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Article
2024
tsCRISPR based identification of Rab proteins required for the recycling of Drosophila TRPL ion channel
tsCRISPR based identification of Rab proteins required for the recycling of Drosophila TRPL ion channel
Abstract (English)
In polarized cells, the precise regulation of protein transport to and from the
plasma membrane is crucial to maintain cellular function. Dysregulation of
intracellular protein transport in neurons can lead to neurodegenerative
diseases such as Retinitis Pigmentosa, Alzheimer’s and Parkinson’s disease.
Here we used the light-dependent transport of the TRPL (transient receptor
potential-like) ion channel in Drosophila photoreceptor cells to study the role of
Rab proteins in TRPL recycling. TRPL is located in the rhabdomeric membrane of
dark-adapted flies, but it is transported out of the rhabdomere upon light
exposure and localizes at the Endoplasmatic Reticulum within 12 h. Upon
subsequent dark adaptation, TRPL is recycled back to the rhabdomeric
membrane within 90 min. To screen for Rab proteins involved in TRPL
recycling, we established a tissue specific (ts) CRISPR/Cas9-mediated knock-
out of individual Rab genes in Drosophila photoreceptors and assessed TRPL
localization using an eGFP tagged TRPL protein in the intact eyes of these
mutants. We observed severe TRPL recycling defects in the knockouts of
Rab3, Rab4, Rab7, Rab32, and RabX2. Using immunohistochemistry, we
further showed that Rab3 and RabX2 each play a significant role in TRPL
recycling and also influence TRPL transport. We localized Rab3 to the late
endosome in Drosophila photoreceptors and observed disruption of TRPL
transport to the ER in Rab3 knock-out mutants. TRPL transport from the ER
to the rhabdomere ensues from the trans-Golgi where RabX2 is located. We
observed accumulated TRPL at the trans-Golgi in RabX2 knock-out mutants. In
summary, our study reveals the requirement of specific Rab proteins for different
steps of TRPL transport in photoreceptor cells and provides evidence for a unique
retrograde recycling pathway of TRPL from the ER via the trans-Golgi
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Frontiers in cell and developmental biology, 12 (2024), 1444953.
https://doi.org/10.3389/fcell.2024.1444953.
ISSN: 2296-634X
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@article{Zeger2024,
url = {https://hohpublica.uni-hohenheim.de/handle/123456789/16716},
doi = {10.3389/fcell.2024.1444953},
author = {Zeger, Matthias and Stanisławczyk, Lena Sarah and Bulić,Marija et al.},
title = {tsCRISPR based identification of Rab proteins required for the recycling of Drosophila TRPL ion channel},
journal = {Frontiers in cell and developmental biology},
year = {2024},
volume = {12},
}