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

Davide de Pietri Tonelli, Marija Mihailovich, Raphaela Schnurbus, Graziano Pesole, Fabio Grohovaz, Daniele Zacchetti

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)2508-2513
Number of pages6
JournalNucleic Acids Research
Volume31
Issue number10
DOIs
Publication statusPublished - May 15 2003

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Messenger RNA
Madin Darby Canine Kidney Cells
Vaccinia virus
Calcium Channels
Reporter Genes
Artifacts
Transfection
Plasmids
Epithelial Cells
Kidney
Internal Ribosome Entry Sites
bacteriophage T7 RNA polymerase
sphingosine phosphorylcholine

ASJC Scopus subject areas

  • Genetics

Cite this

Translational control of Scamper expression via a cell-specific internal ribosome entry site. / de Pietri Tonelli, Davide; Mihailovich, Marija; Schnurbus, Raphaela; Pesole, Graziano; Grohovaz, Fabio; Zacchetti, Daniele.

In: Nucleic Acids Research, Vol. 31, No. 10, 15.05.2003, p. 2508-2513.

Research output: Contribution to journalArticle

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