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Elevated levels of intrachromosomal homologous recombination in Arabidopsis overexpressing the MIM gene

机译:过量表达MIM基因的拟南芥中染色体内同源重组水平升高

摘要

The Arabidopsis MIM gene encodes a protein belonging to the SMC family (structure maintenance of chromosomes) which is required for intrachromosomal homologous recombination (ICR). Both ICR and MIM gene expression are enhanced by DNA-damaging treatments, suggesting that MIM is a factor limiting DNA repair by homologous recombination (HR) under genotoxic stress. We tested this hypothesis by measuring the levels of recombination in the mim mutant under genotoxic stress, using methyl methanesulfonate. Although the mutant clearly showed diminished basal and induced levels of ICR, enhancement of ICR by DNA-damaging treatments was similar to that observed in the wild type. This suggests that the MIM gene product is required for DNA repair by HR, but is not critical for HR induction. To determine whether enhanced availability of MIM would increase basal HR levels in Arabidopsis, we examined ICR frequencies in transgenic Arabidopsis strains overexpressing the MIM gene after ectopic insertion of additional MIM copies. Two independent lines showed a twofold increase in ICR frequency relative to the wild type. Thus MIM is required for efficient ICR in plants, and its manipulation can be used to change homologous recombination frequencies. Since MIM is one of the components responsible for chromatin dynamics, our results suggest that the chromatin environment determines the frequency of homologous recombination.
机译:拟南芥MIM基因编码属于SMC家族(染色体的结构保持)的蛋白质,该蛋白质是染色体内同源重组(ICR)所必需的。 DNA损伤处理可增强ICR和MIM基因的表达,这表明MIM是在遗传毒性胁迫下通过同源重组(HR)限制DNA修复的因子。我们通过使用甲磺酸甲酯测量在遗传毒性胁迫下mim突变体中的重组水平来检验该假设。尽管该突变体明显显示出ICR的基础水平和诱导水平降低,但通过DNA损伤处理增强ICR与在野生型中观察到的相似。这表明MIM基因产物是HR修复DNA所必需的,但对于HR诱导并不是至关重要的。为了确定MIM可用性的提高是否会增加拟南芥的基础HR水平,我们在异位插入其他MIM拷贝后检查了过表达MIM基因的转基因拟南芥菌株中的ICR频率。两条独立的品系显示ICR频率相对于野生型增加了两倍。因此,MIM是植物中有效ICR所必需的,其操纵可用于改变同源重组频率。由于MIM是负责染色质动力学的成分之一,因此我们的结果表明,染色质环境决定了同源重组的频率。

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