Cleavage of Notch by furin is required to generate a mature, cell surface heterodimeric receptor that can be proteolytically activated to release its intracellular domain, which functions in signal transduction. Current models propose that ligand binding to heterodimeric Notch (hNotch) induces a disintegrin and metalloprotease (ADAM) proteolytic release of the Notch extracellular domain (NECD), which is subsequently shed and/or endocytosed by DSL ligand cells. We provide evidence for NECD release and internalization by DSL ligand cells, which, surprisingly, did not require ADAM activity. However, losses in either hNotch formation or ligand endocytosis significantly decreased NECD transfer to DSL ligand cells, as well as signaling in Notch cells. Because endocytosis-defective ligands bind hNotch, but do not dissociate it, additional forces beyond those produced through ligand binding must function to disrupt the intramolecular interactions that keep hNotch intact and inactive. Based on our findings, we propose that mechanical forces generated during DSL ligand endocytosis function to physically dissociate hNotch, and that dissociation is a necessary step in Notch activation.
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12 February 2007
Article|
February 12 2007
DSL ligand endocytosis physically dissociates Notch1 heterodimers before activating proteolysis can occur
James T. Nichols,
James T. Nichols
1Department of Biological Chemistry, David Geffen School of Medicine
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Alison Miyamoto,
Alison Miyamoto
1Department of Biological Chemistry, David Geffen School of Medicine
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Samantha L. Olsen,
Samantha L. Olsen
1Department of Biological Chemistry, David Geffen School of Medicine
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Brendan D'Souza,
Brendan D'Souza
1Department of Biological Chemistry, David Geffen School of Medicine
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Christine Yao,
Christine Yao
1Department of Biological Chemistry, David Geffen School of Medicine
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Gerry Weinmaster
Gerry Weinmaster
1Department of Biological Chemistry, David Geffen School of Medicine
2Molecular Biology Institute,
3Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA 90095
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James T. Nichols
1Department of Biological Chemistry, David Geffen School of Medicine
Alison Miyamoto
1Department of Biological Chemistry, David Geffen School of Medicine
Samantha L. Olsen
1Department of Biological Chemistry, David Geffen School of Medicine
Brendan D'Souza
1Department of Biological Chemistry, David Geffen School of Medicine
Christine Yao
1Department of Biological Chemistry, David Geffen School of Medicine
Gerry Weinmaster
1Department of Biological Chemistry, David Geffen School of Medicine
2Molecular Biology Institute,
3Jonsson Comprehensive Cancer Center, University of California, Los Angeles, Los Angeles, CA 90095
Correspondence to Gerry Weinmaster: [email protected]
Abbreviations used in this paper: ADAM, a disintegrin and metalloprotease; CSL, CBF1, SuH, LAG-1; DIC, differential interference contrast; DSL, Delta/Serrate/Lag-2; hNotch, heterodimeric Notch; NECD, Notch extracellular domain; NICD, Notch intracellular domain; RIP, regulated intramembrane proteolysis.
Received:
September 05 2006
Accepted:
January 12 2007
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2007
J Cell Biol (2007) 176 (4): 445–458.
Article history
Received:
September 05 2006
Accepted:
January 12 2007
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Endocytosis pulls Notch apart
Citation
James T. Nichols, Alison Miyamoto, Samantha L. Olsen, Brendan D'Souza, Christine Yao, Gerry Weinmaster; DSL ligand endocytosis physically dissociates Notch1 heterodimers before activating proteolysis can occur . J Cell Biol 12 February 2007; 176 (4): 445–458. doi: https://doi.org/10.1083/jcb.200609014
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