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  • Functional analyses of Pericentrin and Syne-2 interaction in ciliogenesis.

Functional analyses of Pericentrin and Syne-2 interaction in ciliogenesis.

Journal of cell science (2018-07-29)
Nathalie Falk, Kristin Kessler, Sinja-Fee Schramm, Karsten Boldt, Elvir Becirovic, Stylianos Michalakis, Hanna Regus-Leidig, Angelika A Noegel, Marius Ueffing, Christian T Thiel, Ronald Roepman, Johann Helmut Brandstätter, Andreas Gießl
ABSTRACT

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.

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