Death-receptor activation halts clathrin-dependent endocytosis.
Endocytosis is crucial for various aspects of cell homeostasis. Here, we show that proapoptotic death receptors (DRs) trigger selective destruction of the clathrin-dependent endocytosis machinery. DR stimulation induced rapid, caspase-mediated cleavage of key clathrin-pathway components, halting cellular uptake of the classic cargo protein transferrin. DR-proximal initiator caspases cleaved the clathrin adaptor subunit AP2alpha between functionally distinct domains, whereas effector caspases processed clathrin's heavy chain. DR5 underwent ligand-induced, clathrin-mediated endocytosis, suggesting that internalization of DR signaling complexes facilitates clathrin-pathway targeting by caspases. An endocytosis-blocking, temperature-sensitive dynamin-1 mutant attenuated DR internalization, enhanced caspase stimulation downstream of DRs, and increased apoptosis. Thus, DR-triggered caspase activity disrupts clathrin-dependent endocytosis, leading to amplification of programmed cell death.
Austin
Cary D
CD
*Departments of Research Administration, Molecular Oncology, and Protein Chemistry, Genentech, Inc., South San Francisco, CA 94080; and.
Lawrence
David A
DA
*Departments of Research Administration, Molecular Oncology, and Protein Chemistry, Genentech, Inc., South San Francisco, CA 94080; and.
Peden
Andrew A
AA
*Departments of Research Administration, Molecular Oncology, and Protein Chemistry, Genentech, Inc., South San Francisco, CA 94080; and.
Varfolomeev
Eugene E
EE
*Departments of Research Administration, Molecular Oncology, and Protein Chemistry, Genentech, Inc., South San Francisco, CA 94080; and.
Totpal
Klara
K
*Departments of Research Administration, Molecular Oncology, and Protein Chemistry, Genentech, Inc., South San Francisco, CA 94080; and.
De Mazière
Ann M
AM
Cell Microscopy Center, Department of Cell Biology and Institute for Biomembranes, University Medical Center Utrecht, 3584CX, Utrecht, The Netherlands.
Klumperman
Judith
J
Cell Microscopy Center, Department of Cell Biology and Institute for Biomembranes, University Medical Center Utrecht, 3584CX, Utrecht, The Netherlands.
Arnott
David
D
*Departments of Research Administration, Molecular Oncology, and Protein Chemistry, Genentech, Inc., South San Francisco, CA 94080; and.
Pham
Victoria
V
*Departments of Research Administration, Molecular Oncology, and Protein Chemistry, Genentech, Inc., South San Francisco, CA 94080; and.
Scheller
Richard H
RH
*Departments of Research Administration, Molecular Oncology, and Protein Chemistry, Genentech, Inc., South San Francisco, CA 94080; and aa@gene.com scheller@gene.com.
Ashkenazi
Avi
A
*Departments of Research Administration, Molecular Oncology, and Protein Chemistry, Genentech, Inc., South San Francisco, CA 94080; and aa@gene.com scheller@gene.com.
D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000494:pharmacology; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D002966:Clathrin / Q000378:metabolism*; D004705:Endocytosis / Q000187:drug effects; D004789:Enzyme Activation; D006801:Humans; D008562:Membrane Glycoproteins / Q000494:pharmacology; D016253:Microscopy, Immunoelectron; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism*; D053221:TNF-Related Apoptosis-Inducing Ligand; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D051017:Apoptosis Regulatory Proteins; D002966:Clathrin; D008562:Membrane Glycoproteins; D018124:Receptors, Tumor Necrosis Factor; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D014409:Tumor Necrosis Factor-alpha; D020169:Caspases