Functional analyses of Pericentrin and Syne-2/Nesprin-2 interaction in ciliogenesis
| Autor: | Nathalie Falk, Elvir Becirovic, Johann Helmut Brandstätter, Marius Ueffing, Kristin Kessler, Stylianos Michalakis, Christian Thiel, Hanna Regus-Leidig, Sinja-Fee Schramm, Karsten Boldt, Ronald Roepman, Andreas Gießl, Angelika A. Noegel |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Scaffold protein Gene knockdown Cilium Cell Biology Biology Ciliopathies Photoreceptor cell Cell biology 03 medical and health sciences 030104 developmental biology 0302 clinical medicine medicine.anatomical_structure RNA interference PCNT Ciliogenesis medicine sense organs 030217 neurology & neurosurgery |
| Zdroj: | Journal of Cell Science. |
| ISSN: | 1477-9137 0021-9533 |
| DOI: | 10.1242/jcs.218487 |
| Popis: | 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 photoreceptor-connecting 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/Cas9-mediated 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|