LaFave Matthew C, Andersen Sabrina L, Stoffregen Eric P, Holsclaw Julie K, Kohl Kathryn P, Overton Lewis J, Sekelsky Jeff
Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, North Carolina 27599.
Genetics. 2014 Jan;196(1):107-18. doi: 10.1534/genetics.113.158618. Epub 2013 Oct 30.
The Bloom syndrome helicase, BLM, has numerous functions that prevent mitotic crossovers. We used unique features of Drosophila melanogaster to investigate origins and properties of mitotic crossovers that occur when BLM is absent. Induction of lesions that block replication forks increased crossover frequencies, consistent with functions for BLM in responding to fork blockage. In contrast, treatment with hydroxyurea, which stalls forks, did not elevate crossovers, even though mutants lacking BLM are sensitive to killing by this agent. To learn about sources of spontaneous recombination, we mapped mitotic crossovers in mutants lacking BLM. In the male germline, irradiation-induced crossovers were distributed randomly across the euchromatin, but spontaneous crossovers were nonrandom. We suggest that regions of the genome with a high frequency of mitotic crossovers may be analogous to common fragile sites in the human genome. Interestingly, in the male germline there is a paucity of crossovers in the interval that spans the pericentric heterochromatin, but in the female germline this interval is more prone to crossing over. Finally, our system allowed us to recover pairs of reciprocal crossover chromosomes. Sequencing of these revealed the existence of gene conversion tracts and did not provide any evidence for mutations associated with crossovers. These findings provide important new insights into sources and structures of mitotic crossovers and functions of BLM helicase.
布卢姆综合征解旋酶BLM具有多种防止有丝分裂交换的功能。我们利用黑腹果蝇的独特特征来研究在缺乏BLM时发生的有丝分裂交换的起源和特性。诱导阻断复制叉的损伤会增加交换频率,这与BLM在应对叉阻断方面的功能一致。相比之下,用羟基脲处理使复制叉停滞,即使缺乏BLM的突变体对该试剂杀伤敏感,也不会提高交换频率。为了了解自发重组的来源,我们在缺乏BLM的突变体中绘制了有丝分裂交换图谱。在雄性生殖系中,辐射诱导的交换随机分布在常染色质上,但自发交换是非随机的。我们认为,有丝分裂交换频率高的基因组区域可能类似于人类基因组中的常见脆性位点。有趣的是,在雄性生殖系中,跨越着丝粒周围异染色质的区间交换很少,但在雌性生殖系中,这个区间更容易发生交换。最后,我们的系统使我们能够回收相互交换的染色体对。对这些染色体的测序揭示了基因转换序列的存在,并且没有提供与交换相关的突变的任何证据。这些发现为有丝分裂交换的来源和结构以及BLM解旋酶的功能提供了重要的新见解。