RIBOSOME BIOGENESIS IN ZEBRAFISH DEVELOPMENT: NEW APPROACHES TO STUDY RIBOSOMOPATHIES

MOUGUELAR, VALERIA S.; PORCEL DE PERALTA, MAURO S.; CARNEVALE, MATIAS; CALCATERRA, NORA B; COUX, GABRIELA

Mar del Plata

Congreso; 51st Annual Meeting Argentine Society for Biochemistry and Molecular Biology; 2015

SAIB

Collectively, ribosomopathies are caused by defects in ribosome biogenesis. Although these disorders encompassdeficiencies in a ubiquitous and fundamental process, the clinical manifestations are highly variable and typicallydisplay tissue specificity. An example is the Treacher Collins Syndrome (TCS), a craniofacial development disorderdue to mutations in the TCOF1 gene, which encodes the protein TREACLE. Previously we have developed azebrafish TCS-like model through the injection of translation-blocking morpholinos against the TCOF1 ortholog(nolc1). Our aims here were: i) to develop anti-nolc1 antibodies to deepen our studies in zebrafish TCS model, ii) todesign an efficient and simple method to measure ribosome biosynthesis in TCS-like fish embryos. Zebrafish Nterminalnolc1 was cloned in the pRSET A plasmid, expressed and purified by Ni-agarose chromatography. Therabbit polyclonal antibodies obtained were highly specific and allowed us to corroborate a diminished translation ofthe homolog of TREACLE. We designed specific primers for the 5?ETS and ITS1 regions of the 47S pre-rRNA.rRNA transcription was estimated by RT-qPCR at different developmental stages in normal and TCS-like zebrafishembryos. Our results highlight the necessity to perform a scan along embryo development to better detect theribosomal biosynthetic performance under a pathological condition as TCS