RAB24 facilitates clearance of autophagic compartments during basal conditions

Autophagy
Päivi Ylä-AnttilaEeva-Liisa Eskelinen

Abstract

RAB24 belongs to a family of small GTPases and has been implicated to function in autophagy. Here we confirm the intracellular localization of RAB24 to autophagic vacuoles with immuno electron microscopy and cell fractionation, and show that prenylation and guanine nucleotide binding are necessary for the targeting of RAB24 to autophagic compartments. Further, we show that RAB24 plays a role in the maturation and/or clearance of autophagic compartments under nutrient-rich conditions, but not during short amino acid starvation. Quantitative electron microscopy shows an increase in the numbers of late autophagic compartments in cells silenced for RAB24, and mRFP-GFP-LC3 probe and autophagy flux experiments indicate that this is due to a hindrance in their clearance. Formation of autophagosomes is shown to be unaffected by RAB24-silencing with siRNA. A defect in aggregate clearance in the absence of RAB24 is also shown in cells forming polyglutamine aggregates. This study places RAB24 function in the termination of the autophagic process under nutrient-rich conditions.

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Citations

Mar 18, 2016·Autophagy·Navodita Jain, Subramaniam Ganesh
Dec 23, 2016·Small GTPases·Szabolcs TakátsGábor Juhász
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Methods Mentioned

BETA
GTPases
GTPase
confocal microscopy
transmission electron microscopy
electron microscopy
transfection
dot
filter
filter trap
fluorescence microscopy

Software Mentioned

CellProfiler
Colocalization finder Plugin
RunImageJ
IdentifyPrimaryObjects
ImageJ
Image
IdentifySecondaryObjects
Pro Plus
ImageJ finder Plugin

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