Yoo Hyunjeong, Kim Taehyun, Ryu Sungjin, Ko Donghee, Kim Jeesoo, Choi Hee-Jung, Shin Yongdae, Rhee Kunsoo
Department of Biological Sciences, Seoul National University, Seoul 08826, Korea.
Department of Mechanical Engineering, Seoul National University, Seoul 08826, Korea.
J Cell Sci. 2025 May 15;138(10). doi: 10.1242/jcs.263542. Epub 2025 May 21.
The microtubule-organizing activity of centrosomes fluctuates during the cell cycle, reaching the highest levels at M phase. CEP215 (also known as CDK5RAP2) is a key pericentriolar material (PCM) protein for microtubule organization of the human centrosome. Here, we provide evidence that CEP215 exhibits a dynamically suppressed, solid-like state in interphase centrosomes, and becomes a more dynamic state in mitotic centrosomes. Specific interaction with PCNT, another centrosome protein, is crucial for diffusible molecular dynamicity of the CEP215 protein. We also found that the cluster formation activity of CEP215 is impaired in a light-inducible system when its coiled-coil domains (CCDs) are truncated. Defects in spindle pole assembly and spindle formation were accompanied in the cells whose CEP215 is replaced with the CCD-truncated mutants. Our results support the notion that the diffusible mobility of CEP215 is enhanced by both homotypic and heterotypic interactions among CCDs, especially at mitotic spindle poles. This work highlights that biophysical properties of the PCM proteins at the centrosomes fluctuate during the cell cycle.
中心体的微管组织活性在细胞周期中波动,在M期达到最高水平。CEP215(也称为CDK5RAP2)是人类中心体微管组织的关键中心粒外周物质(PCM)蛋白。在这里,我们提供证据表明,CEP215在间期中心体中表现出动态抑制的、类似固体的状态,而在有丝分裂中心体中变为更动态的状态。与另一种中心体蛋白PCNT的特异性相互作用对于CEP215蛋白的可扩散分子动态性至关重要。我们还发现,当CEP215的卷曲螺旋结构域(CCDs)被截断时,其在光诱导系统中的聚集形成活性受损。用CCDs截断突变体替代CEP215的细胞中伴随着纺锤极组装和纺锤体形成的缺陷。我们的结果支持这样一种观点,即CEP215的可扩散流动性通过CCDs之间的同型和异型相互作用而增强,特别是在有丝分裂纺锤极处。这项工作强调了中心体处PCM蛋白的生物物理性质在细胞周期中会发生波动。