PDF file: EURASNET Annual Report 2008

10 JUSTIFICATION OF MAJOR COST ITEMS AND RESOURCESParticipant 1A – Reinhard Lührmann, Coordinatora) Work performed during the period• WP2The Lührmann lab provided protocols, expertise and biological research material to severalnetwqork members.• WP9We affinity-purified individual, human spliceosomal complexes formed in HeLa splicingextract under identical physiological conditions (150 mM salt) and determined their proteincomposition by mass spectrometry.We developed a novel 2D electrophoretic procedure utilizing principles previously used forthe separation of ribosomal proteins, but with major modifications.In feasibility studies we treated tri-snRNP particles with Sulfo-MBS and separated theproteins by 2D gel electrophoresis.In another feasibility study we investigated the protein-protein contacts in the purified humanPrp19/CDC5L.We established HeLa S3 cell lines that stably express FLAG/HA-tagged AD002 or SPF27. 2Delectron microscopy of immunoaffintiy purified 14S Prp19/CDC5L complexes from both celllines revealed compact and well-defined particles of similar size. Protein-protein interactionswithin the Prp19/CDC5L complex were investigated by chemical cross-linking with Sulfo-MBS.• WP10We performed a large scale phosproteome analysis of the different spliceosomal complexesderived from human cells.• WP14We could show that several small-molecule inhibitors of protein acetylation/deacetylationblock the splicing cycle.• WP16Organized the January 2008 IFM in Berlin. Supervised all IFM, workshop and meetingsevents.• WP17The Lührmann lab provided extensive training for other EURASNET labs within the travelbursary program.• WP20We identified the mechanism of action of a small molecule splicing inhibitor.• WP22The Göttingen based management team supervised all research, integration and disseminationactivities of the network, organized the 2008 report, steering committee meetings, handled alltravel reimbursements and financial transactions. It provided support for all legal and financialproblems of the network and served as the hub for network information.b) Major cost items with justification128