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J. Biol. Chem., Vol. 278, Issue 3, 1511-1517, January 17, 2003
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From the International Centre for Genetic Engineering and
Biotechnology, Padriciano 99, Trieste 34012, Italy and the
Using hybrid minigene experiments, we have
investigated the role of the promoter architecture on the regulation of
two alternative spliced exons, cystic fibrosis transmembrane regulator
(CFTR) exon 9 and fibronectin extra domain-A (EDB). A specific
alternative splicing pattern corresponded to each analyzed promoter.
Promoter-dependent sensitivity to cotransfected regulatory
splicing factor SF2/ASF was observed only for the CFTR exon 9, whereas
that of the EDB was refractory to promoter-mediated regulation.
Deletion in the CFTR minigene of the downstream intronic
splicing silencer element binding SF2/ASF abolished the specific
promoter-mediated response to this splicing factor. A systematic
analysis of the regulatory cis-acting elements showed that
in the presence of suboptimal splice sites or by deletion of exonic
enhancer elements the promoter-dependent sensitivity to
splicing factor-mediated inhibition was lost. However, the basal
regulatory effect of each promoter was preserved. The complex
relationships between the promoter-dependent sensitivity to
SF2 modulated by the exon 9 definition suggest a kinetic model of
promoter-dependent alternative splicing regulation that
possibly involves differential RNA polymerase II elongation.
Promoter Architecture Modulates CFTR Exon 9 Skipping*
§, and Laboratorio de Fisiología y Biología
Molecular, Departamento de Fisiología, Biología
Molecular y Celular, Facultad de Ciencias Exactas y Naturales,
Universidad de Buenos Aires, Ciudad Universitaria, Pabellón
II (C1428EHA) Buenos Aires, Argentina
*
This work was supported by Telethon-Italy Grant GGP02453 and
by a grant from the Associasione Italiana Ricerca Cancro (AIRC).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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