Chenyang Gu, Shoji Takada, Giovani B. Brandani, Tsuyoshi Terakawa
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引用次数: 0
摘要
在真核细胞中,SMC 复合物凝聚素通过 DNA 环挤压介导染色质间期结构的形成。在此,我们试图利用分子动力学模拟研究其机制。为此,我们首先构建了SMC1、SMC3、STAG1和NIPBL等凝聚素亚基的氨基酸分辨率结构模型。通过用双链DNA分子模拟这些亚基,我们预测了每个亚基上的DNA结合斑块,并以这些斑块的解离率常数为代表,量化了这些斑块与DNA的亲和力。然后,我们构建了整个凝聚蛋白复合物的结构模型,并在该结构上绘制了预测的高亲和性 DNA 结合斑块。从预测的补丁的空间关系中,我们发现SMC1、SMC3、STAG1和NIPBL亚基上的多个补丁组成了一个DNA夹持补丁组。用双链 DNA 分子模拟整个复合体的结果表明,这个贴片组有利于 DNA 弯曲,并有助于在 SMC1 和 SMC3 亚基形成的凝聚素环中捕获 DNA 片段。在以前的研究中,这些已被确定为 DNA 环挤出的关键步骤。因此,本研究对之前提出的 DNA 环挤出机制中涉及的 DNA 结合位点进行了可实验检验的预测,并强调了附属亚基 STAG1 和 NIPBL 在该机制中的重要作用。
Molecular dynamics simulations of human cohesin subunits identify DNA binding sites and their potential roles in DNA loop extrusion
The SMC complex cohesin mediates interphase chromatin structural formation in eukaryotic cells through DNA loop extrusion. Here, we sought to investigate its mechanism using molecular dynamics simulations. To achieve this, we first constructed the amino-acid-residue-resolution structural models of the cohesin subunits, SMC1, SMC3, STAG1, and NIPBL. By simulating these subunits with double-stranded DNA molecules, we predicted DNA binding patches on each subunit and quantified the affinities of these patches to DNA using their dissociation rate constants as a proxy. Then, we constructed the structural model of the whole cohesin complex and mapped the predicted high-affinity DNA binding patches on the structure. From the spatial relations of the predicted patches, we identified that multiple patches on the SMC1, SMC3, STAG1, and NIPBL subunits form a DNA clamping patch group. The simulations of the whole complex with double-stranded DNA molecules suggest that this patch group facilitates DNA bending and helps capture a DNA segment in the cohesin ring formed by the SMC1 and SMC3 subunits. In previous studies, these have been identified as critical steps in DNA loop extrusion. Therefore, this study provides experimentally testable predictions of DNA binding sites implicated in previously proposed DNA loop extrusion mechanisms and highlights the essential roles of the accessory subunits STAG1 and NIPBL in the mechanism.