Arginine/Serine-rich Protein Interaction Domain-dependent Modulation of a Tau Exon 10 Splicing Enhancer

Tau exon 10 splicing is altered by autosomal dominant mutations that cause frontotemporal dementia with parkinsonism chromosome 17-type and by unknown mechanisms in other related neurodegenerative disorders. Identifying cis- and trans-regulators of tau exon 10 splicing is therefore crucial for understanding disease mechanisms. We previously identified several splicing enhancers and silencers within exon 10 and intron 10. Here, we show that splicing factors SF2/ASF, Tra2β, and a 50-kDa nuclear protein bind in vitro to the polypurine enhancer at the 5′ end of exon 10. Disease splicing mutations N279K and Δ280K disrupt the enhancer and alter associations with these factors. N279K targets robustly bind Tra2β compared with the normal enhancer, which may explain why N279K enhances exon 10 splicing in vivo. In contrast, factor associations with Δ280K targets are nearly undetectable, explaining why Δ280K almost abolishes exon 10 splicing in vivo. Small interfering RNA-mediated suppression of endogenous SF2/ASF and Tra2β significantly reduces exon 10 splicing. Exogenous SF2/ASF dramatically enhances normal exon 10 splicing and efficiently rescues the Δ280K splicing defect. Domain deletion analyses show that the C-terminal RS domains of SF2/ASF and Tra2β are required for normal exon 10 splicing in vivo. In contrast to Tra2β, the SF2/ASF RS domain remains essential in the presence of a strengthened enhancer or when either weak splice site is strengthened. The data suggest that SF2/ASF has both essential and regulatory roles, whereas Tra2β has a supporting role in exon 10 splicing.

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