Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230026, PR China.
J Struct Biol. 2011 Apr;174(1):213-22. doi: 10.1016/j.jsb.2010.11.016. Epub 2010 Nov 18.
Prp20p is the homolog of mammalian RCC1 (regulator of chromosome condensation 1) in Saccharomyces cerevisiae, which acts as the guanine nucleotide exchange factor (GEF) for Gsp1p (yeast Ran). Prp20p plays multiple roles in mRNA metabolism, nucleocytoplasmic transport and mitosis regulation. Prp20p also functions as a linker between chromatin and nuclear pore complex (NPC) which regulates the NPC-mediated boundary activity (BA). Prp20p contains an N-terminal nuclear localization signal (NLS) and a typical RCC1-like domain (RLD). Here we present the 1.9Å crystal structure of the RCC1-like domain of Prp20p, which exhibits a classical seven-bladed β-propeller. We also proved that the additional β-wedge in Prp20p is essential for the interaction between Prp20p and Gsp1p. Based on this structure, we built a complex model of Prp20p and Gsp1p which was optimized by molecular dynamics (MD) simulations. Our model reveals that Prp20p and RCC1 share similar Ran GTPase binding mode. In addition, we also studied the histone-binding property of Prp20p in vitro.
Prp20p 是酿酒酵母中哺乳动物 RCC1（染色体凝聚调节剂 1）的同源物，作为 Gsp1p（酵母 Ran）的鸟嘌呤核苷酸交换因子（GEF）发挥作用。Prp20p 在 mRNA 代谢、核质转运和有丝分裂调节中发挥多种作用。Prp20p 还作为染色质和核孔复合物（NPC）之间的连接物发挥作用，调节 NPC 介导的边界活性（BA）。Prp20p 含有一个 N 端核定位信号（NLS）和一个典型的 RCC1 样结构域（RLD）。我们在此展示了 Prp20p 的 RCC1 样结构域的 1.9Å 晶体结构，其呈现出典型的七叶β-螺旋桨结构。我们还证明了 Prp20p 中的附加β-楔形结构对于 Prp20p 和 Gsp1p 之间的相互作用至关重要。基于此结构，我们构建了 Prp20p 和 Gsp1p 的复合物模型，并通过分子动力学（MD）模拟对其进行了优化。我们的模型表明，Prp20p 和 RCC1 具有相似的 Ran GTPase 结合模式。此外，我们还研究了 Prp20p 在体外的组蛋白结合特性。
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