PERSONAL DE APOYO
ROJAS hernan Javier
congresos y reuniones científicas
Título:
Post transcriptional silencing of RNAses during phosphate starvation in Nicotiana
Autor/es:
ROJAS HERNAN JAVIER; GOLDRAIJ ARIEL; ESCOBAR ELIANA
Lugar:
Mar del Plata
Reunión:
Congreso; LI Reunión Anual de la Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular (SAIB); 2015
Institución organizadora:
Reunión Anual de la Sociedad Argentina de Investigaciones en Bioquímica y Biología Molecular (SAIB)
Resumen:
Ribonucleases (RNases) T2 are endonucleolytic enzymes that catalyze the cleavage of single-strand RNA producing mononucleotides with a terminal 3' phosphate (Pi). In plants, different classes of RNases T2 are induced in a variety of stress scenarios, including Pi starvation. The induction of these RNases contributes to Pi mobilization from nucleic acid sources. In Nicotiana alata, class III NnSR1 (Nicotiana non S-RNase1) and class I S-like-RNase NE are, thus far, the only RNases induced for Pi mobilization. NnSR1 is localized in endoplasmic reticulum of root cells while RNase NE is secreted to the rizosphere to scavenge extracellular RNA sources. To discern whether the induction of thesetwo RNases is simultaneous or sequential, as well as the Pi threshold triggering such induction, plants were cultivated under Pi limiting concentrations. Early induction of NnSR1, assayed by western blot and in gel RNase activity,suggested that the intracellular RNA may be the first source of Pi used by the cell under Pi stress. To understand the functional role of these RNases, a post transcriptional silencing strategy was developed. An hypervariable region ofboth RNase NE and NnSR1was selected to clone into a binary vector. Preliminar PCR, restriction analysis and subsequent plasmid DNA sequencing indicated successful cloning to express the hairpin RNA required for silencingRNase expression.