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Role of RAD51 in the Repair of MuDR-Induced Double-Strand Breaks in Maize (Zea mays L.)

机译:RAD51在修复MuDR诱导的玉米双链断裂中的作用(Zea mays L.)

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摘要

Rates of Mu transposon insertions and excisions are both high in late somatic cells of maize. In contrast, although high rates of insertions are observed in germinal cells, germinal excisions are recovered only rarely. Plants doubly homozygous for deletion alleles of rad51A1 and rad51A2 do not encode functional RAD51 protein (RAD51−). Approximately 1% of the gametes from RAD51+ plants that carry the MuDR-insertion allele a1-m5216 include at least partial deletions of MuDR and the a1 gene. The structures of these deletions suggest they arise via the repair of MuDR-induced double-strand breaks via nonhomologous end joining. In RAD51− plants these germinal deletions are recovered at rates that are at least 40-fold higher. These rates are not substantially affected by the presence or absence of an a1-containing homolog. Together, these findings indicate that in RAD51+ germinal cells MuDR-induced double-strand breaks (DSBs) are efficiently repaired via RAD51-directed homologous recombination with the sister chromatid. This suggests that RAD51− plants may offer an efficient means to generate deletion alleles for functional genomic studies. Additionally, the high proportion of Mu-active, RAD51− plants that exhibit severe developmental defects suggest that RAD51 plays a critical role in the repair of MuDR-induced DSBs early in vegetative development.
机译:在玉米的晚期体细胞中,Mu转座子的插入和切除率都很高。相比之下,尽管在生发细胞中观察到了很高的插入率,但生发切除很少被恢复。 rad51A1和rad51A2缺失等位基因的双纯合植物不编码功能性RAD51蛋白(RAD51-)。携带MuDR插入等位基因a1-m5216的RAD51 +植物的配子中,大约有1%包含MuDR和a1基因的至少部分缺失。这些缺失的结构表明,它们是通过非同源末端连接修复MuDR诱导的双链断裂而产生的。在RAD51-植物中,这些生发缺失的回收率至少高40倍。这些速率基本上不受存在或不存在含α1同系物的影响。总之,这些发现表明,在RAD51 +生发细胞中,MudR诱导的双链断裂(DSB)通过RAD51定向的与姐妹染色单体的同源重组而得到有效修复。这表明RAD51-植物可能为功能基因组研究提供了产生缺失等位基因的有效手段。此外,高比例的Mu-active RAD51-植物表现出严重的发育缺陷,表明RAD51在营养生长早期修复MuDR诱导的DSB中起着关键作用。

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