The small G protein Arl5 contributes to endosome-to-Golgi traffic by aiding the recruitment of the GARP complex to the Golgi

Biology Open
Cláudia Rosa-FerreiraSean Munro

Abstract

The small G proteins of the Arf family play critical roles in membrane trafficking and cytoskeleton organization. However, the function of some members of the family remains poorly understood including Arl5 which is widely conserved in eukaryotes. Humans have two closely related Arl5 paralogues (Arl5a and Arl5b), and both Arl5a and Arl5b localize to the trans-Golgi with Arl5b being involved in retrograde traffic from endosomes to the Golgi apparatus. To investigate the function of Arl5, we have used Drosophila melanogaster as a model system. We find that the single Arl5 orthologue in Drosophila also localizes to the trans-Golgi, but flies lacking the Arl5 gene are viable and fertile. By using both liposome and column based affinity chromatography methods we find that Arl5 interacts with the Golgi-associated retrograde protein (GARP) complex that acts in the tethering of vesicles moving from endosomes to the trans-Golgi network (TGN). In Drosophila tissues the GARP complex is partially displaced from the Golgi when Arl5 is absent, and the late endosomal compartment is enlarged. In addition, in HeLa cells GARP also becomes cytosolic upon depletion of Arl5b. These phenotypes are consistent with a role in endosome-to-Golgi traffic,...Continue Reading

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Citations

Mar 15, 2016·Trends in Cell Biology·Alison K Gillingham, Sean Munro
Aug 3, 2016·Biology Open·Karolina FáriRita Sinka
May 16, 2019·Molecular Biology of the Cell·Elizabeth SztulRichard A Kahn
Apr 24, 2020·Proceedings of the National Academy of Sciences of the United States of America·María Otilia DelgadilloEnrique Rojo
Feb 16, 2021·Frontiers in Cell and Developmental Biology·Kana Fujibayashi, Joji Mima
Nov 2, 2016·Journal of Cell Science·Cinzia Progida, Oddmund Bakke
Jul 12, 2021·Genome Biology and Evolution·Romana VargováMarek Eliáš

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Methods Mentioned

BETA
PCR
transgenic
transfection
FCS
affinity purifications
affinity purification

Software Mentioned

ImageJ
Fiji

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