title: Mechanical Allostery: Evidence for a Force Requirement in the Proteolytic Activation of Notch.
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.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Wendy R
authors.initials: WR
authors.lastname: Gordon
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Dana Farber Cancer Institute, Boston, MA 02215, USA.
authors.forename: Brandon
authors.initials: B
authors.lastname: Zimmerman
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Li
authors.initials: L
authors.lastname: He
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Dana Farber Cancer Institute, Boston, MA 02215, USA.
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Miles
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA 02115, USA; School of Life Science and Technology, Tongji University, Shanghai 200092, China.
authors.forename: Jiuhong
authors.initials: J
authors.lastname: Huang
authors.affiliation: Dana Farber Cancer Institute, Boston, MA 02215, USA.
authors.forename: Kittichoat
authors.initials: K
authors.lastname: Tiyanont
authors.affiliation: Dana Farber Cancer Institute, Boston, MA 02215, USA.
authors.forename: Debbie G
authors.initials: DG
authors.lastname: McArthur
authors.affiliation: Brigham and Women's Hospital, Boston, MA 02115, USA.
authors.forename: Jon C
authors.initials: JC
authors.lastname: Aster
authors.affiliation: Department of Genetics, Harvard Medical School, Boston, MA 02115, USA; Howard Hughes Medical Institute, Boston, MA 02115, USA.
authors.forename: Norbert
authors.initials: N
authors.lastname: Perrimon
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA. Electronic address: joseph_loparo@hms.harvard.edu.
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Loparo
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA; Dana Farber Cancer Institute, Boston, MA 02215, USA; Brigham and Women's Hospital, Boston, MA 02115, USA. Electronic address: stephen_blacklow@hms.harvard.edu.
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Blacklow
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism; D000072198:ADAM17 Protein; D000494:Allosteric Regulation; D000818:Animals; D058045:Artificial Cells; D001696:Biomechanical Phenomena; D002460:Cell Line; D004705:Endocytosis; D057809:HEK293 Cells; D006801:Humans; D008024:Ligands; D040542:Mechanotransduction, Cellular; D008954:Models, Biological; D059748:Proteolysis; D051880:Receptors, Notch / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D008024:Ligands; D051880:Receptors, Notch; D051722:ADAM Proteins; D000072198:ADAM17 Protein