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The E3 ligase Itch negatively regulates inflammatory signaling pathways by controlling the function of the ubiquitin-editing enzyme A20

Nature Immunology volume 9, pages 254–262 (2008)Cite this article

Abstract

The ubiquitin-editing enzyme A20 is a critical negative regulator of inflammation and cytokine-mediated activation of the transcription factor NF-κB; however, little is known about the mechanisms of A20-mediated inactivation of signaling intermediates such as RIP1. Here we demonstrate that the regulatory molecule TAX1BP1 recruited the E3 ligase Itch to A20 via two 'PPXY' motifs. Itch was essential for the termination of tumor necrosis factor receptor signaling by controlling A20-mediated recruitment and inactivation of RIP1. Furthermore, the Tax oncoprotein of human T cell leukemia virus type I targeted this complex for inactivation by disrupting the interaction among TAX1BP1, A20 and Itch. Thus, our studies show a previously unappreciated complexity of A20 substrate recognition and inactivation whereby TAX1BP1 and Itch function as essential subunits of an A20 ubiquitin-editing complex.

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Figure 1: TAX1BP1 requires two zinc-finger domains and associated PPXY motifs for inactivation of NF-κB and Jnk.
Figure 2: TAX1BP1 interacts with RIP1 and Itch through zinc-finger domains and PPXY motifs.
Figure 3: Itch is required for the termination of TNF- and IL-1-mediated NF-κB and Jnk signaling.
Figure 4: A20 requires Itch for recruitment to RIP1, deubiquitination of RIP1 and inhibition of NF-κB.
Figure 5: Itch promotes RIP1 degradation in an A20- and TAX1BP1-dependent way.
Figure 6: HTLV-I Tax disrupts the interaction among TAX1BP1, Itch and A20 and inactivates the targeting and function of A20.
Figure 7: The phenotype of Itch−/− mice is lymphocyte autonomous.

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Acknowledgements

We thank S. Sun, G. Barber, K. Tolba and C. Vincenz for reagents; A. Angers (University of Montreal) for plasmid encoding Flag-Itch; J. Tschopp (University of Lausanne) for Flag–c-FLIP(L); P. Storz (Mayo Clinic, Jacksonville) for pSUPER and pSUPER A20-specific siRNA; W. Greene (Gladstone Institute for Virology and Immunology) for Jurkat Tax Tet-on cells; and D.A. Swing, R. Koogle and J.B. Lea for assistance with mouse husbandry. Supported by the University of Miami Scientific Awards Committee.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Sylvester Comprehensive Cancer Center, The University of Miami, Miller School of Medicine, Miami, 33136, Florida, USA

    Noula Shembade, Nicole S Harhaj, Kislay Parvatiyar & Edward W Harhaj

  2. Mouse Cancer Genetics Program, Center for Cancer Research, National Cancer Institute, Frederick, 21702, Maryland, USA

    Neal G Copeland, Nancy A Jenkins & Lydia E Matesic

  3. Department of Biological Sciences, University of South Carolina, Columbia, 29208, South Carolina, USA

    Lydia E Matesic

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  1. Noula Shembade
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Contributions

N.S., N.S.H., K.P. and L.E.M. did experiments and analyzed data; N.G.C., N.A.J. and L.E.M. provided essential reagents; L.E.M. assisted in writing the manuscript; and E.W.H. designed and supervised the experiments, analyzed data and wrote the manuscript.

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Correspondence to Edward W Harhaj.

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Shembade, N., Harhaj, N., Parvatiyar, K. et al. The E3 ligase Itch negatively regulates inflammatory signaling pathways by controlling the function of the ubiquitin-editing enzyme A20. Nat Immunol 9, 254–262 (2008). https://doi.org/10.1038/ni1563

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