Genome-wide repair maps of nucleotide excision repair in Drosophila and S2 cells.

We report the application of single-molecule-based sequencing technology (XR-seq) for high-throughput profiling of nucleotide excision repair in Drosophila four developmental stages and S2 cells. By obtaining over six billion bases of sequence from UV and cisplatin damage antibodies immunoprecipitated excision DNA, we generated genome-wide nucleotide excision repair maps of Drosophila Embryo, Larva, Pupa , two gender of adults and S2 cells . We find that Drosophila performs transcription-coupled repair (TCR) at all its developmental stages. S2 cell carry out TCR response to both UV and Cisplatin damage. Finally, we show that XPC repair factor is required for both global and transcription-coupled repair in Drosophila. This study provides the mechanism of nucleotide excision repair of Drosophila in vivo and vitro response to UV and Cisplatin damage. Overall design: Examination of TCR in all Drosophila whole life cycle and gender abd the function of XPC repair factor.

本研究报道了基于单分子的测序技术（XR-seq）在果蝇四个发育阶段及S2细胞的核苷酸切除修复（nucleotide excision repair）高通量谱型分析中的应用。通过从经靶向紫外线（UV）和顺铂（cisplatin）损伤的抗体免疫沉淀的切除DNA中获取超过60亿碱基的序列数据，我们构建了果蝇胚胎、幼虫、蛹、雌雄成虫及S2细胞的全基因组核苷酸切除修复图谱。研究发现，果蝇在所有发育阶段均执行转录偶联修复（transcription-coupled repair, TCR）；S2细胞可对紫外线及顺铂损伤产生转录偶联修复应答。最终，我们证实果蝇的全局修复与转录偶联修复均依赖XPC修复因子。本研究阐明了果蝇体内外针对紫外线及顺铂损伤的核苷酸切除修复机制。整体实验设计：对果蝇全生命周期及不同性别中的转录偶联修复进行检测，并探究XPC修复因子的功能。