Unannotated microprotein EMBOW regulates the interactome and chromatin and mitotic functions of WDR5

The conserved WD40-repeat protein WDR5 interacts with multiple proteins both inside and outside the nucleus. However, it is currently unclear whether and how the distribution of WDR5 between complexes is regulated. Here, we show that an unannotated microprotein EMBOW (endogenous microprotein binder of WDR5) dually encoded in the human SCRIB gene interacts with WDR5 and regulates its binding to multiple interaction partners, including KMT2A and KIF2A. EMBOW is cell cycle regulated, with two expression maxima at late G1 phase and G2/M phase. Loss of EMBOW decreases WDR5 interaction with KIF2A, aberrantly shortens mitotic spindle length, prolongs G2/M phase, and delays cell proliferation. In contrast, loss of EMBOW increases WDR5 interaction with KMT2A, leading to WDR5 binding to off-target genes, erroneously increasing H3K4me3 levels, and activating transcription of these genes. Together, these results implicate EMBOW as a regulator of WDR5 that regulates its interactions and prevents its off-target binding in multiple contexts. [Display omitted] •The transcript leader of SCRIB encodes a WDR5-binding microprotein, EMBOW•EMBOW is expressed at specific points in the cell cycle•EMBOW binding alters the WDR5 interactome•EMBOW loss dysregulates WDR5 binding to off-target genes and the mitotic spindle Chen et al. show that human SCRIB dually encodes an unannotated, cell-cycle-regulated microprotein, EMBOW. EMBOW interacts with WDR5 and regulates its binding to multiple interaction partners, including KMT2A and KIF2A, safeguarding WDR5 on-target binding on chromatin and the mitotic spindle during the cell cycle.