TIPTGSVSR

Biochemical characterization of RNA silencing mechanisms and their alteration by viral proteins in plant cell-free systems

Coordinatore	EIDGENOESSISCHE TECHNISCHE HOCHSCHULE ZURICH    more  Organization address address: Raemistrasse 101 city: ZUERICH postcode: 8092 contact info Titolo: Prof. Nome: Olivier Cognome: Voinnet Email: send email Telefono: 41446339360
Nazionalità Coordinatore	Switzerland [CH]
Totale costo	184˙709 €
EC contributo	184˙709 €
Programma	FP7-PEOPLE Specific programme "People" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013)
Code Call	FP7-PEOPLE-2012-IIF
Funding Scheme	MC-IIF
Anno di inizio	2013
Periodo (anno-mese-giorno)	2013-03-01   -   2015-02-28

 Partecipanti

#	participant	country	role	EC contrib. [€]
1	EIDGENOESSISCHE TECHNISCHE HOCHSCHULE ZURICH  Organization address address: Raemistrasse 101 city: ZUERICH postcode: 8092 contact info Titolo: Prof. Nome: Olivier Cognome: Voinnet Email: send email Telefono: 41446339360	CH (ZUERICH)	coordinator	184˙709.40

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 Obiettivo del progetto (Objective)

'Posttranscriptional gene silencing (PTGS) is a form of small (s)RNA-mediated gene regulation with important roles in development, stress responses, and antiviral defense in plants. PTGS effectors, called RNA-induced silencing complexes (RISCs), have ARGONAUTE (AGO) proteins bound to single-stranded (ss) guide sRNAs as core components. RISCs recognize target RNA displaying complementarity to guide sRNAs, and inactivate them by endonucleolytic cleavage (or ‘slicing’) or by slicer-independent translational repression. A precise dissection of the underlying mechanisms has been, however, hampered by a lack of plant cell-free systems allowing reconstruction of key PTGS steps in vitro. Recently, a cell-free AGO1-RISC assembly system was developed in Japan, using extracts from evacuolated tobacco BY-2 protoplasts (BYL) and allowed to establish that AGO1-RISC assembly relies on molecular chaperoning by HSP90 and co-factor CYP40. The proposed project will further deepen the understanding of plant RISC assembly mechanisms and decipher the molecular underpinnings of other PTGS steps including Dicer processing and RISC-mediated translational repression. In parallel, the BYL system will be used to investigate, in a strict comparative manner, the direct targets and modes of action of several plant viral suppressors of RNA silencing (VSRs). These proteins have been studied so far mostly in transgenic plants, where feedback PTGS regulations, intrinsically variable VSR expression levels and indirect effects have led to an inaccurate- and sometimes contradictory- view of their action. BYL will bypass these caveats and improve our knowledge of VSRs on antiviral silencing, but also of PTGS, as VSRs will be used to stabilize transient, and thus far inaccessible forms of protein-protein and protein-RNA complexes. Knowledge on plant cell-free systems implemented via the proposed project will finally contribute to European excellence and competitiveness in the field.'