Centre for Chromosome Biology, School of Natural Sciences, National University of Ireland Galway, Galway, Ireland.
Institute of Biochemistry and Molecular Biology, Ulm University, Ulm, Germany.
J Cell Biol. 2018 Apr 2;217(4):1205-1215. doi: 10.1083/jcb.201706095. Epub 2018 Feb 13.
The BRCA2 interactor, centrobin, is a centrosomal protein that has been implicated in centriole duplication and microtubule stability. We used genome editing to ablate in hTERT-RPE1 cells and observed an increased frequency of monocentriolar and acentriolar cells. Using a novel monoclonal antibody, we found that centrobin primarily localizes to daughter centrioles but also associates with mother centrioles upon serum starvation. Strikingly, centrobin loss abrogated primary ciliation upon serum starvation. Ultrastructural analysis of centrobin nulls revealed defective axonemal extension after mother centriole docking. Ciliogenesis required a C-terminal portion of centrobin that interacts with CP110 and tubulin. We also depleted centrobin in zebrafish embryos to explore its roles in an entire organism. Centrobin-depleted embryos showed microcephaly, with curved and shorter bodies, along with marked defects in laterality control, morphological features that indicate ciliary dysfunction. Our data identify new roles for centrobin as a positive regulator of vertebrate ciliogenesis.
BRCA2 相互作用蛋白 centrobin 是一种中心体蛋白，它与中心体复制和微管稳定性有关。我们使用基因组编辑技术敲除了 hTERT-RPE1 细胞中的 centrobin，并观察到单中心体和无中心体细胞的频率增加。使用一种新型单克隆抗体，我们发现 centrobin 主要定位于子中心体，但在血清饥饿时也与母中心体结合。引人注目的是，centrobin 的缺失消除了血清饥饿时的初级纤毛发生。centrobin 缺失体的超微结构分析显示，在母中心体对接后轴丝延伸缺陷。纤毛发生需要与 CP110 和微管相互作用的 centrobin 的 C 末端部分。我们还在斑马鱼胚胎中耗尽 centrobin，以探索其在整个生物体中的作用。centrobin 耗尽的胚胎表现出小头畸形，身体弯曲且较短，同时存在明显的左右不对称控制缺陷，这些形态特征表明纤毛功能障碍。我们的数据确定了 centrobin 作为脊椎动物纤毛发生的正调节剂的新作用。
