Browsing by Author "Pablo Huidobro Toro, J."

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    The 5'-untranslated region of the mouse mammary tumor virus mRNA exhibits cap-independent translation initiation
    (OXFORD UNIV PRESS, 2010) Vallejos, Maricarmen; Ramdohr, Pablo; Valiente Echeverria, Fernando; Tapia, Karla; Rodriguez, Felipe E.; Lowy, Fernando; Pablo Huidobro Toro, J.; Dangerfield, John A.; Lopez Lastra, Marcelo
    In this study, we demonstrate the identification of an internal ribosome entry site (IRES) within the 5'-untranslated region (5'-UTR) of the mouse mammary tumor virus (MMTV). The 5'-UTR of the full-length mRNA derived from the infectious, complete MMTV genome was cloned into a dual luciferase reporter construct containing an upstream Renilla luciferase gene (RLuc) and a downstream firefly luciferase gene (FLuc). In rabbit reticulocyte lysate, the MMTV 5'-UTR was capable of driving translation of the second cistron. In vitro translational activity from the MMTV 5'-UTR was resistant to the addition of m(7)GpppG cap-analog and cleavage of eIF4G by foot-and-mouth disease virus (FMDV) L-protease. IRES activity was also demonstrated in the Xenopus laevis oocyte by micro-injection of capped and polyadenylated bicistronic RNAs harboring the MMTV-5'-UTR. Finally, transfection assays showed that the MMTV-IRES exhibits cell type-dependent translational activity, suggesting a requirement for as yet unidentified cellular factors for its optimal function.