Phosphorylation-mediated regulation of the NF-kB subunit c-Rel

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Nuclear Factor kappa B (NF-kB) is a central regulator of the immune, stress and inflammatory responses, and additionally plays key roles in cell differentiation and survival. Aberrant regulation of NF-kB is also associated with many pathological conditions, including inflammatory diseases and cancer.Mass spectrometric and cell biological techniques will be used to identify and characterise the functions of post-translational modifications (PTMs) of the c-Rel NF-kB subunit in B cells. Biological mass spectrometry will be used in PTM site identification, protein quantification and top-down proteomics. Biochemical/cell biology based approaches will be used to elucidate the biological role/transcription mediation of the c-Rel modification sites that are found. This includes cell culture, nucleofection-based transfection of B-cell lymphoma cell lines, Western blotting, q-PCR analysis of c-Rel target gene expression and analysis of cell viability. As the project progresses and more information is acquired about c-Rel and its interactions, functional experiments using c-Rel mutant expression plasmids will be used in order to gain more insight into the roles of c-Rel.