Functional analyses of Pericentrin and Syne-2 interaction in ciliogenesis
Functional analyses of Pericentrin and Syne-2 interaction in ciliogenesis
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DOI：
10.1242/jcs.218487
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发表时间：
2018-08-01
影响因子：
4
通讯作者：
Giessl, Andreas
中科院分区：
文献类型：
--
作者：
Falk, Nathalie;Kessler, Kristin;Giessl, Andreas
Pericentrin（Pcnt）是一种多功能支架蛋白，人类PCNT基因突变与包括纤毛病在内的多种疾病相关。Pcnt在嗅觉感受器神经元纤毛发育中起着至关重要的作用，但其在连接光感受器纤毛中的功能尚不清楚。我们下调Pcnt在视网膜离体和在体内通过基于病毒的RNA干扰的方法来研究Pcnt在光感受器的功能。ShRNA介导的Pcnt敲低损害了连接纤毛和光感受器外段的发育，并导致核迁移缺陷。在蛋白质相互作用筛选中，我们发现外核膜蛋白Syne-2（也称为Nesprin-2）是光感受器中Pcnt的相互作用伴侣。Syne-2对小鼠感光细胞核的定位和纤毛发生早期中心体的迁移具有重要意义。CRISPR/Cas9介导的Syne-2敲除导致Pcnt过表达和错误定位以及纤毛发生缺陷。我们的研究结果表明，Pcnt-Syne-2复合物是重要的纤毛和视网膜发育过程中的外段形成，并在核迁移中发挥作用。
Pericentrin (Pcnt) is a multifunctional scaffold protein and mutations in the human PCNT gene are associated with several diseases, including ciliopathies. Pcnt plays a crucial role in ciliary development in olfactory receptor neurons, but its function in the photoreceptorconnecting cilium is unknown. We downregulated Pcnt in the retina ex vivo and in vivo via a virus-based RNA interference approach to study Pcnt function in photoreceptors. ShRNA-mediated knockdown of Pcnt impaired the development of the connecting cilium and the outer segment of photoreceptors, and caused a nuclear migration defect. In protein interaction screens, we found that the outer nuclear membrane protein Syne-2 (also known as Nesprin-2) is an interaction partner of Pcnt in photoreceptors. Syne-2 is important for positioning murine photoreceptor cell nuclei and for centrosomal migration during early ciliogenesis. CRISPR/Cas9mediated knockout of Syne-2 in cell culture led to an overexpression and mislocalization of Pcnt and to ciliogenesis defects. Our findings suggest that the Pcnt-Syne-2 complex is important for ciliogenesis and outer segment formation during retinal development and plays a role in nuclear migration.