Abstracts (poster) - Wissenschaft Online

Silke Götze, Sonja Sievers, Oliver Müller
Epigenetic regulation in the Wnt signalling pathway
Epigenetic mechanisms are involved in controlling gene expression. The two main
mechanisms of epigenetic gene regulation are DNA methylation and histone
modification, which affect each other. Histone modifications like acetylation, methylation
and phosphorylation are known to be highly dynamic and are likely to be modulated by
signalling pathways. In contrast very little is known about dynamic changes of DNA
methylation, specifically if they occur and how they might be regulated. Therefore we
want to investigate, which epigenetic changes occur due to Wnt signalling and what
impact these changes have on expression of Wnt target genes.
For this approach we use DMH (differential methylation hybridisation) on a CpG island
microarray to identify changes in the DNA methylation. In a further step we will also
perform a ChIP (chromatin immunoprecipitation) on chip analysis to detect changes in
the histone H3 acetylation.
In a first step we evaluated a proper Wnt model system with a Wnt inactive and a Wnt
active state. We used western blotting and RNA expression analysis to find a suitable
model system. DMH with this model system led to a number of candidate genes, which
showed differential DNA methylation on the microarray. We performed bisulfite
sequencing of some candidate genes but we could not verify the microarray results. The
high number of false positive results was likely caused by incomplete restriction during
DMH. Thus we established internal controls to monitor the restriction during repetition of
the DMH experiment.
contact:
M.Sc. Silke Götze
MPI für molekulare Physiologie
Abteilung Strukturbiologie
Silke.Goetze@mpi-dortmund.mpg.de
Otto-Hahn-Strasse 11
44227 Dortmund (Deutschland)