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J. Biol. Chem., Vol. 277, Issue 39, 35808-35814, September 27, 2002
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From the U119 INSERM, Institute of Cancerology and Immunology of
Marseille, 27 Boulevard Lei Roure, F-13009, Marseille, France
Production of the two mRNAs encoding distinct
forms of 2'-5'-oligoadenylate synthetase depends on processing that
involves the recognition of alternative poly(A) sites and an internal
5'-splice site located within the first 3'-terminal exon. The
resulting 1.6- and 1.8-kb mRNAs are expressed in fibroblast cell
lines, whereas lymphoblastoid B cells, such as Daudi, produce only the 1.8-kb mRNA. In the present study, we have shown that the 3'-end processing at the last 3'-terminal exon occurs independently of the
core poly(A) site sequence or the presence of regulatory elements. In
contrast, in Daudi cells, the recognition of the poly(A) site at the
first 3'-terminal exon is impaired because of an unfavorable sequence
context. The 3'-end processing at this particular location requires a
strong stabilization of the cleavage/polyadenylation factors, which can
be achieved by the insertion of a 25-nucleotide long U-rich motif
identified upstream of the last poly(A) site. Consequently, we
speculate that in cells expressing the 1.6-kb mRNA, such as
fibroblasts, direct or indirect participation of a specific mechanism
or cell type-specific factors are required for an efficient
polyadenylation at the first 3'-terminal exon.
The Cleavage/Polyadenylation Activity Triggered by a U-rich Motif
Sequence Is Differently Required Depending on the Poly(A) Site Location
at Either the First or Last 3'-Terminal Exon of the 2'-5' Oligo(A)
Synthetase Gene*
§,¶,
, and
*
This work was supported by grants from the Institut National
de la Santé et de la Recherche Médicale, from the
Association pour la Recherche sur le Cancer, and the Ligue Nationale
contre le Cancer.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.
Both authors contributed equally to this work.
§
Present address: Centre de Génétique
Moléculaire-CNRS, F-91000, Gif-Sur-Yvette, France.
¶
Present address: Cellectis S.A. 28 Rue du Dr. Roux, F-75 724, Paris Cedex 15, France.
Present address: Centre de Therapie Cellulaire et Genique,
Institut Paoli-Calmettes, F-13009, Marseille, France.
**
Present address: Innovation Moléculaire pour la Valorisation
et le Transfert-Institut de Biologie du Developpement de Marseille, Case 907 Campus de Luminy, F-13288, Marseille, France. To whom correspondence should be addressed. Tel.: 33-4-91-26-96-24;
Fax: 33-4-91-26-97-26; E-mail: benech@ibdm.univ-mrs.fr.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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