Characterization of a disease-associated mutation affecting a putative splicing regulatory element in intron 6b of the cystic fibrosis transmembrane conductance regulator (CFTR) gene

Valeria Faà, Federica Incani, Alessandra Meloni, Denise Corda, Maddalena Masala, A. Mario Baffico, Manuela Seia, Antonio Cao, M. Cristina Rosatelli

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Cystic fibrosis (CF) is a common recessive disorder caused by >1600 mutations in the CF transmembrane conductance regulator (CFTR) gene. About 13% of CFTR mutations are classified as "splicing mutations," but for almost 40% of these, their role in affecting the pre-mRNA splicing of the gene is not yet defined. In this work, we describe a new splicing mutation detected in three unrelated Italian CF patients. By DNA analyses and mRNA studies, we identified the c.1002-1110_1113delTAAG mutation localized in intron 6b of the CFTR gene. At themRNA level, this mutation creates an aberrant inclusion of a sequence of 101 nucleotides between exons 6b and 7. This sequence corresponds to a portion of intron 6b and resembles a cryptic exon because it is characterized by an upstream ag and a downstream gt sequence, which are most probably recognized as 5′- and 3′-splice sites by the spliceosome. Through functional analysis of this splicing defect, we show that this mutation abolishes the interaction of the splicing regulatory protein heterogeneous nuclear ribonucleoprotein A2/B1 with an intronic splicing regulatory element and creates a new recognition motif for the SRp75 splicing factor, causing activation of the cryptic exon. Our results show that the c.1002-1110_1113delTAAG mutation creates a new intronic splicing regulatory element in intron 6b of the CFTR gene exclusively recognized by SRp75.

Original languageEnglish
Pages (from-to)30024-30031
Number of pages8
JournalJournal of Biological Chemistry
Volume284
Issue number44
DOIs
Publication statusPublished - Oct 30 2009

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Cystic Fibrosis Transmembrane Conductance Regulator
Regulator Genes
Introns
Genes
Exons
RNA Splice Sites
Mutation
Heterogeneous-Nuclear Ribonucleoprotein Group A-B
Functional analysis
RNA Precursors
Cystic Fibrosis
Nucleotides
Chemical activation
Protein Splicing
Spliceosomes
Messenger RNA
Defects
DNA
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

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Characterization of a disease-associated mutation affecting a putative splicing regulatory element in intron 6b of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. / Faà, Valeria; Incani, Federica; Meloni, Alessandra; Corda, Denise; Masala, Maddalena; Baffico, A. Mario; Seia, Manuela; Cao, Antonio; Rosatelli, M. Cristina.

In: Journal of Biological Chemistry, Vol. 284, No. 44, 30.10.2009, p. 30024-30031.

Research output: Contribution to journalArticle

Faà, Valeria ; Incani, Federica ; Meloni, Alessandra ; Corda, Denise ; Masala, Maddalena ; Baffico, A. Mario ; Seia, Manuela ; Cao, Antonio ; Rosatelli, M. Cristina. / Characterization of a disease-associated mutation affecting a putative splicing regulatory element in intron 6b of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 44. pp. 30024-30031.
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