ARGONAUTE proteins transport routes and subcelular activities

ended

Small RNAs (sRNA) have emerged as major players in a multitude of pathways, controlling development and preserving genome integrity in plant and animals. To fulfill their biological function, sRNA are loaded into ARGONAUTE (AGO) proteins, together this AGO/sRNA complex recognizes and regulates the target genes. Depending on the organism and the sRNA pathway involved, AGOs have nuclear and/or the cytoplasmic localization and functions. Although globally AGO mediated processes are reasonably understood, the transport routes involved, their shuttling regulation and transient subcellular roles remain completely unknown. This proposal outlines a series of mutually reinforcing experiments to mechanistically study the transport routes and subcellular roles of AGO-sRNA complexes. In details, combining cell biology, RNA biochemistry, protein modeling, bioinformatics, deep-sequencing analyses, nuclear fractionation methods and single-molecule studies the proposed project aims to: 1) Identify key subcellular localization signals of AGO proteins; 2) Decipher AGO shuttling mechanism and their regulation; and 3) Dissect cytoplasmic and nuclear roles of AGO proteins on RNA pathways. The anticipated results will provide a crucial breakthrough in the eukaryotic RNA pathways and gene regulation fields. The interest in non-coding RNA action goes beyond mere mechanistic knowledge. Understanding non-coding RNA action is of paramount importance given the extraordinary potential held by the therapeutic, veterinary and agronomic exploitation of these molecules. As a result, the knowledge on the transport routes and subcellular roles of AGO-sRNA complexes obtained during the proposed project will provide an important contribution to European excellence and competitiveness in the field.