Shukla Arun Kumar, Roy Kunal B
Centre for Biotechnology, Jawaharlal Nehru University, New Mehrauli Road, New Delhi 110067, India.
Biol Chem. 2006 Mar;387(3):251-6. doi: 10.1515/BC.2006.033.
We have recently reported that a GC-rich palindromic repeat sequence presumably adopts a stable fold-back tetraplex DNA structure under supercoiling. To establish the biological significance of this structure, we inserted this sequence between two direct repeat sequences, separated by 200 bp, in a plasmid. We then investigated the effect of this sequence on homologous recombination events. Here we report that the putative fold-back DNA tetraplex structure induces homologous recombination between direct repeat sequences. Interestingly, this recombination event is independent of recA, a major driving force for homologous recombination. We think that the fold-back structure forces the repeat sequences to come into close proximity and therefore leads to strand exchange. Although triplex-induced recombination has been well documented, our results for the first time directly establish the potential of a tetraplex structure to induce recA-independent homologous recombination in vivo. This finding might have a significant implication for site-directed gene deletion in the context of the correction of genetic defects.
我们最近报道,富含鸟嘌呤的回文重复序列在超螺旋状态下可能会形成稳定的回折四链体DNA结构。为了确定这种结构的生物学意义,我们将该序列插入到质粒中两个间隔200 bp的同向重复序列之间。然后我们研究了该序列对同源重组事件的影响。在此我们报道,假定的回折DNA四链体结构可诱导同向重复序列之间的同源重组。有趣的是,这种重组事件不依赖于recA,而recA是同源重组的主要驱动力。我们认为,回折结构迫使重复序列紧密靠近,从而导致链交换。尽管三链体诱导的重组已有充分记载,但我们的结果首次直接证实了四链体结构在体内诱导不依赖recA的同源重组的潜力。这一发现可能对纠正遗传缺陷背景下的定点基因缺失具有重要意义。