Unusual splice site mutations disrupt FANCA exon 8 definition

作者：Mattioli Chiara; Pianigiani Giulia; De Rocco Daniela; Bianco Anna Monica Rosaria; Cappelli Enrico; Savoia Anna; Pagani Franco^*

来源：Biochimica et Biophysica Acta-Molecular Basis of Disease, 2014, 1842(7): 1052-1058.

DOI：10.1016/j.bbadis.2014.03.014

• Nonsense codon
• Fanconi anemia
• Nonsense mediated mRNA decay
• U1 small nuclear ribonucleoprotein
• RNA splice sites
• RNA splicing

The pathological role of mutations that affect not conserved splicing regulatory sequences can be difficult to determine. In a patient with Fanconi anemia, we identified two unpredictable splicing mutations that act on either sides of FANCA exon 8. In patients-derived cells and in minigene splicing assay, we showed that both an apparently benign intronic c.710-5T>C transition and the nonsense c.790C>T substitution induce almost complete exon 8 skipping. Site-directed mutagenesis experiments indicated that the c.710-5T>C transition affects a polypyrimidine tract where most of the thymidines cannot be compensated by cytidines. The c.790C>T mutation located in position 3 relative to the donor site induce exon 8 skipping in an NMD-independent manner and complementation experiments with modified U1 snRNAs showed that U1 snRNP is only partially involved in the splicing defect. Our results highlight the importance of performing splicing functional assay for correct identification of disease-causing mechanism of genomic variants and provide mechanistic insights on how these two FANCA mutations affect exon 8 definition.

• 出版日期2014-7