Fibroblast Growth Factor 2 Internal Ribosome Entry Site (IRES) Activity Ex Vivo and in Transgenic Mice Reveals a Stringent Tissue-specific Regulation

doi:10.1083/jcb.150.1.275

Published online 11 July 2000. doi:10.1083/jcb.150.1.275

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© The Rockefeller University Press, 0021-9525/2000/7/275/ $5.00
The Journal of Cell Biology, Volume 150, Number 1, July 10, 2000 275-281

Reports

Fibroblast Growth Factor 2 Internal Ribosome Entry Site (IRES) Activity Ex Vivo and in Transgenic Mice Reveals a Stringent Tissue-specific Regulation

Laurent Créancier^a, Dominique Morello^b, Pascale Mercier^c, and Anne-Catherine Prats^a
^a Institut National de la Santé et de la Recherche Médicale U397, Endocrinologie et Communication Cellulaire, Institut Fédératif de Recherche Louis Bugnard, C.H.U. Rangueil, 31403 Toulouse Cedex 04, France
^b Centre de Biologie du Développement, UMR 5547, Université Paul Sabatier, Bâtiment 4R3, 118 Route de Narbonne, 31062 Toulouse Cedex 04, France
^c Institut de Pharmacologie et Biologie Structurale, UPR 9062, 31077 Toulouse Cedex 04, France

Correspondence to: Anne-Catherine Prats, Institut National de la Santé et de la Recherche Médicale U397, Endocrinologie et Communication Cellulaire, Institut Fédératif de Recherche Louis Bugnard, C.H.U. Rangueil, 31403 Toulouse Cedex 04, France. Tel:33-5-61-32-21-42 Fax:33-5-61-32-21-41 E-mail:pratsac{at}rangueil.inserm.fr.

Fibroblast growth factor 2 (FGF-2) is a powerful mitogen involvedin proliferation, differentiation, and survival of various cellsincluding neurons. FGF-2 expression is translationally regulated;in particular, the FGF-2 mRNA contains an internal ribosomeentry site (IRES) allowing cap-independent translation. Here,we have analyzed FGF-2 IRES tissue specificity ex vivo and invivo by using a dual luciferase bicistronic vector. This IRESwas active in most transiently transfected human and nonhumancell types, with a higher activity in p53 -/- osteosarcoma andneuroblastoma cell lines. Transgenic mice were generated usingbicistronic transgenes with FGF-2 IRES or encephalomyocarditisvirus (EMCV) IRES. Measurements of luciferase activity revealedhigh FGF-2 IRES activity in 11-d-old embryos (E11) but not inthe placenta; activity was high in the heart and brain of E16.FGF-2 IRES activity was low in most organs of the adult, butexceptionally high in the brain. Such spatiotemporal variationswere not observed with the EMCV IRES. These data, demonstratingthe strong tissue specificity of a mammalian IRES in vivo, suggesta pivotal role of translational IRES- dependent activation ofFGF-2 expression during embryogenesis and in adult brain. FGF-2IRES could constitute, thus, a powerful tool for gene transferin the central nervous system.

Key Words: central nervous system, development, FGF, in vivo, translationalcontrol

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