title: CADASIL-causing mutations do not alter Notch3 receptor processing and activation.
abstract: CADASIL is associated with mutations in the Notch3 gene but the causal mechanisms of the disorder remain unclear. We studied effects of widely established mutations on Notch3 receptor processing and ligand-mediated activation in stable lines of HEK293 and SH-SY5Y cells expressing either human wild-type or mutant Notch3 receptor. None of the four mutations (R90C, R133C, C185R and R449C) affected quantities of the full-length, amino-terminal or carboxyl-terminal fragments and did not impair intracellular trafficking in both cell types. The Jagged 1, Jagged 2 and Delta ligand-mediated S2 site cleavage and signal transduction were also observed to be similar in both wild-type and mutants, which exhibited similar rates of degradation of full-length, amino-terminal and carboxyl-terminal fragments. Our results suggest that the arteriopathy in CADASIL is caused by other mechanisms not necessarily involving Notch3 processing and activation.
authors.affiliation: Institute for Ageing and Health, University of Newcastle, Newcastle upon Tyne, UK.
authors.forename: Wee-Chuang
authors.initials: WC
authors.lastname: Low
authors.forename: Yo
authors.initials: Y
authors.lastname: Santa
authors.forename: Keikichi
authors.initials: K
authors.lastname: Takahashi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tabira
authors.forename: Raj N
authors.initials: RN
authors.lastname: Kalaria
mesh_terms: D015153:Blotting, Western / Q000379:methods; D046589:CADASIL / Q000235:genetics*; D002460:Cell Line; D018920:Coculture Techniques / Q000379:methods; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D008024:Ligands; D008156:Luciferases / Q000378:metabolism; D008970:Molecular Weight; D016296:Mutagenesis / Q000502:physiology; D009154:Mutation; D011485:Protein Binding / Q000235:genetics; D000071656:Receptor, Notch3; D051880:Receptors, Notch / Q000502:physiology*; D015398:Signal Transduction / Q000235:genetics*; D014162:Transfection / Q000379:methods
chemical_list: D008024:Ligands; C499374:NOTCH3 protein, human; D000071656:Receptor, Notch3; D051880:Receptors, Notch; D008156:Luciferases