SRA

The WIN site of WDR5 is a druggable pocket that is crucial for WDR5 protein function and carries therapeutic potential for treating cancer. This study evaluates the protein interactions affected by small molecule blockade of the WIN site of WDR5. We find that PDPK1 directly binds the WIN site of WDR5, and we investigate this newfound interaction through proteomic, biochemical, and genomic methods. Overall design: RNA-seq was used to monitor transcriptional changes resulting from targeting PDPK1 and WDR5. Degradation systems for PDPK1 and WDR5 were generated in U2OS cells. RNA-seq was performed in these cell lines after treatments for 24 hours with DMSO vehicle control or 500 nM dTAG47 to induce degradation. In HEK293 cells the PDPK1 R3A point mutation was introduced using CRISPR genome editing and retroviral transductions. RNA-seq was performed in these engineered cells.