Murayama Yasuto, Uhlmann Frank
Institute of Innovative Research, Tokyo Institute of Technology, M6-11, 2-12-1, Ookayama, Megro, Tokyo, 152-8550, Japan.
The Francis Crick Institute, Lincoln's Inn Fields Laboratory, London, UK.
Methods Mol Biol. 2017;1515:23-35. doi: 10.1007/978-1-4939-6545-8_2.
The cohesin complex is involved in a broad range of chromosomal biology, including DNA repair, gene transcription as well as sister chromatid cohesion. Cohesin is a large, ring-shaped protein complex and is thought to entrap DNA molecules inside of its ring. The unique DNA association is central to cohesin function and requires its ATPase and another heterodimer complex called the cohesin loader. Here we describe the biochemical reconstitution of topological cohesin loading onto DNA using the purified fission yeast cohesin proteins.
黏连蛋白复合体参与广泛的染色体生物学过程，包括DNA修复、基因转录以及姐妹染色单体黏连。黏连蛋白是一种大型环状蛋白质复合体，被认为能将DNA分子捕获在其环内。独特的DNA结合对于黏连蛋白的功能至关重要，并且需要其ATP酶和另一种称为黏连蛋白装载器的异二聚体复合体。在这里，我们描述了使用纯化的裂殖酵母黏连蛋白对拓扑黏连蛋白加载到DNA上进行生化重建。
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