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Nucleic Acids Research, 2003, Vol. 31, No. 10 2508-2513
© 2003 Oxford University Press

Translational control of Scamper expression via a cell-specific internal ribosome entry site

Davide De Pietri Tonelli1,2, Marija Mihailovich1,2, Raphaela Schnurbus1, Graziano Pesole3, Fabio Grohovaz1,2 and Daniele Zacchetti1

1 Cellular Neurophysiology Unit, Department of Neuroscience, San Raffaele Scientific Institute and 2 Vita-Salute San Raffaele University, Via Olgettina 58, I-20132 Milano, Italy and 3 Department of General Physiology and Biochemistry, University of Milano, I-20133 Milano, Italy

*To whom correspondence should be addressed. Tel: +39 02 2643 4812; Fax: +39 02 2643 4813; Email: zacchetti.daniele{at}hsr.it

The mRNA of Scamper, a putative intracellular calcium channel activated by sphingosylphosphocholine, contains a long 5' transcript leader with several upstream AUGs. In this work we have investigated the role this sequence plays in the translational control of Scamper expression. The cytosolic transcription machinery of a T7 RNA polymerase recombinant vaccinia virus was used to avoid artifacts arising from cryptic promoters or mRNA processing. Based on transient transfection experiments of dicistronic and bi-monocistronic plasmids expressing reporter genes, we present evidence that the 5' transcript leader of Scamper contains a functional internal ribosome entry site (IRES). Our data indicate that Scamper translation in Madin-Darby canine kidney cells is driven by a cap-independent mechanism supported by the IRES activity of its mRNA. Finally, the Scamper IRES appears to be the first IRES with specificity for kidney epithelial cells.


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