Functional characterization of the protein Pericentrin (Kendrin) and identification of new interaction partners in the mammal retina
Final Report Abstract
In our study, we set out to examine the cellular and molecular functions of the multifunctional scaffold protein Pcnt in photoreceptors, the ciliated sensory neurons of the retina, by functionally analyzing the interactions of Pcnt with ciliary as well as centrosomal Pcnt interaction partners. Since we were not able to create a photoreceptor-specific conditional knockout model of Pcnt from embryonic stem (ES) cells in time, we had to limit our analyses to cell culture and other KO animals (like the Syne-2 KO mouse). The key findings of our study were: Knockdown and knockout findings indicate that a Pcnt plays a big role in cell-cycle and in ciliogenesis in cell culture and in the retina. - In our big interaction screens, we identified a group of Pcnt interaction partners with known functions at/in the nucleus, among which was Syne-2, a large 800 kDa protein. - We were able to show in studies that the mouse model Nesprin-2∆ABD is not suitable for studying the interaction between Syne-2 and Pcnt in vivo. - CRISPR/Cas9-mediated knockout of Syne-2 in cell culture led to an overexpression and mislocalization of Pcnt and to ciliogenesis defects. - The interaction between Pcnt and Syne-2 is required for ciliogenesis and ciliary length control, and seems to play a role in centrosomal-mediated nuclear migration. - One of our best candidate in the interaction screens seemed to be the protein CFAP298 (cilia and flagella associated protein 298) or also known as C21orf59 or kurly. It is expressed in ciliated tissue and further analysis will bring more light in the function of CFAP298 and Pcnt.
Publications
- (2018) Functional analyses of Pericentrin and Syne-2 interaction in ciliogenesis. J Cell Sci;131(16)
Falk N, Kessler K, Schramm SF, Boldt K, Becirovic E, Michalakis S, Regus-Leidig H, Noegel AA, Ueffing M, Thiel CT, Roepman R, Brandstätter JH, Gießl A
(See online at https://doi.org/10.1242/jcs.218487) - (2019) Lack of a Retinal Phenotype in a Syne-2/Nesprin-2 Knockout Mouse Model. Cells; 8(10)
Falk N, Joachimsthaler A, Kessler K, Lux UT, Noegel AA, Kremers J, Brandstätter JH, Gießl A
(See online at https://doi.org/10.3390/cells8101238)
