Basello Davide A, Blažíková Michaela, Roithová Adriana, Hálová Martina, Radivojević Nenad, Neugebauer Karla M, Staněk David
Institute of Molecular Genetics, Czech Academy of Sciences , Prague, Czech Republic.
Faculty of Science, Charles University in Prague, Prague, Czech Republic.
J Cell Biol. 2025 Aug 4;224(8). doi: 10.1083/jcb.202309128. Epub 2025 Jun 25.
The Cajal body (CB) is a conserved non-membrane nuclear structure where several steps of small nuclear RNP particle (snRNP) biogenesis take place. It has been proposed that CB formation follows a liquid-liquid phase separation model, but this hypothesis has never been rigorously tested. Here, we applied live-cell imaging to show that the key CB assembly factor coilin is mobile within the CB, and we revealed a diffusion barrier that limits the coilin exchange between CBs and the nucleoplasm. We generated single aa mutations and demonstrated that RNA-dependent coilin oligomerization and coilin interaction with snRNP are essential for CB formation and maintenance. We applied these data to formulate a mathematical model that links the movement of coilin within the nucleoplasm, CB, and across the boundary with its oligomerization and snRNP binding. Our results illustrate CB as a structure dynamically responding to snRNP assembly and recycling.
卡哈尔体(CB)是一种保守的无膜核结构,小核核糖核蛋白颗粒(snRNP)生物发生的几个步骤在此进行。有人提出CB的形成遵循液-液相分离模型,但这一假设从未得到严格验证。在这里,我们应用活细胞成像表明,关键的CB组装因子卷曲螺旋蛋白在线粒体内是可移动的,并且我们揭示了一个扩散屏障,该屏障限制了卷曲螺旋蛋白在线粒体和核质之间的交换。我们产生了单个氨基酸突变,并证明了依赖RNA的卷曲螺旋蛋白寡聚化以及卷曲螺旋蛋白与snRNP的相互作用对于CB的形成和维持至关重要。我们应用这些数据构建了一个数学模型,该模型将卷曲螺旋蛋白在核质、线粒体内的运动及其跨越边界的运动与其寡聚化和snRNP结合联系起来。我们的结果表明,CB是一种对snRNP组装和循环动态响应的结构。