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首页> 外文期刊>Molecular and Cellular Biology >The Drosophila melanogaster DmRAD54 Gene Plays a Crucial Role in Double-Strand Break Repair after P-Element Excision and Acts Synergistically with Ku70 in the Repair of X-Ray Damage
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The Drosophila melanogaster DmRAD54 Gene Plays a Crucial Role in Double-Strand Break Repair after P-Element Excision and Acts Synergistically with Ku70 in the Repair of X-Ray Damage

机译：果蝇DmRAD54基因在P元素切除后在双链断裂修复中起关键作用，并与Ku70协同作用修复X射线损伤

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The RAD54 gene has an essential role in the repair of double-strand breaks (DSBs) via homologous recombination in yeast as well as in higher eukaryotes. A Drosophila melanogasterstrain deficient in the RAD54 homolog DmRAD54is characterized by increased X-ray and methyl methanesulfonate (MMS) sensitivity. In addition, DmRAD54 is involved in the repair of DNA interstrand cross-links, as is shown here. However, whereas X-ray-induced loss-of-heterozygosity (LOH) events were completely absent in DmRAD54 ?/? flies, treatment with cross-linking agents or MMS resulted in only a slight reduction in LOH events in comparison with those in wild-type flies. To investigate the relative contributions of recombinational repair and nonhomologous end joining in DSB repair, aDmRad54 ?/?/DmKu70 ?/?double mutant was generated. Compared with both single mutants, a strong synergistic increase in X-ray sensitivity was observed in the double mutant. No similar increase in sensitivity was seen after treatment with MMS. Apparently, the two DSB repair pathways overlap much less in the repair of MMS-induced lesions than in that of X-ray-induced lesions. Excision of P transposable elements inDrosophila involves the formation of site-specific DSBs. In the absence of the DmRAD54 gene product, no male flies could be recovered after the excision of a single P element and the survival of females was reduced to 10% compared to that of wild-type flies. P-element excision involves the formation of two DSBs which have identical 3′ overhangs of 17 nucleotides. The crucial role of homologous recombination in the repair of these DSBs may be related to the very specific nature of the breaks.
机译： RAD54 基因在酵母以及高级真核生物中通过同源重组在修复双链断裂（DSB）中起着至关重要的作用。缺乏 RAD54 同源物 DmRAD54 的果蝇（Drosophila melanogaster）菌株的特征在于X射线和甲烷磺酸甲酯（MMS）的敏感性增加。另外，如下所示， DmRAD54 参与DNA链间交联的修复。但是，虽然在 DmRAD54 ？/？果蝇中完全没有X射线引起的杂合性丧失（LOH）事件，但是却产生了使用交联剂或MMS的治疗与野生型果蝇相比，LOH事件仅略有减少。要研究重组修复和非同源末端连接在DSB修复中的相对贡献，请使用 DmRad54 ？/？ / DmKu70 ？/？ 双重突变体已产生。与两个单突变体相比，在双突变体中观察到X射线敏感性的强烈协同增加。用MMS治疗后，没有发现敏感性的类似增加。显然，在MMS诱发的病变的修复中，两条DSB修复途径的重叠要比X射线诱发的病变的重叠少得多。果蝇中P转座子的切除涉及位点特异性DSB的形成。在缺乏 DmRAD54 基因产物的情况下，单个P元素被切除后，雄性蝇无法恢复，雌性存活率比野生型蝇降低了10％。 P元素切除涉及形成两个具有17个核苷酸相同3'突出端的DSB。同源重组在修复这些DSB中的关键作用可能与断裂的非常特殊的性质有关。

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