Mechanical Allostery: Evidence for a Force Requirement in the Proteolytic Activation of Notch

Wendy R. Gordon; Brandon Zimmerman; Li He; Laura J. Miles; Jiuhong Huang; Kittichoat Tiyanont; Debbie G. McArthur; Jon C. Aster; Norbert Perrimon; Joseph J. Loparo; Stephen C. Blacklow

doi:10.1016/j.devcel.2015.05.004

Mechanical Allostery: Evidence for a Force Requirement in the Proteolytic Activation of Notch

Wendy R. Gordon, Brandon Zimmerman, Li He, Laura J. Miles, Jiuhong Huang, Kittichoat Tiyanont, Debbie G. McArthur, Jon C. Aster, Norbert Perrimon, Joseph J. Loparo, Stephen C. Blacklow

Chemical and Structural Biology (TMED)

Research output: Contribution to journal › Article › peer-review

249 Scopus citations

Abstract

Ligands stimulate Notch receptors by inducing regulated intramembrane proteolysis (RIP) to produce a transcriptional effector. Notch activation requires unmasking of a metalloprotease cleavage site remote from the site of ligand binding, raising the question of how proteolytic sensitivity is achieved. Here, we show that application of physiologically relevant forces to the Notch1 regulatory switch results in sensitivity to metalloprotease cleavage, and bound ligands induce Notch signal transduction in cells only in the presence of applied mechanical force. Synthetic receptor-ligand systems that remove the native ligand-receptor interaction also activate Notch by inducing proteolysis of the regulatory switch. Together, these studies show that mechanical force exerted by signal-sending cells is required for ligand-induced Notch activation and establish that force-induced proteolysis can act as a mechanism of cellular mechanotransduction.

Original language	English (US)
Pages (from-to)	729-736
Number of pages	8
Journal	Developmental Cell
Volume	33
Issue number	6
DOIs	https://doi.org/10.1016/j.devcel.2015.05.004
State	Published - Jun 22 2015

Bibliographical note

Funding Information:
We thank Benedikt Bauer and Anthony Nguyen for help in optimizing single molecule experiments and Adam17 expression, Scott Ficarro for mass spectrometry, and Kelly Arnett and Brian McMillan for helpful discussions. W.R.G. was supported by an American Heart Association (AHA) SDG grant. L.H. was supported by a Damon Runyon Postdoctoral Fellowship. This work was supported by NIH grants P01 CA119070 (J.J.L., J.C.A., and S.C.B.) and R01 CA092433 (J.C.A. and S.C.B.), the Howard Hughes Medical Institute (HHMI), and the NIH (N.P.).

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© 2015 Elsevier Inc.

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abstract = "Ligands stimulate Notch receptors by inducing regulated intramembrane proteolysis (RIP) to produce a transcriptional effector. Notch activation requires unmasking of a metalloprotease cleavage site remote from the site of ligand binding, raising the question of how proteolytic sensitivity is achieved. Here, we show that application of physiologically relevant forces to the Notch1 regulatory switch results in sensitivity to metalloprotease cleavage, and bound ligands induce Notch signal transduction in cells only in the presence of applied mechanical force. Synthetic receptor-ligand systems that remove the native ligand-receptor interaction also activate Notch by inducing proteolysis of the regulatory switch. Together, these studies show that mechanical force exerted by signal-sending cells is required for ligand-induced Notch activation and establish that force-induced proteolysis can act as a mechanism of cellular mechanotransduction.",

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Mechanical Allostery: Evidence for a Force Requirement in the Proteolytic Activation of Notch

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