Kelso Andrew A, Say Amanda F, Sharma Deepti, Ledford LeAnna L, Turchick Audrey, Saski Christopher A, King Ada V, Attaway Christopher C, Temesvari Lesly A, Sehorn Michael G
Department of Genetics and Biochemistry, Clemson University, Clemson, South Carolina, United States of America.
Clemson University Genomics and Computational Biology Laboratory, Institute for Translational Genomics, Clemson, South Carolina, United States of America.
PLoS One. 2015 Sep 30;10(9):e0139399. doi: 10.1371/journal.pone.0139399. eCollection 2015.
Meiosis depends on homologous recombination (HR) in most sexually reproducing organisms. Efficient meiotic HR requires the activity of the meiosis-specific recombinase, Dmc1. Previous work shows Dmc1 is expressed in Entamoeba histolytica, a eukaryotic parasite responsible for amoebiasis throughout the world, suggesting this organism undergoes meiosis. Here, we demonstrate Dmc1 protein is expressed in E. histolytica. We show that purified ehDmc1 forms presynaptic filaments and catalyzes ATP-dependent homologous DNA pairing and DNA strand exchange over at least several thousand base pairs. The DNA pairing and strand exchange activities are enhanced by the presence of calcium and the meiosis-specific recombination accessory factor, Hop2-Mnd1. In combination, calcium and Hop2-Mnd1 dramatically increase the rate of DNA strand exchange activity of ehDmc1. The biochemical system described herein provides a basis on which to better understand the role of ehDmc1 and other HR proteins in E. histolytica.
在大多数有性生殖生物中,减数分裂依赖于同源重组(HR)。高效的减数分裂HR需要减数分裂特异性重组酶Dmc1的活性。先前的研究表明,Dmc1在溶组织内阿米巴中表达,溶组织内阿米巴是一种在全球范围内引起阿米巴病的真核寄生虫,这表明该生物体经历减数分裂。在此,我们证明Dmc1蛋白在溶组织内阿米巴中表达。我们表明,纯化的ehDmc1形成突触前细丝,并催化至少数千个碱基对的ATP依赖性同源DNA配对和DNA链交换。钙和减数分裂特异性重组辅助因子Hop2-Mnd1的存在增强了DNA配对和链交换活性。钙和Hop2-Mnd1共同作用,显著提高了ehDmc1的DNA链交换活性速率。本文所述的生化系统为更好地理解ehDmc1和其他HR蛋白在溶组织内阿米巴中作用提供了基础。