Preston Christine R, Engels William, Flores Carlos
Laboratory of Genetics, University of Wisconsin, Madison, Wisconsin 53706, USA.
Genetics. 2002 Jun;161(2):711-20. doi: 10.1093/genetics/161.2.711.
We show evidence that DNA double-strand breaks induced in the Drosophila germ line can be repaired very efficiently by the single-strand annealing (SSA) mechanism. A double-strand break was made between two copies of a 1290-bp direct repeat by mobilizing a P transposon. In >80% of the progeny that acquired this chromosome, repair resulted in loss of the P element and loss of one copy of the repeat, as observed in SSA. The frequency of this repair was much greater than seen for gene conversion using an allelic template, which is only approximately 7%. A similar structure, but with a smaller duplication of only 158 bp, also yielded SSA-like repair events, but at a reduced frequency, and gave rise to some products by repair pathways other than SSA. The 1290-bp repeats carried two sequence polymorphisms that were examined in the products. The allele nearest to a nick in the putative heteroduplex intermediate was lost most often. This bias is predicted by the SSA model, although other models could account for it. We conclude that SSA is the preferred repair pathway in Drosophila for DNA breaks between sequence repeats, and it competes with gene conversion by the synthesis-dependent strand annealing (SDSA) pathway.
我们提供的证据表明,果蝇生殖系中诱导产生的DNA双链断裂可通过单链退火(SSA)机制得到非常有效的修复。通过激活P转座子,在一个1290 bp直接重复序列的两个拷贝之间产生了一个双链断裂。在获得这条染色体的后代中,超过80%的修复导致P元件丢失和一个重复拷贝的丢失,这与SSA中观察到的情况一致。这种修复的频率远高于使用等位基因模板的基因转换,后者仅约为7%。一个类似的结构,但只有158 bp的较小重复,也产生了类似SSA的修复事件,但频率降低,并通过SSA以外的修复途径产生了一些产物。1290 bp的重复序列携带两个序列多态性,对产物进行了检测。最靠近假定异源双链中间体切口的等位基因丢失最为频繁。尽管其他模型也可以解释这种偏向性,但SSA模型对此进行了预测。我们得出结论,SSA是果蝇中修复序列重复之间DNA断裂的首选修复途径,并且它与合成依赖链退火(SDSA)途径介导的基因转换相互竞争。