Cholesterol and ORP1L-mediated ER contact sites control autophagosome transport and fusion with the endocytic pathway.
Autophagy is the main homeostatic pathway guiding cytosolic materials for degradation by the lysosome. Maturation of autophagosomes requires their transport towards the perinuclear region of the cell, with key factors underlying both processes still poorly understood. Here we show that transport and positioning of late autophagosomes depends on cholesterol by way of the cholesterol-sensing Rab7 effector ORP1L. ORP1L localizes to late autophagosomes and-under low-cholesterol conditions-contacts the ER protein VAP-A, forming ER-autophagosome contact sites, which prevent minus-end transport by the Rab7-RILP-dynein complex. ORP1L-mediated contact sites also inhibit localization of PLEKHM1 to Rab7. PLEKHM1, together with RILP, then recruits the homotypic fusion and vacuole protein-sorting (HOPS) complex for fusion of autophagosomes with late endosomes and lysosomes. Thus, ORP1L, via its liganding by lipids and the formation of contacts between autophagic vacuoles and the ER, governs the last steps in autophagy that lead to the lysosomal degradation of cytosolic material.
Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
Ruud H
RH
Wijdeven
Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
Hans
H
Janssen
Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
Leila
L
Nahidiazar
Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.|Department of Chemical Immunology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Lennert
L
Janssen
Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
Kees
K
Jalink
Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.|Department of Chemical Immunology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Ilana
I
Berlin
Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.|Department of Chemical Immunology, Leiden University Medical Center, 2300 RC Leiden, The Netherlands.
Jacques
J
Neefjes
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000071182:Autophagosomes / Q000378:metabolism* / Q000648:ultrastructure; D000071183:Autophagy-Related Proteins; D002784:Cholesterol / Q000378:metabolism*; D004705:Endocytosis; D004721:Endoplasmic Reticulum / Q000378:metabolism* / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D008561:Membrane Fusion; D008562:Membrane Glycoproteins / Q000378:metabolism; D008954:Models, Biological; D011485:Protein Binding; D021381:Protein Transport; D011987:Receptors, Steroid / Q000378:metabolism*; D014617:Vacuoles / Q000378:metabolism / Q000648:ultrastructure; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D000098825:Oxysterol Binding Proteins
D048868:Adaptor Proteins, Signal Transducing; D000071183:Autophagy-Related Proteins; D008562:Membrane Glycoproteins; C476215:PLEKHM1 protein, human; C423317:RILP protein, human; D011987:Receptors, Steroid; D000098825:Oxysterol Binding Proteins; D002784:Cholesterol; D020691:rab GTP-Binding Proteins