State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, No. 19A Yuquan Road, Beijing 100049, China.
J Genet Genomics. 2017 Jan 20;44(1):39-49. doi: 10.1016/j.jgg.2016.11.005. Epub 2016 Dec 26.
The Golgi assembly pattern varies among cell types. In fibroblast cells, the Golgi apparatus concentrates around the centrosome that radiates microtubules; whereas in epithelial cells, whose microtubules are mainly noncentrosomal, the Golgi apparatus accumulates around the nucleus independently of centrosome. Little is known about the mechanisms behind such cell type-specific Golgi and microtubule organization. Here, we show that the microtubule minus-end binding protein Nezha/CAMSAP3 (calmodulin-regulated spectrin-associated protein 3) plays a role in translocation of Golgi vesicles in epithelial cells. This function of CAMSAP3 is supported by CG-NAP (centrosome and Golgi localized PKN-associated protein) through their binding. Depletion of either one of these proteins similarly induces fragmentation of Golgi membranes. Furthermore, we find that stathmin-dependent microtubule dynamics is graded along the radial axis of cells with highest activity at the perinuclear region, and inhibition of this gradient disrupts perinuclear distribution of the Golgi apparatus. We propose that the assembly of the Golgi apparatus in epithelial cells is induced by a multi-step process, which includes CAMSAP3-dependent Golgi vesicle clustering and graded microtubule dynamics.
高尔基体组装模式在细胞类型之间存在差异。在成纤维细胞中,高尔基体集中在放射微管的中心体周围;而在微管主要是非中心体的上皮细胞中,高尔基体独立于中心体聚集在核周围。对于这种细胞类型特异性的高尔基体和微管组织背后的机制知之甚少。在这里,我们表明微管负端结合蛋白 Nezha/CAMSAP3(钙调蛋白调节的 spectrin 相关蛋白 3)在上皮细胞中高尔基体小泡的易位中发挥作用。CG-NAP(中心体和高尔基体定位的 PKN 相关蛋白)通过与其结合支持 CAMSAP3 的此功能。这两种蛋白质之一的耗尽同样会诱导高尔基体膜的碎片化。此外,我们发现,stathmin 依赖性微管动力学沿着细胞的径向轴呈梯度分布,在核周区域活性最高,抑制该梯度会破坏高尔基体在核周的分布。我们提出,上皮细胞中高尔基体的组装是由一个多步骤过程诱导的,该过程包括 CAMSAP3 依赖性高尔基体小泡聚集和分级微管动力学。