function of the Mre 11 protein that played a central role in the initiation of meiotic recombination. The Mre 11 protein was involved in DNA double-strand break (DSB … More ) formation and carried out processing from the DSBs with its nuclease activities. In addition, Mre 11 protein also carried out non-homologous end joining reaction of DSBs and was involved in illegitimate recombination through the reaction. These findings suggest that the Mre 11 protein acts at the junction of homologous recombination pathway and illegitimate recombination pathway and may provide a breakthrough in improving a low frequency of gene targeting in eukaryotic cells. (3) Strand-transfer activity was observed for Rad5l protein at a high level as RecA protein by addition of Rad52 and REP proteins in the reaction mixture. This finding will accelerate the analysis of molecular mechanism of formation of recombination intermediates in eukaryotes.All results obtained contributed directly to the research field with repair of DSBs produced by irradiation of ionizing radiation. Furthermore they proposed new research problems, what a relationship is existed between meiotic recombination and DNA damage check point system, and how the Mre 11 protein is invoked in maintenance of a telomere length or repeated nucleotide sequences. Less

我们将三个主要主题设置为如下。（1）在两个同源DNA分子之间阐明ABase序列中识别的识别。（2）重组和形成重组中间体的结构和功能的分析。（3）重组过程所需的染色体结构分析。以下是我们在这项投资的五年期间获得的突出结果。（1）通过NMR光谱法分析了RECA-DNA复合物的结构。分析表明，复合物中的DNA结构包含新型的脱氧核糖核酸碱基堆叠和单链DNA的碱基的碱基，几乎将其间隔为0.5运行。这种新颖的结构促使我们提出了一个新模型，以实现同源配对中同源碱基序列的识别机制。（2）我们分析了MRE 11蛋白的功能，该蛋白在减数分裂重组中起着核心作用。此外，MRE 11蛋白还进行了MRE 11蛋白，还进行了非同理末端连接DSB的反应，并通过反应参与了非法重组。这些发现表明，MRE 11蛋白在同源重组途径和非法重组途径的连接处起作用，并且可能在改善真核生物中基因靶向的低频率方面具有突破性。（3）通过在反应混合物中添加RAD52和REP蛋白，可以在高水平作为RECA蛋白的RAD5L蛋白中观察到链转移活性。这一发现将加速真核生物中重组中间体形成的分子机制。通过修复电离辐射的辐射，通过修复DSB，获得的所有结果都直接贡献了研究领域。此外，他们提出了新的研究问题，减数分裂重组和DNA损伤检查点系统之间存在什么关系，以及如何在维持端粒长度或重复的核苷酸序列中调用MRE 11蛋白。较少的