Waldvogel Sarah M, Goodson Steven D, Sehorn Michael G
Department of Genetics and Biochemistry, Clemson University, Clemson, SC, USA.
Eukaryotic Pathogens Innovation Center, Clemson University, Clemson, SC, USA.
Methods Mol Biol. 2019;1999:285-291. doi: 10.1007/978-1-4939-9500-4_19.
The meiosis-specific recombinase, DMC1, is important for the generation of haploids during meiosis. DMC1 forms a helical nucleoprotein filament on ssDNA overhangs located at the processed double-stranded DNA break. The DMC1 filament performs a search for homology in homologous chromosome. Once homology is located, the DMC1 filament strand invades the homologous chromosome forming a displacement loop (D-loop). These connections are needed for accurate segregation to occur later in meiosis. Because DMC1 requires numerous accessory factors and specific ionic conditions to participate in this DNA repair process, in vitro assays were developed to understand how these accessory factors influence the biochemical properties of hDMC1. This chapter describes a method that can be used to investigate the stability of the human DMC1 nucleoprotein filament under various conditions and provides insight into an important early stage in DNA double-strand break repair by homologous recombination during meiosis.
减数分裂特异性重组酶DMC1对于减数分裂过程中单倍体的产生至关重要。DMC1在位于已加工双链DNA断裂处的单链DNA突出端上形成螺旋核蛋白细丝。DMC1细丝在同源染色体中寻找同源性。一旦找到同源性,DMC1细丝链就会侵入同源染色体,形成置换环(D环)。这些连接是减数分裂后期准确分离所必需的。由于DMC1需要众多辅助因子和特定离子条件才能参与此DNA修复过程,因此开发了体外试验来了解这些辅助因子如何影响hDMC1的生化特性。本章描述了一种可用于研究人类DMC1核蛋白细丝在各种条件下稳定性的方法,并深入了解减数分裂期间通过同源重组进行DNA双链断裂修复的重要早期阶段。