Abstract:
Centriole biogenesis, as in most organelle assemblies, involves the sequential recruitment of sub-structural elements that will support its function. To uncover this process, we correlated the spatial location of 24 centriolar proteins with structural features using expansion microscopy. A time-series reconstruction of protein distributions throughout human procentriole assembly unveiled the molecular architecture of the centriole biogenesis steps. We found that the process initiates with the formation of a naked cartwheel devoid of microtubules. Next, the bloom phase progresses with microtubule blade assembly, concomitantly with radial separation and rapid cartwheel growth. In the subsequent elongation phase, the tubulin backbone grows linearly with the recruitment of the A-C linker, followed by proteins of the inner scaffold (IS). By following six structural modules, we modeled 4D assembly of the human centriole. Collectively, this work provides a framework to investigate the spatial and temporal assembly of large macromolecules.
摘要:
Centriole生物发生,和大多数细胞器装配一样,涉及顺序招募将支持其功能的子结构要素。为了揭开这个过程,我们使用扩展显微镜将24个中心摩尔蛋白的空间位置与结构特征相关联。整个人类原粒组装过程中蛋白质分布的时间序列重建揭示了中心粒生物生成步骤的分子结构。我们发现该过程始于没有微管的裸露车轮的形成。接下来,开花阶段随着微管叶片组装而进展,伴随着径向分离和快速的车轮生长。在随后的延伸阶段,微管蛋白骨架随着A-C接头的募集线性增长,其次是内部支架(IS)的蛋白质。通过遵循六个结构模块,我们模拟了人体中心的4D组装。总的来说,这项工作为研究大型大分子的时空组装提供了一个框架。