O-GlcNAcylation of the transcription machinery in prostate cancer cells

O-GlcNAcylation is a post-translational protein modification overexpressed in the most aggressive prostate cancer, which is the most common cancer type amongst men in Finland. O-GlcNAcylation has been found to play a role in gene transcription as an alternative protein modification to phosphorylation by transcription-associated cyclin-dependent kinases (TCDK). The aim of this thesis is to study the O-GlcNAcylation of transcriptional machinery in prostate cancer cells when inhibiting each TCDK activity. Cancer cells were first treated with on-target doses of TCDK inhibitors, followed by enrichment of O-GlcNAcyted proteins using lectin pulldown and investigating transcriptional machinery proteins levels by performing western blot. Results revealed a significant increase of CDK9 O-GlcNAcylation after inhibiting CDK9 activity in all used prostate cell lines. These findings highlight the important and yet not well-known role of O-GlcNAcylation in gene transcription in prostate cancer cells.