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The Plant Unique ESCRT Component FREE1 Regulates Autophagosome Closure

ABSTRACT: The energy sensor AMP-activated protein kinase (AMPK) can activate autophagy when cellular energy production becomes compromised. However, the degree to which nutrient sensing impinges on the autophagosome closure remains unknown. Here, we provide the mechanism underlying a plant unique protein FREE1, upon autophagy-induced SnRK1α1-mediated phosphorylation, functions as a linkage between ATG conjugation system and ESCRT machinery to regulate the autophagosome closure upon nutrient deprivation. Using high-resolution microscopy, 3D-electron tomography, and protease protection assay, we showed that unclosed autophagosomes accumulated in free1 mutants. Proteomic, cellular and biochemical analysis revealed the mechanistic connection between FREE1 and the ATG conjugation system/ESCRT-III complex in regulating autophagosome closure. Mass spectrometry analysis showed that the evolutionary conserved plant energy sensor SnRK1α1 phosphorylates FREE1 and recruits it to the autophagosomes to promote closure. Mutagenesis of the phosphorylation site on FREE1 caused the autophagosome closure failure. Our findings unveil how cellular energy sensing pathways regulate autophagosome closure to maintain cellular homeostasis.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Whole Body

SUBMITTER: Lei Feng

LAB HEAD: Lei Feng

PROVIDER: PXD040435 | Pride | 2023-03-30

REPOSITORIES: Pride

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			Action	DRS
	FREE1_rep1_fraction1_20200716.raw	Raw
	FREE1_rep1_fraction2_20200716.raw	Raw
	FREE1_rep1_fraction3_20200716.raw	Raw
	FREE1_rep1_fraction4_20200716.raw	Raw
	FREE1_rep1_fraction5_20200716.raw	Raw

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