Unraveling the impact of VHL exon 2 mutations in erythrocytosis or von Hippel-Lindau disease identified RNA-binding proteins involved in VHL splicing.

This study explores the impact of mutations in the exon 2 of the von Hippel-Lindau (VHL) gene, associated with erythrocytosis or von Hippel-Lindau disease. We analyzed 15 missense and synonymous genetic variants to assess their effects on splicing and VHL protein stability. Using in silico predictions and functional assays, we found that some specific mutations impact splicing and reduce protein stability, allowing their clinical classification as pathogenic. This study revealed exonic-splicing regulatory regions. Notably, by performing RNA-protein pull-down, we identified two RNA-binding proteins, hnRNPF and hnRNPAB, as key regulators of VHL splicing. Our findings reveal the limitations of current splicing-prediction tools in recognizing exonic-splicing enhancer (ESE) or silencer (ESS) sequences and suggest that mutations can differentially affect disease phenotypes by influencing both splicing and protein stability. These insights enhance our understanding of the molecular mechanisms underlying VHL-associated disorders and expand the landscape of regulatory elements and protein factors involved in VHL splicing regulation.