DRAM1 requires PI(3,5)P2 generation by PIKfyve to deliver vesicles and their cargo to endolysosomes
| Main Author: | Michiel van der Vaart |
|---|---|
| Other Authors: | Adrianna Banducci-Karp, Gabriel Forn-Cuní, Philip M. M. Witt, Joost J. Willemse, Salomé Muñoz Sánchez, Rohola Hosseini, Annemarie H. Meijer |
| Format: | info publication-preprint Journal |
| Bahasa: | eng |
| Terbitan: |
, 2021
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| Subjects: | |
| Online Access: |
https://zenodo.org/record/4434243 |
Daftar Isi:
- Using high-resolution confocal microscopy in the zebrafish model, we show that mCherry-Dram1 labelled vesicles interact and fuse with early endosomes marked by PI(3)P. Following these fusion events, early endosomes mature into late endosomes in a process dependent on the conversion of PI(3)P into PI(3,5)P2 by the lipid kinase PIKfyve. Chemical inhibition of PIKfyve reduces the targeting of Dram1 to acidic endolysosomal vesicles, arresting Dram1 in multivesicular bodies, early endosomes, or non-acidified vesicles halted in their fusion with early endosomes. Zebrafish line referred to as mCherry-Dram1: Tg(bactin:mCherry-dram1) (DNA described in: Van der Vaart et al., 2014) Zebrafish line referred to as GFP-2xFYVE: TgBAC(ΔNp63:Gal4FF)la213; Tg(4xUAS:EGFP-2xFYVE)la214 (Rasmussen et al., 2015)