Impact of WIN site inhibitor on the WDR5 interactome

The chromatin-associated protein WDR5 is a promising pharmacological target in cancer, with most drug discovery efforts directed against an arginine-binding cavity in WDR5 called the WIN site. Despite a clear expectation that WIN site inhibitors will alter the repertoire of WDR5 interaction partners, their impact on the WDR5 interactome remains unknown. Here, we use quantitative proteomics to delineate how the WDR5 interactome is changed by WIN site inhibition. We show that the WIN site inhibitor alters the interaction of WDR5 with dozens of proteins, including those linked to phosphatidylinositol 3-kinase (PI3K) signaling. As proof of concept, we demonstrate that the master kinase PDPK1 is a bona fide high-affinity WIN site binding protein that engages WDR5 to modulate transcription of genes expressed in the G2 phase of the cell cycle. This dataset expands our understanding of WDR5 and serves as a resource for deciphering the action of WIN site inhibitors.

Automated summary

This study demonstrates how WIN site inhibitors alter the WDR5 interactome by using quantitative proteomics, showing that the inhibitors change the interaction of WDR5 with numerous proteins, including those related to the PI3K signaling pathway. The research also proves that the master kinase PDPK1 is a high-affinity binding protein for the WIN site, modulating gene transcription in the G2 phase of the cell cycle.