Translational control of reaper: a proapoptotic gene of Drosophila

PAULUCCI EZEQUIEL; FERRERO PAOLA; RIVERA POMAR ROLANDO

Puerto Madryn

Congreso; 46 Annual Meeting Argentine Society for Biochemistry and Molecular Biology; 2010

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

 Translational control is a key step in gene expression regulation during apoptosis. Proapoptotic genes of Drosophila, reaper(rpr), hid, grim and sickle, are downregulated by the miRNA family miR-  miRNAs can act on different mRNAs and the same mRNA can be target of different miRNAs. In this complex scenario the differences in target recognition could result in different regulators recruited for different mRNA fates. We aim to the identification of regulatory factors controlling the cap-independent translation initiation of rpr mRNA. In this study we present the action of miRNAs on rpr mRNA translation. We designed a minirpr- Myc tagged gene for biochemical and functional studies in the natural context of the mRNA 5? and 3?UTRs, avoiding the use of reporter genes with tandem copies of miRNA targets. For translational regulation assay, S2 Drosophila cells were transfected with the synthetic mRNAs in two different conditions, with or without miR2 pre-hybridized. ELISA results have shown a down regulation effect for the in vivo translation of minigen transcript linked to miR2. Proteomic analysis has allowed identify several factors assembled in the 5? UTR of reaper. iRNA assay was designed to determinate feasible roles in the regulation machinery. Recent proteomic analysis with mini-transcripts provides interesting data about miRNA modulators and regulation of translation.