PDF file: EURASNET Annual Report 2008

Participant 29 – Frédéric Allaina) Work performed during the period•WP7We recently determined the structure of the RRM of tra2β in complex with AAGAAU using NMRspectroscopy. The structure showed that all six nucleotides are in contact with the RRM but also with theN and C-terminal extension. Key protein-RNA contacts have been tested by affinity measurement usingITC and by splicing assays (in collaboration with the Stamm lab) on mutant proteins.We also determined the 3D solution structure of the 3 RRMs of hnRNP F in complex with AGGGAU.The structure reveals a new mode of RNA binding for all three RRMs, since for each of them, the β-sheet surface is not used for RNA binding as is usually the case. These data explain how a G-tract ofthree consecutive Gs is recognized by hnRNP F. The structure also suggests mechanisms of action forhnRNP F. We showed that hnRNP F can unfold RNA structures (RNA stem-loop or G-quadruplex). Inbinding G-tract, hnRNP F could free key cis-acting element in contact with the G-tract thereforeactivating or repressing splicing.b) Major cost items with justificationParticipant 1A Type of expenditure Budget total costspersonnel 0consumables 5.716,35training 0equipment 0travel 422.30= total direct costs 6.138,65+ overhead 1.227.72= total eligible costs 7.366,37Major cost items• ResearchConsumablesOligonucleotides and laboratory supplies5.716,35 EuroTravel• Berlin422.30 EuroAll funds were spent on research, namely on consumables and travels. Consumables includedoligonucleotide synthesis, chemicals, restriction and modification enzymes. Travels included trips toBerlin in the context of meeting between Eurasnet members.189