Department of Chemistry, National Taiwan University, Taiwan.
Department of Chemistry, National Taiwan Normal University, Taiwan.
Phys Chem Chem Phys. 2020 Jun 4;22(21):11928-11935. doi: 10.1039/d0cp01061h.
Polyamines are naturally occurring cationic molecules in cells. In addition to their roles in modulating gene expression and cell proliferation, they have been shown to stimulate DNA recombination. The molecular mechanism for stimulation is not clear. We utilized single-molecule tethered particle motion (TPM) experiments to investigate how polyamines stimulate RecA-mediated recombination. We showed that natural polyamines, spermine and spermidine, condense duplex DNA, but with different efficiencies. While ∼300 μM of spermine condenses 50% of duplex DNA, 2.0 mM of spermidine is required to achieve the same level of condensation. The condensation takes place in a stepwise manner, and is reversible upon removal of polyamines. We also showed that addition of polyamines stimulates the duplex capture activity of RecA filament and stabilizes the intermediates with longer dwell time. Through condensing duplex DNA and stabilizing the complex of RecA filaments and duplex DNA, polyamines stimulate the formation of functional intermediates by ∼20-fold, and promote recombination progression.
多胺是细胞中天然存在的阳离子分子。除了在调节基因表达和细胞增殖方面的作用外,它们还被证明能刺激 DNA 重组。刺激的分子机制尚不清楚。我们利用单分子系绳粒子运动(TPM)实验来研究多胺如何刺激 RecA 介导的重组。我们表明,天然多胺精胺和亚精胺使双链 DNA 凝聚,但效率不同。虽然 300μM 的精胺能使 50%的双链 DNA 凝聚,但需要 2.0mM 的亚精胺才能达到相同的凝聚水平。凝聚是逐步进行的,多胺去除后可逆转。我们还表明,多胺的添加刺激 RecA 丝的双链捕获活性,并稳定具有更长停留时间的中间体。通过使双链 DNA 凝聚并稳定 RecA 丝和双链 DNA 的复合物,多胺将功能性中间体的形成速度提高了约 20 倍,从而促进了重组的进行。