TurboID proximity interactome mapping reveals NR2E3 association with AP-1 and retinal developmental complexes
De Bruycker, E.; Zwaenepoel, O.; Moschonas, G. D.; De Sutter, D.; D'haene, E.; Matton, C.; Vandevelde, L.; Van De Velde, J.; Mahieu, Q.; Dufour, S.; Van Haver, D.; Delhaye, L.; Devos, S.; Coppieters, F.; Eyckerman, S.; Gettemans, J.; De Baere, E.
Show abstract
The photoreceptor-specific nuclear receptor NR2E3 is a key transcription factor in the retinal transcriptional network, essential in photoreceptor cell fate and maintenance. Various biallelic loss-of-function variants lead to autosomal recessive inherited retinal disease (IRD), while a single missense variant G56R has been identified to cause autosomal dominant retinitis pigmentosa (adRP). However, a comprehensive understanding of the interactome of NR2E3 remains elusive. Here, we aimed to map the interactome of this nuclear receptor and the G56R pathogenic variant through proximity labelling followed by mass spectrometry. We used the TurboID "T2A split/link design" to identify the proximity interactome of wild-type NR2E3 and mutant G56R-NR2E3 in ARPE-19 cells. Several protein complexes involved in retinal development, including the AP-1 complex, were enriched. The results suggest similar interactomes for NR2E3 and G56R-NR2E3 in ARPE-19. However, BCOR was exclusively enriched in the G56R-NR2E3 network, strengthening the connection with the IRD phenotype. In conclusion, our approach has mapped the proximity interactome of NR2E3 and G56R-NR2E3, revealing interaction with protein complexes involved in (retinal) development.
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