Department of Physics, University of Helsinki, Gustaf Hällströmin katu 2, P.O. Box 64, 00014, Helsinki, Finland.
Eur Biophys J. 2014 Mar;43(2-3):71-9. doi: 10.1007/s00249-013-0941-x. Epub 2014 Jan 30.
The simplified artificial environments in which highly complex biological systems are studied do not represent the crowded, dense, salty, and dynamic environment inside the living cell. Consequently, it is important to investigate the effect of crowding agents on DNA. We used a dual-trap optical tweezers instrument to perform force spectroscopy experiments at pull speeds ranging from 0.3 to 270 μm/s on single dsDNA molecules in the presence of poly(ethylene glycol) (PEG) and monovalent salt. PEG of sizes 1,500 and 4,000 Da condensed DNA, and force-extension data contained a force plateau at approximately 1 pN. The level of the force plateau increased with increasing pull speed. During slow pulling the dissipated work increased linearly with pull speed. The calculated friction coefficient did not depend on amount of DNA incorporated in the condensate, indicating internal friction is independent of the condensate size. PEG300 had no effect on the dsDNA force-extension curve. The force plateau implies that condensation induced by crowding agents resembles condensation induced by multivalent cations.
在高度复杂的生物系统中研究的简化人工环境并不能代表活细胞内拥挤、密集、咸和动态的环境。因此,研究拥挤剂对 DNA 的影响很重要。我们使用双阱光镊仪器在聚乙二醇(PEG)和单价盐存在的情况下,以 0.3 到 270 μm/s 的拉伸速度对单个双链 DNA 分子进行力谱实验。大小为 1500 和 4000 Da 的 PEG 使 DNA 凝聚,力-延伸数据在大约 1 pN 处包含一个力平台。力平台的水平随拉伸速度的增加而增加。在缓慢拉伸过程中,耗散功随拉伸速度线性增加。计算出的摩擦系数不依赖于凝聚物中包含的 DNA 量,表明内摩擦与凝聚物的大小无关。PEG300 对 dsDNA 力-延伸曲线没有影响。力平台表明,拥挤剂诱导的凝聚类似于多价阳离子诱导的凝聚。