CADASIL mutations enhance spontaneous multimerization of NOTCH3

Author： Opherk Christian Duering Marco Peters Nils Karpinska Anna Rosner Stefanie Schneider Elisabeth Bader Benedikt Giese Armin Dichgans Martin

Publisher： Oxford University Press

ISSN： 1460-2083

Source： Human Molecular Genetics, Vol.18, Iss.15, 2009-08, pp. : 2761-2767

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Abstract

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common monogenic cause of stroke and vascular dementia. Disease-causing mutations invariably affect cysteine residues within epidermal growth factor-like repeat domains in the extracellular domain of the NOTCH3 receptor (N3^ECD). The biochemical and histopathological hallmark of CADASIL is the accumulation of N3^ECD at the cell surface of vascular smooth muscle cells which degenerate over the course of the disease. The molecular mechanisms leading to N3^ECD accumulation remain unknown. Here we show that both wild-type and CADASIL-mutated N3^ECD spontaneously form oligomers and higher order multimers in vitro and that multimerization is mediated by disulfide bonds. Using single-molecule analysis techniques (scanning for intensely fluorescent targets), we demonstrate that CADASIL-associated mutations significantly enhance multimerization compared with wild-type. Taken together, our results for the first time provide experimental evidence for N3 self-association and strongly argue for a neomorphic effect of CADASIL mutations in disease pathogenesis.