Stitched peptides as potential cell permeable inhibitors of oncogenic DAXX protein

Jelinska, C.; Kannan, S.; Winnerdy, F. R.; Frosi, Y.; Ramlan, S. R.; Lakshminarayanan, R.; Brown, C. J.; Phan, A.-T.; Rhodes, D.; Verma, C. S.

doi:10.1101/2022.09.25.508451

bioRxiv · 10.1101/2022.09.25.508451

Stitched peptides as potential cell permeable inhibitors of oncogenic DAXX protein

Jelinska, C.·Kannan, S.·Winnerdy, F. R.·Frosi, Y.·Ramlan, S. R.·Lakshminarayanan, R.·Brown, C. J.·Phan, A.-T.·Rhodes, D.·Verma, C. S.

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Abstract

The death domain associated protein 6 (DAXX) is frequently upregulated in a number of common cancers where its suppression has been linked to reduced tumour progression. As a master regulator protein, with >70 reported protein interaction partners, the role of DAXX in its oncogenecity remains unclear. We designed and developed a set of novel stapled/stitched peptides that target a surface on the N-terminal helical bundle domain of DAXX which is the anchor-point for binding to multiple interaction partners (including Rassf1C, P53, Mdm2 and ATRX) and also for the auto regulation of the DAXX N-terminal SUMO interaction motif (SIM). We demonstrate that these peptides bind to and inhibit DAXX with an affinity higher than those reported for the known interaction partners and release the auto-inhibited SIM for interaction with SUMO-1. NanoBret assays show that the peptides enter cells and that their intracellular concentrations remain at nanomolar levels even after 24 hours, without causing membrane perturbation. Together our data suggest that these peptides are both tools for probing the molecular interactions of DAXX and potential precursors to the development of therapeutics.

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Jelinska, C., Kannan, S., Winnerdy, F. R., Frosi, Y., Ramlan, S. R., Lakshminarayanan, R., Brown, C. J., Phan, A.-T., Rhodes, D., Verma, C. S.. 2022-09-25. Stitched peptides as potential cell permeable inhibitors of oncogenic DAXX protein. https://doi.org/10.1101/2022.09.25.508451

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