SMYD3 is a histone methylase that di- and tri-methylates histone H3 at lysine 4. In normal conditions SMYD3 is highly expressed only in skeletal muscle and testis. We hypothesize that SMYD3 plays an important role in transcriptional regulation throughout myogenesis. We will use C2C12 myoblast and primary satellite cells as model systems to assess the contribution of SMYD3 to the transcriptional regulation of muscle specific genes. We will over-express and knock down SMYD3 to determine the impact of SMYD3 on the expression of myogenic markers and the ability of C2C12 cells to fuse into multinucleated myotubes. We will also determine functional mechanisms that account for a potential involvement of SMYD3 in myogenesis by employing chromatin immunoprecipitation experiments. Moreover, we will identify SMYD3 target genes in muscle cells by RNA microarray analysis and by the combination of the chromatin immunoprecipitation technique with massively sequencing technology (ChIp-seq). A further objective of the project will be to gain insights into the mechanisms underlying SMYD3 activity and its modulation, by exploiting in vitro and in vivo approaches.