Differential regulation of NF-κB activation and function through RelA Ser42 and 45 phosphorylation (360G-Wellcome-206993_Z_17_Z)

NF-kappaB transcription factor activity can be regulated by a wide array of post-translational modifications that allow integration with parallel signalling pathways, leading to differential functionality in different contexts. Published data supports an important role for RelA Ser42 and Ser45 phosphorylation as regulators of NF-kappaB function but experiments performed to date have been limited and many questions remain unanswered. However, our analysis of the structure of a RelA/p50 heterodimer to DNA indicates that these residues are not close to the core NF-kappaB consensus binding site but rather are in proximity to the DNA flanking these sites. This suggests a potential modulatory effect on NF-kappaB DNA-binding rather than total inhibition, possibly regulating target gene specificity in a temporal manner following activation. This vacation studentship project will test this hypothesis and (1) Investigate whether RelA Ser42 and Ser45 mutations have gene specific effects on NF-kappaB regulated transcription; (2) Determine the effect of RelA Ser42 and Ser45 mutations on cell viability following NF-kappaB induction; (3) Develop a system using CRISPR/Cas9 mediated genome engineering to target the Ser42 and Ser45 residues in endogenous RelA. Data from these experiments will provide insight into regulation of NF-kappaB and its role in cancer and inflammatory diseases.