It is known that glycogen synthase kinase 3beta (GSK3beta) plays a role in splice site selection (Hernández et al., 2004). Additionally it could be shown that protein phosphatase 1 (PP1), which dephosphorylates and thus activates GSK3beta, influences alternative splicing (Novoyatleva et al., 2008). Consequently we are interested on direct effects of GSK3beta on alternative splicing.

   I will elaborate the very likely binding of GSK3beta to the splicing factor transformer2-beta1 (tra2beta1). I will also determine the function of a GSK3beta-tra2beta1-complex and the consequences of GSK3beta activation on the phosphorylation status of tra2beta1 and the ability of tra2beta1 to bind to RNA.

I am using different techniques, like analytical ultracentrifugation and electromobility shift assay as well as classical immunoprecipitation and cell stainings to demonstrate binding and colocalisation.

Functionally we will determine the effects of GSK3beta on splice site selection in minigenes, which were shown to be regulated by tra2beta1 and PP1 (Novoyatleva et al., 2008). Using the in vivo splicing assay to clarify these effects, we will gain a deeper insight into the regulation of splice site seletion by the well-known signal transduction proteins PP1 and GSK3beta.

Mail Helen Hager