O-Glc-NAc modifications of RNA-binding proteins

O-GlcNAcylation is a post-translational dynamic modification of proteins that is becoming more widely recognized as a modification that can protect proteins from pathological aggregation in neurodegenerative diseases. However, it is unknown how this modification influences the RNA binding properties of the protein and its function. In the proposed project I will study the O-Glc-NAcylation of RNA-binding proteins (RBPs) associated with amyotrophic lateral sclerosis (ALS), and its effects on the RNA binding properties and thereby on the regulation of RNA processing. I will find O-GlcNAcylation sites of TDP-43, Fused-in-sarcoma (FUS), Ewing sarcoma protein (EWS), and TATA-box binding protein associated factor 15 (TAF15) and I will assess the increased amounts of modification after stimulation of the cells with high glucose or with O-Glc-NAcase inhibitor. I will study if their RNA interaction profiles change with increasing amounts of O-Glc-NAc modifications in differentiated cortical neurons. Later, I plan to implement in the company, a mass spectrometry method adapted to analyse the ALS-liked RBPs proteins, which will help the glycoscience community. This project will expand our understanding of the molecular mechanism of transcription regulation through protein O-Glc-NAcylation and can provide the necessary knowledge to understand ALS-related proteinopathies.