Mechanism and In Vitro Pharmacology of TAK1 Inhibition by (5Z)-7-Oxozeaenol

Abstract

Transforming growth factor-β activated kinase-1 (TAK1) is a member of the mitogen-activated protein kinase kinase kinase (MAP3K) family that regulates several signaling pathways including NF-κB signal transduction and p38 activation. TAK1 deregulation has been implicated in human diseases including cancer and inflammation. Here, we show that, in addition to its kinase activity, TAK1 has intrinsic ATPase activity, that (5Z)-7-Oxozeaenol irreversibly inhibits TAK1, and that sensitivity to (5Z)-7-Oxozeaenol inhibition in hematological cancer cell lines is NRAS mutation status and TAK1 pathway dependent. X-ray crystallographic and mass spectrometric studies showed that (5Z)-7-Oxozeaenol forms a covalent complex with TAK1. Detailed biochemical characterization revealed that (5Z)-7-Oxozeaenol inhibited both the kinase and the ATPase activity of TAK1 following a bi-phase kinetics, consistent with the irreversible inhibition mechanism. In DoHH2 cells, (5Z)-7-Oxozeaenol potently inhibited the p38 phosphorylation driven by TAK1, and the inhibition lasted over 6 h after withdrawal of (5Z)-7-Oxozeaenol. Profiling (5Z)-7-Oxozeaenol in a panel of hematological cancer cells showed that sensitive cell lines tended to carry NRAS mutations and that genes in TAK1 regulated pathways were enriched in sensitive cell lines. Taken together, we have elucidated the molecular mechanism of a TAK1 irreversible inhibitor and laid the foundation for designing next generation TAK1 irreversible inhibitors. The NRAS-TAK1-Wnt signaling network discerned in our study may prove to be useful in patient selection for TAK1 targeted agents in hematological cancers.

Innate immunity kinase TAK1 phosphorylates Rab1 on a hotspot for posttranslational modifications by host and pathogen

“…In HEK-293 cells stably expressing Flag-Rab1, overexpression of full-length V5-TAK1 and activating partner Myc-TAB1 (24) led to increased phosphorylation of Rab1 T75. Addition of a TAK1-selective inhibitor, 5z-7-oxozeaenol (25), eliminated phosphorylation of Rab1 (Fig. 2D).…”

TGFβ-activated Kinase 1 (TAK1) Inhibition by 5Z-7-Oxozeaenol Attenuates Early Brain Injury after Experimental Subarachnoid Hemorrhage

“…10), which displays Ͼ33-and Ͼ62-fold selectivity over MEKK1 and MEKK4, respectively (31). Mechanistically, OZ forms a covalent complex with TAK1 and inhibits both the kinase and ATPase activity of TAK1 following a biphase kinetics (32). Importantly, OZ is a relatively safe drug found in vivo and has been previously reported to cross the blood-brain barrier (17,33,34).…”

The Activation of IL-1-Induced Enhancers Depends on TAK1 Kinase Activity and NF-κB p65

“…This compound covalently binds to C174 within the ATP-binding pocket of TAK1, allowing fast inhibition of TAK1 enzymatic activity in cultured cells (Wu et al, 2013). 5Z-7-oxozeaenol blocked the vast majority of IL-1-inducible genes ( Figure S3A; Table S2).…”