Topology-dependent anomalous dynamics of ring and linear DNA are sensitive to cytoskeleton crosslinking
Topology-dependent anomalous dynamics of ring and linear DNA are sensitive to cytoskeleton crosslinking
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DOI：
10.1126/sciadv.aay5912
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发表时间：
2019-10
期刊：
影响因子：
13.6
通讯作者：
Devynn M. Wulstein;Kathryn E. Regan;Jonathan Garamella;R. McGorty;R. Robertson-Anderson
中科院分区：
文献类型：
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作者：
Devynn M. Wulstein;Kathryn E. Regan;Jonathan Garamella;R. McGorty;R. Robertson-Anderson
拥挤的环状和线性DNA表现出不同的运输特性，受到细胞骨架交联的不同影响。细胞骨架拥挤在DNA分子通过细胞的扩散中起着关键作用，作为细胞内有效运输和构象稳定性的屏障，这些过程需要转染、病毒感染和基因治疗。在这里，我们阐明了线性和环状DNA分子在肌动蛋白和微管的纠缠和交联复合网络中扩散的传输特性和构象动力学。我们将单分子构象跟踪与差分动态显微镜相结合，揭示环状和线性DNA表现出出乎意料的不同运输特性，这些特性受到细胞骨架交联的不同影响。环状DNA线圈肿胀，并经历异质性和双相亚扩散，交联阻碍。相反，交联实际上促进了紧致线性链所表现出的单模亚扩散。我们的集体结果表明，细胞骨架细丝的瞬时穿线在环状DNA的动力学中起着关键作用，而细胞骨架的移动性决定了线性DNA的运输。
Crowded ring and linear DNA exhibit distinct transport properties that are influenced differently by cytoskeleton crosslinking. Cytoskeletal crowding plays a key role in the diffusion of DNA molecules through the cell, acting as a barrier to effective intracellular transport and conformational stability required for processes such as transfection, viral infection, and gene therapy. Here, we elucidate the transport properties and conformational dynamics of linear and ring DNA molecules diffusing through entangled and crosslinked composite networks of actin and microtubules. We couple single-molecule conformational tracking with differential dynamic microscopy to reveal that ring and linear DNA exhibit unexpectedly distinct transport properties that are influenced differently by cytoskeleton crosslinking. Ring DNA coils are swollen and undergo heterogeneous and biphasic subdiffusion that is hindered by crosslinking. Conversely, crosslinking actually facilitates the single-mode subdiffusion that compacted linear chains exhibit. Our collective results demonstrate that transient threading by cytoskeleton filaments plays a key role in the dynamics of ring DNA, whereas the mobility of the cytoskeleton dictates transport of linear DNA.