Sen S, Karthikeyan G, Rao B J
Department of Biological Sciences, Tata Institute of Fundamental Research, Colaba, Mumbai, India.
Biochemistry. 2000 Aug 22;39(33):10196-206. doi: 10.1021/bi000753y.
Microsatellite repeats such as mono-, di-, and trinucleotides are highly abundant and viable targets for homologous recombination in the genome. However, if recombination ensues in such repetitive regions, they are intrinsically prone to frame misalignments during pairing and might eventually give rise to genetic instabilities. Suboptimally paired frames lead to an abrogation of branch migration at the junctions of mixed sequences and repeats, due to a heterologous register. If so, can recombination machinery rectify such misalignments in order to avoid subsequent arrest in branch migration? We analyzed Escherichia coli RecA, the universal prototype of a recombinase, for its pairing abilities across repeats. We used a complementary pairing assay to test whether RecA can mediate realignments of stochastically paired suboptimal frames to a maximally aligned register. Here, we demonstrate that RecA-single stranded DNA filament indeed facilitates such a realignment, probably by sliding the paired strands across mono- and di- as well as trinucleotide repeats. These realignments apparently have no net directional bias. Such a putative "motor" function of RecA seems to be ATP hydrolysis-dependent.
单核苷酸、二核苷酸和三核苷酸等微卫星重复序列在基因组中高度丰富,是同源重组的可行靶点。然而,如果在这些重复区域发生重组,它们在配对过程中本质上容易出现框架错位,最终可能导致基因不稳定。由于异源序列比对,次优配对框架会导致混合序列和重复序列交界处的分支迁移终止。如果是这样,重组机制能否纠正这种错位,以避免随后的分支迁移停滞?我们分析了通用重组酶原型——大肠杆菌RecA在重复序列上的配对能力。我们使用互补配对试验来测试RecA是否能将随机配对的次优框架重新排列到最大比对的序列比对中。在这里,我们证明RecA单链DNA细丝确实促进了这种重新排列,可能是通过使配对链在单核苷酸、二核苷酸和三核苷酸重复序列上滑动实现的。这些重新排列显然没有净方向偏差。RecA的这种假定“马达”功能似乎依赖于ATP水解。