Bonato Andrea, Jang Jae-Won, Kim Do-Gyun, Moon Kyoung-Wook, Michieletto Davide, Ryu Je-Kyung
Department of Physics, University of Strathclyde, Glasgow G4 0NG, United Kingdom.
Interdisciplinary Program in Computational Science, Seoul National University, Seoul 08826, South Korea.
Nucleic Acids Res. 2025 Jul 19;53(14). doi: 10.1093/nar/gkaf725.
DNA loop formation by structural maintenance of chromosome (SMC) proteins, including cohesin, condensin, and the SMC5/6 complex, plays a pivotal role in genome organization. Despite its importance, the molecular mechanism underlying SMC-mediated loop formation, particularly how these complexes achieve persistent directionality (rectification) while minimizing backward steps during the formation of large loops, remains poorly understood. Here, we use atomic force microscopy (AFM) and computational simulation to uncover a key geometric feature of the yeast condensin SMC complex enabling rectified loop growth. Using AFM, we demonstrate that the hinge domain of yeast condensin exhibits a directional bias, extending orthogonally to the bound DNA and sampling an anisotropic region of space around the protein complex. By accounting for the geometric constraint on the hinge-mediated DNA-capture step, we computationally show that loop growth can spontaneously self-rectify. In contrast, an SMC model with broken detailed balance and isotropic search instead exhibited substantial loop shrinkage and random-walk-like behaviour. These findings reveal an overlooked, and potentially broadly conserved, anisotropic DNA capture mechanism through which SMC complexes form and stabilize DNA loops in vivo, in turn providing novel insights into the physical principles governing genome organization.
包括黏连蛋白、凝聚素和SMC5/6复合物在内的染色体结构维持(SMC)蛋白介导的DNA环形成在基因组组织中起着关键作用。尽管其很重要,但SMC介导的环形成的分子机制,特别是这些复合物在形成大环时如何在尽量减少向后步骤的同时实现持续的方向性(整流),仍然知之甚少。在这里,我们使用原子力显微镜(AFM)和计算模拟来揭示酵母凝聚素SMC复合物能够实现整流环生长的一个关键几何特征。使用AFM,我们证明酵母凝聚素的铰链结构域表现出方向偏向性,垂直于结合的DNA延伸并在蛋白质复合物周围的空间中采样一个各向异性区域。通过考虑铰链介导的DNA捕获步骤的几何约束,我们通过计算表明环生长可以自发地自我整流。相比之下,一个具有破坏的细致平衡和各向同性搜索的SMC模型则表现出大量的环收缩和类似随机游走的行为。这些发现揭示了一种被忽视的、可能广泛保守的各向异性DNA捕获机制,通过该机制,SMC复合物在体内形成并稳定DNA环,进而为支配基因组组织的物理原理提供了新的见解。