Purchase this article with an account.
Rob W. Collin, Konstantinos Nikopoulos, Christian Gilissen, F Nienke Boonstra, Hiroyuki Kondo, Carmel Toomes, Wolfgang Berger, C Erik van Nouhuys, Alexander Hoischen, Frans P. Cremers; Combined Exome Sequencing And Linkage Analysis Reveals A Dominant-negative ZNF408 Mutation Causing Familial Exudative Vitreoretinopathy. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2257.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To identify the genetic defect underlying autosomal dominant familial exudative vitreoretinopathy (FEVR) in a large Dutch pedigree.
Affected family members were genotyped using 6k SNP arrays, followed by linkage analysis. Two distantly related patients were analyzed by exome sequencing. Candidate variants were validated and segregation analysis was performed in the complete family. Upon identification of a causative missense mutation in ZNF408, this gene was screened in a cohort of 132 additional FEVR probands. Subsequently, cDNA constructs expressing wild-type and mutant ZNF408 proteins were generated for immunolocalization studies in COS-1 cells. The tissue distribution of ZNF408 was studied via real-time quantitative RT-PCR analysis.
Combining exome sequencing, linkage analysis and segregation analysis revealed a single probable disease-causing missense mutation in ZNF408 (c.1363C>T; p.His455Tyr), which was also detected in an additional Dutch FEVR family. Sequence analysis of ZNF408 in 132 additional FEVR patients revealed a second missense variant (p.Ser126Asn). Immunolocalisation analysis revealed that the wild-type, and the p.Ser126Asn mutant protein show a complete nuclear localization, whereas the p.His455Tyr mutant protein was localized almost exclusively in the cytoplasm. Moreover, in a co-transfection assay, the p.His455Tyr mutant protein retains the wild-type ZNF408 protein in the cytoplasm, suggesting a dominant-negative effect of the p.His455Tyr mutation. ZNF408 was found to be abundantly expressed in the retina.
Together, our data strongly suggest that ZNF408 is implicated in adFEVR, thereby revealing a transcription factor that may act in the norrin/β-catenin signaling pathway.
This PDF is available to Subscribers Only