Double-strand break repair assays determine pathway choice and structure of gene conversion events in Drosophila melanogaster.
Double-strand break repair assays determine pathway choice and structure of gene conversion events in Drosophila melanogaster.
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DOI：
10.1534/g3.113.010074
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发表时间：
2014-03-20
期刊：
影响因子：
--
通讯作者：
LaRocque JR
中科院分区：
文献类型：
--
作者：
Do AT;Brooks JT;Le Neveu MK;LaRocque JR
双链断裂 (DSB) 必须准确有效地修复，以维持基因组完整性。根据发生断裂的生物体、DSB 的基因组位置以及发生断裂的细胞周期阶段，DSB 可以通过同源重组 (HR)、非同源末端连接 (NHEJ) 或单链退火 (SSA) 进行修复。开发了两种新型 DSB 修复测定法来确定这些修复途径的贡献并精细解析果蝇中的修复事件结构。 Rad51依赖性同源重组是有丝分裂细胞中首选的DSB修复途径，HR和SSA之间的途径选择发生在末端切除之后和Rad51依赖性链侵入之前。 HR 事件与长基因转换束相关，并且是双向和单向的，与通过合成依赖的链退火途径进行修复一致。此外，果蝇中不同序列之间的 HR 受到抑制，与人类细胞中报道的水平相似。对罕见 NHEJ 事件的连接分析表明，该系统使用了规范 NHEJ。
Double-strand breaks (DSBs) must be accurately and efficiently repaired to maintain genome integrity. Depending on the organism receiving the break, the genomic location of the DSB, and the cell-cycle phase in which it occurs, a DSB can be repaired by homologous recombination (HR), nonhomologous end-joining (NHEJ), or single-strand annealing (SSA). Two novel DSB repair assays were developed to determine the contributions of these repair pathways and to finely resolve repair event structures in Drosophila melanogaster. Rad51-dependent homologous recombination is the preferred DSB repair pathway in mitotically dividing cells, and the pathway choice between HR and SSA occurs after end resection and before Rad51-dependent strand invasion. HR events are associated with long gene conversion tracts and are both bidirectional and unidirectional, consistent with repair via the synthesis-dependent strand annealing pathway. Additionally, HR between diverged sequences is suppressed in Drosophila, similar to levels reported in human cells. Junction analyses of rare NHEJ events reveal that canonical NHEJ is utilized in this system.
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