IRAK3 modulates downstream innate immune signalling through its guanylate cyclase activity.

L A Freihat, J I Wheeler, Aloysius Tze Wong, I Turek, D T Manallack, H R Irving

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30 Scopus citations


Interleukin-1 receptor associated kinase 3 (IRAK3) is a cytoplasmic homeostatic mediator of inflammatory responses and is potentially useful as a prognostic marker in inflammation. IRAK3 inhibits signalling cascades downstream of myddosome complexes associated with toll like receptors. IRAK3 contains a death domain that interacts with other IRAK family members, a pseudokinase domain and a C-terminus domain involved with tumour necrosis factor receptor associated factor 6 (TRAF6). Previous bioinformatic studies revealed that IRAK3 contained a guanylate cyclase centre in its pseudokinase domain but its role in IRAK3 action is unresolved. We demonstrate that wildtype IRAK3 is capable of producing cGMP. Furthermore, we show that a specific point mutation in the guanylate cyclase centre reduced cGMP production. Cells containing toll like receptor 4 and a nuclear factor kappa-light-chain-enhancer of activated B cells (NFĸB) reporter system were transfected with IRAK3 or mutant IRAK3 proteins. Cell-permeable cGMP treatment of untransfected control cells suppresses downstream signalling through modulation of the NFĸB in the presence of lipopolysaccharides. Cells transfected with wildtype IRAK3 also suppress lipopolysaccharide induced NFĸB activity in the absence of exogenous cGMP. Lipopolysaccharide induced NFĸB activity was not suppressed in cells transfected with the IRAK3 mutant with reduced cGMP-generating capacity. Whereas in the presence of exogenously applied cell-permeable cGMP the IRAK3 mutant was able to retain its function by suppressing lipopolysaccharide induced NFĸB activity. Furthermore, increasing the amount of membrane permeable cGMP did not affect IRAK3's ability to reduce NFĸB activity. These results suggest that cGMP generated by IRAK3 may be involved in regulatory function of the protein where the presence of cGMP may selectively affect downstream signalling pathway(s) by modulating binding and/or activity of nearby proteins that interact in the inflammatory signalling cascade.
Original languageEnglish (US)
JournalScientific reports
Issue number1
StatePublished - Oct 31 2019

Bibliographical note

KAUST Repository Item: Exported on 2020-10-01
Acknowledgements: Support from the Australian Research Council’s Discovery funding scheme (project number DP110104164) and the Monash Institute of Pharmaceutical Science Large Grant Support Scheme is gratefully acknowledged. Research was sponsored by the Army Research Office and was accomplished under Grant Number W911NF-17-1-0303. The views and conclusions contained in this document are those of the authors and should not be interpreted as representing the official policies, either expressed or implied, of the Army Research Office or the U.S. Government. LAF received a scholarship from the Monash Institute of Pharmaceutical Sciences. AW is supported by the National Natural Science Foundation of China (Grant no. 31850410470) and the Zhejiang Provincial Natural Science Foundation of China (Grant no. LQ19C130001).


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