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

Dev Cell. 2015 Jun 22;33(6):729-36. doi: 10.1016/j.devcel.2015.05.004. Epub 2015 Jun 4.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins / metabolism
  • ADAM17 Protein
  • Allosteric Regulation
  • Animals
  • Artificial Cells
  • Biomechanical Phenomena
  • Cell Line
  • Endocytosis
  • HEK293 Cells
  • Humans
  • Ligands
  • Mechanotransduction, Cellular
  • Models, Biological
  • Proteolysis
  • Receptors, Notch / chemistry
  • Receptors, Notch / metabolism*
  • Signal Transduction

Substances

  • Ligands
  • Receptors, Notch
  • ADAM Proteins
  • ADAM17 Protein