The MEKK1 PHD ubiquitinates TAB1 to activate MAPKs in response to cytokines

Unlike the other MAP3Ks, MEKK1 (encoded by Map3k1) contains a PHD motif. To understand the role of this motif, we have created a knockin mutant of mouse Map3k1 (Map3k1(mPHD)) with an inactive PHD motif. Map3k1(mPHD) ES cells demonstrate that the MEKK1 PHD controls p38 and JNK activation during TGF-, EGF and microtubule disruption signalling, but does not affect MAPK responses to hyperosmotic stress. Protein microarray profiling identified the adaptor TAB1 as a PHD substrate, and TGF-- or EGF-stimulated Map3k1(mPHD) ES cells exhibit defective non-canonical ubiquitination of MEKK1 and TAB1. The MEKK1 PHD binds and mediates the transfer of Lys63-linked poly-Ub, using the conjugating enzyme UBE2N, onto TAB1 to regulate TAK1 and MAPK activation by TGF- and EGF. Both the MEKK1 PHD and TAB1 are critical for ES-cell differentiation and tumourigenesis. Map3k1(mPHD/+) mice exhibit aberrant cardiac tissue, B-cell development, testis and T-cell signalling.