The tumour suppressor p53 (TP53), controls a wide range of genes that support cell cycle arrest, senescence, apoptosis, and DNA repair etc. Mutation of p53 is found in almost 50% of human cancers. Mutant p53 (mutp53) manifests itself through loss-of-function, dominant-negative and gain-of function (GOF) mechanisms, which prevent the cellular functions. One of the important GOF mechanisms of mut-p53 is the interaction with nuclear factor erythroid-derived 2-like-2 (Nrf2). Nrf2 is a crucial transcription factor for the expression of antioxidant genes during oxidative stress. Recent studies reported that mutp53/Nrf2 complex can upregulate proteasomal subunit genes (PSMA2 and PSMC1).
While the oncogenic role of R280K and R175H mutations in p53 were shown to mediate its prosurvival effects through p53/Nrf2 assembly, how these specific mutations (R280K and R175H) result in the interaction of mutp53 and Nrf2, is still not known. Therefore, in this study we plan to investigate the interaction of mutp53 and Nrf2 at the structural level, by using computational and experimental tools. As no comprehensive domain-wise data of the tertiary structure Nrf2 has been reported till date, we plan to use the computationally predicted Nrf2 protein structure for our study. In our experimental studies we plan to make specific mutant forms of p53 and different domain-wise constructs of NRF2 with GST-tag. Next, we will express and purify different domains of Nrf2, followed by Co-IP and Pull-Down assays to find out the interaction site between these two proteins. Finally, based on the interaction studies we plan to design a suitable cyclic peptide inhibitor to prevent mutp53/Nrf2 interaction.