PDF file: EURASNET Annual Report 2008

Participant 10A – Giuseppe Biamontia) Work performed during the period• WP2 Sharing Resources, Technology and Reliable Protocols – Ian Eperon• WP7 Molecular Characterization of Splicing Substrates – Christiane Branlant.10A identified elements in the SF2/ASF transcripts that control the unproductivesplicing of the pre-mRNA. These sequence elements are bound by the RNA binding protein Sam68 andare close to the ultra-conserved DNA elements.• WP8 Genome-wide Analyses of Splicing Regulation – Chris Smith10A used splicing–sensitive micro-arrays to investigate changes of splicing profileduring EMT. They showed that, in general, transcripts for splicing factors are major targets of splicingreprogramming. In particular this analysis proved that transcripts for all SR factors and for a subset ofhnRNP proteins undergo unproductive splicing events that introduce premature stop codons and directRNA to degradation through the NMD pathway.• WP10 Post-translational Modification and Dynamic Regulation – Javier Cáceres10A showed that the phosphorylation of Sam68 by the ERK kinase promotesunproductive splicing of SF2/ASF transcripts leading to a decreased level of SF2/ASF protein. Thisfinding identifies a mechanism through which signal transduction cascades activated by externalstimuli may control the level of splicing factors and hence the splicing profile of several human genes.• WP12 Mis-splicing and Disease – Francisco Baralle10A showed that the RNA binding protein Sam68 controls the unproductive splicingof SF2/ASF transcripts. The expression of Sam68 is altered in cancer specimens. In all cases in whichSam68 is up-regulated there is an increased unproductive splicing of SF2/ASF transcripts. Moreover,the activity of Sam68 is controlled by the ERK kinase. These findings identify a mechanism throughwhich stimuli in the tumor microenvironment may induce unproductive splicing of transcripts forrelevant splicing factors.• WP13 Co-transcriptional Mechanisms of Alternative Splicing – KarlaNeugebauer10A investigated the fate of non-coding RNAs deriving from the transcription oflong arrays of Satellite III repetitive DNA. These RNAs remain associated with sites of transcriptionand are bound by splicing factors. However, instead of undergoing splicing events they are eventuallyprocessed into small RNAs.b) Major cost items with justificationParticipant 10A Type of expenditure budget total costspersonnel 27.268,92consumables 16.205,06trainingequipment 722,80travel 6041,38= total direct costs 50238,16+ overhead 21678,79adjustment* -3.472,69= total eligible costs 68.444,26*Adjustment due to overestimation of personnel costs in the previous years150