Centriolar satellites are required for efficient ciliogenesis and ciliary content regulation
Centriolar satellites are required for efficient ciliogenesis and ciliary content regulation
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DOI：
10.15252/embr.201947723
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发表时间：
2019-06-01
期刊：
影响因子：
7.7
通讯作者：
Firat-Karalar, Elif N.
中科院分区：
文献类型：
--
作者：
Odabasi, Ezgi;Gul, Seref;Firat-Karalar, Elif N.
着丝粒卫星在脊椎动物细胞中普遍存在。它们最近成为中心体/纤毛生物发生的关键调节因子，它们的突变与纤毛病有关。然而，对其确切的功能和作用机制仍然知之甚少。在这里，我们通过CRISPR/Cas9介导的PCM 1缺失产生了缺乏卫星的肾上皮细胞系（IMCD 3），并研究了卫星丢失的细胞和分子后果。缺乏卫星的细胞仍然形成全长纤毛，但数量显着减少，在中心体和细胞水平的关键纤毛发生因子的变化。使用这些细胞，我们确定了新的纤毛功能的卫星，如调节纤毛内容，刺猬信号，和上皮细胞组织在三维培养。然而，卫星的其他功能，即增殖，细胞周期进程，和中心粒复制，在这些细胞中不受影响。定量转录组学和蛋白质组学分析表明，卫星的损失几乎不影响转录，但显着改变蛋白质组。重要的是，中心体蛋白质组在缺乏卫星的细胞中大多保持不变。总之，我们的研究结果确定中心粒卫星作为有效的纤毛组装和功能的监管机构，并提供深入了解纤毛病变的疾病机制。
Centriolar satellites are ubiquitous in vertebrate cells. They have recently emerged as key regulators of centrosome/cilium biogenesis, and their mutations are linked to ciliopathies. However, their precise functions and mechanisms of action remain poorly understood. Here, we generated a kidney epithelial cell line (IMCD3) lacking satellites by CRISPR/Cas9-mediated PCM1 deletion and investigated the cellular and molecular consequences of satellite loss. Cells lacking satellites still formed full-length cilia but at significantly lower numbers, with changes in the centrosomal and cellular levels of key ciliogenesis factors. Using these cells, we identified new ciliary functions of satellites such as regulation of ciliary content, Hedgehog signaling, and epithelial cell organization in three-dimensional cultures. However, other functions of satellites, namely proliferation, cell cycle progression, and centriole duplication, were unaffected in these cells. Quantitative transcriptomic and proteomic profiling revealed that loss of satellites affects transcription scarcely, but significantly alters the proteome. Importantly, the centrosome proteome mostly remains unaltered in the cells lacking satellites. Together, our findings identify centriolar satellites as regulators of efficient cilium assembly and function and provide insight into disease mechanisms of ciliopathies.