TY - JOUR
T1 - Endosomal LC3C-pathway selectively targets plasma membrane cargo for autophagic degradation
AU - Coelho, Paula P.
AU - Hesketh, Geoffrey G.
AU - Pedersen, Annika
AU - Kuzmin, Elena
AU - Fortier, Anne Marie N.
AU - Bell, Emily S.
AU - Ratcliffe, Colin D.H.
AU - Gingras, Anne Claude
AU - Park, Morag
N1 - Funding Information:
We thank members of the Park laboratory for their helpful comments on the manuscript; and Genentech Inc. for HGF. Images were collected for this manuscript was performed in the McGill University Advanced BioImaging Facility (ABIF), RRID:SCR_017697. Proteomics work was performed at the Network Biology Collaborative Centre at the Lunenfeld-Tanenbaum Research Institute, a facility supported by Canada Foundation for Innovation funding, by the Ontario Government, and by Genome Canada and Ontario Genomics (OGI-139). The flow cytometry work was performed in the Flow Cytometry Core Facility of the Life Science Complex and supported by funding from the Canadian Foundation for Innovation. This research was supported by doctoral studentships from the Fonds de Recherche du Québec – Santé to P.P.C. and C.D.H.R. and the Rosalind Goodman Commemorative Scholarship to P.P.C. and C.D.H.R.; the United States Department of Defense (BC100836) to E.S.B.; the GCRC Recruitment Scholarship to A.P.; and Foundation operating grants to N.So. (148423), M.P. (242529) and A.C.G. (143301) from the Canadian Institutes of Health Research. M.P. holds the Diane and Sal Guerrera Chair in Cancer Genetics. And A.C.G. is the Canadian Research Chair in Functional Proteomics.
Funding Information:
We thank members of the Park laboratory for their helpful comments on the manuscript; and Genentech Inc. for HGF. Images were collected for this manuscript was performed in the McGill University Advanced BioImaging Facility (ABIF), RRID:SCR_017697. Proteomics work was performed at the Network Biology Collaborative Centre at the Lunenfeld-Tanenbaum Research Institute, a facility supported by Canada Foundation for Innovation funding, by the Ontario Government, and by Genome Canada and Ontario Genomics (OGI-139). The flow cytometry work was performed in the Flow Cytometry Core Facility of the Life Science Complex and supported by funding from the Canadian Foundation for Innovation. This research was supported by doctoral studentships from the Fonds de Recherche du Québec – Santé to P.P.C. and C.D.H.R. and the Rosalind Goodman Commemorative Scholarship to P.P.C. and C.D.H.R.; the United States Department of Defense (BC100836) to E.S.B.; the GCRC Recruitment Scholarship to A.P.; and Foundation operating grants to N.So. (148423), M.P. (242529) and A.C.G. (143301) from the Canadian Institutes of Health Research. M.P. holds the Diane and Sal Guerrera Chair in Cancer Genetics. And A.C.G. is the Canadian Research Chair in Functional Proteomics.
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - Autophagy selectively targets cargo for degradation, yet mechanistic understanding remains incomplete. The ATG8-family plays key roles in autophagic cargo recruitment. Here by mapping the proximal interactome of ATG8-paralogs, LC3B and LC3C, we uncover a LC3C-Endocytic-Associated-Pathway (LEAP) that selectively recruits plasma-membrane (PM) cargo to autophagosomes. We show that LC3C localizes to peripheral endosomes and engages proteins that traffic between PM, endosomes and autophagosomes, including the SNARE-VAMP3 and ATG9, a transmembrane protein essential for autophagy. We establish that endocytic LC3C binds cargo internalized from the PM, including the Met receptor tyrosine kinase and transferrin receptor, and is necessary for their recruitment into ATG9 vesicles targeted to sites of autophagosome initiation. Structure-function analysis identified that LC3C-endocytic localization and engagement with PM-cargo requires the extended carboxy-tail unique to LC3C, the TBK1 kinase, and TBK1-phosphosites on LC3C. These findings identify LEAP as an unexpected LC3C-dependent pathway, providing new understanding of selective coupling of PM signalling with autophagic degradation.
AB - Autophagy selectively targets cargo for degradation, yet mechanistic understanding remains incomplete. The ATG8-family plays key roles in autophagic cargo recruitment. Here by mapping the proximal interactome of ATG8-paralogs, LC3B and LC3C, we uncover a LC3C-Endocytic-Associated-Pathway (LEAP) that selectively recruits plasma-membrane (PM) cargo to autophagosomes. We show that LC3C localizes to peripheral endosomes and engages proteins that traffic between PM, endosomes and autophagosomes, including the SNARE-VAMP3 and ATG9, a transmembrane protein essential for autophagy. We establish that endocytic LC3C binds cargo internalized from the PM, including the Met receptor tyrosine kinase and transferrin receptor, and is necessary for their recruitment into ATG9 vesicles targeted to sites of autophagosome initiation. Structure-function analysis identified that LC3C-endocytic localization and engagement with PM-cargo requires the extended carboxy-tail unique to LC3C, the TBK1 kinase, and TBK1-phosphosites on LC3C. These findings identify LEAP as an unexpected LC3C-dependent pathway, providing new understanding of selective coupling of PM signalling with autophagic degradation.
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U2 - 10.1038/s41467-022-31465-3
DO - 10.1038/s41467-022-31465-3
M3 - Article
C2 - 35780247
AN - SCOPUS:85133352801
SN - 2041-1723
VL - 13
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 3812
ER -