PDF file: EURASNET Annual Report 2008

Participant 15 – Javier F. Cáceresa) Work performed during the period• WP8We are using the CLIP protocol (Cross-linking and immunoprecipitation) toidentify RNA targets of SR family proteins (SF2/ASF), hnRNP A1 and theExon junction component, Acinus. In all cases, use of CLIP has beencomplemented with subcellular fractionation to identify nuclear, cytoplasmic andpolysomal targets of these splicing factors. This has resulted in a publicationdescribing nuclear and cytoplasmic RNA targets of SF2/ASF (Sanford et al. (2008)).• WP10I am the lead participant in WP10. In collaboration with Angus Lamond’s labwe have used FRET/FLIM microscopy to study the interaction between SRproteins and splicing components that are bound at the 5’ or 3’ splice site.FRET imaging by means of fluorescence lifetime imaging microscopy (FLIM)allowed the mapping of these interactions to specific sites in the nucleus. Thishas resulted in a joint EURASNET-member publication (Ellis et al., 2008).We have also uncovered the mechanism by which SF2/ASF activates translation. Wehave shown that SF2/ASF promotes translation initiation by recruiting components ofthe mTOR signaling pathway that enhance phosphorylation of 4E-BP, a competitiveinhibitor of cap-dependent translation, resulting in its inactivation (Michlewski et al.2008)b) Major cost items with justificationParticipant 15 Type of expenditure Budget total costspersonnel 146.14Consumables & Other 19,940.44trainingequipmenttravel 7,635.29= total direct costs 27,721.87+ overhead 155.82= total eligible costs 27,877.69Major cost items• ResearchConsumables €19,940.45Travel: 7,635.29 (some are itemized below)Keystone MeetingJavier Caceres: 1253.02Berlin TripD Longman: 1,007.56RNA Meeting, BerlinJavier Caceres: 1116.98167