The Medical Research Council Cancer Unit, University of Cambridge, Hills Road, Cambridge, United Kingdom.
Photonics Group, Department of Physics, Imperial College London, London, United Kingdom.
PLoS Biol. 2022 Oct 25;20(10):e3001854. doi: 10.1371/journal.pbio.3001854. eCollection 2022 Oct.
Centrioles are non-membrane-bound organelles that participate in fundamental cellular processes through their ability to form physical contacts with other structures. During interphase, two mature centrioles can associate to form a single centrosome-a phenomenon known as centrosome cohesion. Centrosome cohesion is important for processes such as cell migration, and yet how it is maintained is unclear. Current models indicate that pericentriolar fibres termed rootlets, also known as the centrosome linker, entangle to maintain centriole proximity. Here, I uncover a centriole-centriole contact site and mechanism of centrosome cohesion based on coalescence of the proximal centriole component cNap1. Using live-cell imaging of endogenously tagged cNap1, I show that proximal centrioles form dynamic contacts in response to physical force from the cytoskeleton. Expansion microscopy reveals that cNap1 bridges between these contact sites, physically linking proximal centrioles on the nanoscale. Fluorescence correlation spectroscopy (FCS)-calibrated imaging shows that cNap1 accumulates at nearly micromolar concentrations on proximal centrioles, corresponding to a few hundred protein copy numbers. When ectopically tethered to organelles such as lysosomes, cNap1 forms viscous and cohesive assemblies that promote organelle spatial proximity. These results suggest a mechanism of centrosome cohesion by cNap1 at the proximal centriole and illustrate how a non-membrane-bound organelle forms organelle contact sites.
中心体是无膜结合的细胞器,通过与其他结构形成物理接触,参与基本的细胞过程。在细胞间期,两个成熟的中心体可以结合形成一个单一的中心体——这种现象称为中心体凝聚。中心体凝聚对于细胞迁移等过程很重要,但它是如何维持的还不清楚。目前的模型表明,称为根丝的中心体周围纤维,也称为中心体连接蛋白,缠绕在一起以保持中心体的接近。在这里,我基于近端中心体组件 cNap1 的凝聚,揭示了中心体-中心体接触位点和中心体凝聚的机制。使用内源性标记的 cNap1 的活细胞成像,我表明近端中心体在细胞骨架的物理力作用下形成动态接触。扩展显微镜显示,cNap1 在这些接触位点之间桥接,在纳米尺度上物理连接近端中心体。荧光相关光谱(FCS)校准成像表明,cNap1 在近端中心体上以近微摩尔浓度积累,对应于几百个蛋白质拷贝数。当异位固定在溶酶体等细胞器上时,cNap1 形成粘性和凝聚的组装体,促进细胞器的空间接近。这些结果表明 cNap1 在近端中心体处的中心体凝聚机制,并说明了无膜结合细胞器如何形成细胞器接触位点。