Super-resolution microscopy reveals coupling between mammalian centriole subdistal appendages and distal appendages
Super-resolution microscopy reveals coupling between mammalian centriole subdistal appendages and distal appendages
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DOI：
10.7554/elife.53580
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发表时间：
2020-04-03
期刊：
影响因子：
7.7
通讯作者：
Liao, Jung-Chi
中科院分区：
文献类型：
--
作者：
Chong, Weng Man;Wang, Won-Jing;Liao, Jung-Chi
近远侧附属物（Subdistal appendages，sDAP）是脊椎动物中在远侧附属物（distal appendages，DAP）近侧观察到的中心粒元件。尽管sDAP明显存在，但对它们的结构和功能的理解仍然难以捉摸。在这里，通过结合超分辨定位分析和CRISPR-Cas9遗传扰动，我们发现尽管DAP和sDAP分别主要负责纤毛发生和微管锚定中的不同功能，但一个元件的存在实际上会影响另一个元件的定位。具体而言，我们发现ODF 2和CEP 89的双层，其中它们的定位受到DAP和sDAP完整性的差异调节。DAP耗尽松弛sDAP蛋白ninein的纵向占据以覆盖DAP区域，这意味着DAP在sDAP定位中的作用。去除sDAP改变了中心体γ-微管蛋白的远端边界，说明了sDAP的新作用。总之，我们的研究结果为sDAP提供了一个架构框架，揭示了功能理解，令人惊讶地揭示了DAP和sDAP之间的耦合。
Subdistal appendages (sDAPs) are centriolar elements that are observed proximal to the distal appendages (DAPs) in vertebrates. Despite the obvious presence of sDAPs, structural and functional understanding of them remains elusive. Here, by combining super-resolved localization analysis and CRISPR-Cas9 genetic perturbation, we find that although DAPs and sDAPs are primarily responsible for distinct functions in ciliogenesis and microtubule anchoring, respectively, the presence of one element actually affects the positioning of the other. Specifically, we find dual layers of both ODF2 and CEP89, where their localizations are differentially regulated by DAP and sDAP integrity. DAP depletion relaxes longitudinal occupancy of sDAP protein ninein to cover the DAP region, implying a role of DAPs in sDAP positioning. Removing sDAPs alter the distal border of centrosomal gamma-tubulins, illustrating a new role of sDAPs. Together, our results provide an architectural framework for sDAPs that sheds light on functional understanding, surprisingly revealing coupling between DAPs and sDAPs.