TY - JOUR
T1 - The plant unique ESCRT component FREE1 regulates autophagosome closure
AU - Zeng, Yonglun
AU - Li, Baiying
AU - Huang, Shuxian
AU - Li, Hongbo
AU - Cao, Wenhan
AU - Chen, Yixuan
AU - Liu, Guoyong
AU - Li, Zhenping
AU - Yang, Chao
AU - Feng, Lei
AU - Gao, Jiayang
AU - Lo, Sze Wan
AU - Zhao, Jierui
AU - Shen, Jinbo
AU - Guo, Yan
AU - Gao, Caiji
AU - Dagdas, Yasin
AU - Jiang, Liwen
N1 - Publisher Copyright:
© 2023, The Author(s).
PY - 2023/3/30
Y1 - 2023/3/30
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85151316330&partnerID=8YFLogxK
U2 - 10.1038/s41467-023-37185-6
DO - 10.1038/s41467-023-37185-6
M3 - Journal article
C2 - 36997511
AN - SCOPUS:85151316330
SN - 2041-1723
VL - 14
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 1768
ER -
