Bilichak Andriy, Yao Youli, Titov Viktor, Golubov Andrey, Kovalchuk Igor
Department of Biological Sciences, University of Lethbridge, University Drive 4401, Lethbridge, AB, T1K 3M4, Canada,
Plant Cell Rep. 2014 Jun;33(6):979-91. doi: 10.1007/s00299-014-1580-0. Epub 2014 Feb 20.
Plant XPD homolog UVH6 is the protein involved in the repair of strand breaks, and the excision repair and uvh6 mutant is not impaired in transgenerational increase in HRF. While analyzing the transgenerational response to stress in plants, we found that the promoter and gene body of Arabidopsis thaliana (Arabidopsis) XPD homolog UVH6 underwent hypomethylation and showed an increase in the level of transcript. Here, we analyzed the mutant of this gene, uvh6-1, by crossing it to two different reporter lines: one which allows for analysis of homologous recombination frequency (HRF) and another which makes it possible to analyze the frequency of point mutations. We observed that uvh6-1 plants exhibited lower rate of spontaneous homologous recombination but higher frequencies of spontaneous point mutations. The analysis of strand breaks using ROPS and Comet assays showed that the mutant had a much higher level of strand breaks at non-induced conditions. Exposure to stresses such as UVC, heat, cold, flood and drought showed that the mutant was not impaired in an increase in somatic HRF. The analysis of spontaneous HRF in the progeny of control plants compared to that of the progeny of stressed plants demonstrated that uvh6-1 was mildly affected in response to temperature, UV and drought. Our data suggest that UVH6 may be involved in the repair of strand breaks and excision repair, but it is unlikely that UVH6 is required for transgenerational increase in HRF.
植物XPD同源物UVH6是参与链断裂修复和切除修复的蛋白质,uvh6突变体在HRF的跨代增加方面没有受损。在分析植物对胁迫的跨代反应时,我们发现拟南芥XPD同源物UVH6的启动子和基因体发生了低甲基化,并显示转录水平增加。在这里,我们通过将该基因的突变体uvh6-1与两个不同的报告系杂交来分析它:一个允许分析同源重组频率(HRF),另一个使得分析点突变频率成为可能。我们观察到uvh6-1植株表现出自发同源重组率较低,但自发点突变频率较高。使用ROPS和彗星试验对链断裂进行分析表明,该突变体在非诱导条件下具有更高水平的链断裂。暴露于UVC、热、冷、洪水和干旱等胁迫下表明,该突变体在体细胞HRF增加方面没有受损。与胁迫植株后代相比,对照植株后代自发HRF的分析表明,uvh6-1对温度、紫外线和干旱的反应受到轻微影响。我们的数据表明,UVH6可能参与链断裂修复和切除修复,但UVH6不太可能是HRF跨代增加所必需的。