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The p110δ isoform of the kinase PI(3)K controls the subcellular compartmentalization of TLR4 signaling and protects from endotoxic shock

Nature Immunology volume 13, pages 1045–1054 (2012)Cite this article

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A Corrigendum to this article was published on 19 July 2013

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Abstract

Lipopolysaccharide activates plasma-membrane signaling and endosomal signaling by Toll-like receptor 4 (TLR4) through the TIRAP-MyD88 and TRAM-TRIF adaptor complexes, respectively, but it is unclear how the signaling switch between these cell compartments is coordinated. In dendritic cells, we found that the p110δ isoform of phosphatidylinositol-3-OH kinase (PI(3)K) induced internalization of TLR4 and dissociation of TIRAP from the plasma membrane, followed by calpain-mediated degradation of TIRAP. Accordingly, inactivation of p110δ prolonged TIRAP-mediated signaling from the plasma membrane, which augmented proinflammatory cytokine production while decreasing TRAM-dependent endosomal signaling that generated anti-inflammatory cytokines (interleukin 10 and interferon-β). In line with that altered signaling output, p110δ-deficient mice showed enhanced endotoxin-induced death. Thus, by controlling the 'topology' of TLR4 signaling complexes, p110δ balances overall homeostasis in the TLR4 pathway.

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Figure 1: The p110δ isoform of PI(3)K is recruited to the LPS-activated TLR4 complex and controls TLR4 internalization in DCs.
Figure 2: The activity of p110δ in BMDCs is involved in the dissociation of TIRAP from the plasma membrane after stimulation with LPS.
Figure 3: PTEN regulates LPS-induced TIRAP degradation and TLR4 internalization in BMDCs, but SHIP does not.
Figure 4: 'Licensing' of calpain-induced TIRAP proteolysis by p110δ in BMDCs and J774 macrophages.
Figure 5: Inhibition of p110δ in BMDCs augments the production of proinflammatory cytokines while inhibiting IFN-β and the products of interferon-stimulated genes.
Figure 6: The kinase activity of p110δ in BMDCs is required for optimal IFN-β production through IRF3 and late activation of NF-κB and p38.
Figure 7: Inhibition of p110δ in mice augments endotoxin-mediated death.

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  • 08 April 2013

    In the version of this article initially published, the eighth author was identified incorrectly. The correct name is Inma M Berenjeno, and the initials should be I.M.B. in the Author Contributions section. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank R. Medzhitov (Yale University) for expression vectors encoding GST-TIRAP, GFP-TIRAP and GST–TIRAP-4x ; S. Akira (Osaka University) and L. O'Neill (Trinity College, Dublin) for TLR4-, MyD88-, TRIF- or TIRAP-deficient mice; L. Stephens (Babraham Institute) for SHIP-1-deficient mice; L. Baud (Hôpital Tenon) for mice with transgenic expression of calpastatin; G. Schiavo (London Cancer Research Institute) for the 2C11 mouse monoclonal antibody to PtdIns(4,5)P2; T. Kinashi (Kyoto University) for the 5′ Myc membrane-targeted version of p110δ in the pMX-neo vector; T. Maffucci (Queen Mary, University of London) for GST–PLC-δ-PH and GST–Akt-PH; H. Stenmark (Oslo University Hospital) for GST–Hrs-FYVE; D. Gray (University of Edinburgh) for the XL-60 cell line; and B. Manoury, N. Leslie and members of the Centre for Cell Signalling for comments. Supported by the European Union Marie Curie (IEF-041713 to E.A. and IEF-274749 to S.T.); the European Molecular Biology Organization (ALTF 1083-2007 to E.A.); the Fondation ARC pour la Recherche sur la Cancer (SAE P2009); the Fund for Scientific Research Flanders; the Hercules Foundation and University of Ghent Multidisciplinary Research Partnership (for work in the laboratory of R.B.); the European Union Marie Curie International Graduate Program in Molecular Medicine (M.A.W.); and Cancer Research UK (C23338/A10200), the Ludwig Institute for Cancer Research and Queen Mary University of London (for work in the laboratory of B.V.).

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Author notes

  1. David Torres and Sandrine Delbauve: These authors contributed equally to this work.

Authors and Affiliations

  1. Centre for Cell Signaling, Barts Institute of Cancer, Queen Mary, University of London, London, UK

    Ezra Aksoy, Salma Taboubi, Maria A Whitehead, Wayne P Pearce, Inma M Berenjeno, Gemma Nock & Bart Vanhaesebroeck

  2. Institute for Medical Immunology, Free University of Brussels, Gosselies, Belgium

    David Torres, Sandrine Delbauve & Veronique Flamand

  3. Division of Cell and Molecular Biology, Centre for Molecular Microbiology and Infection, Imperial College London, London, UK

    Abderrahman Hachani & Alain Filloux

  4. Department for Molecular Biomedical Research, Unit of Molecular Signal Transduction in Inflammation, VIB, Ghent, Belgium

    Rudi Beyaert

  5. Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium

    Rudi Beyaert

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Contributions

E.A. and B.V. developed the main hypothesis; E.A., S.T., D.T., S.D., A.H., W.P.P., I.M.B., G.N. and V.F. planned studies, did experiments and/or analyzed data; M.A.W., A.F. and R.B. contributed reagents, intellectual input and editorial assistance; E.A. and B.V. wrote the paper; B.V. supervised the project; and E.A. and B.V. obtained funding.

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Correspondence to Ezra Aksoy or Bart Vanhaesebroeck.

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Competing interests

B.V. is an advisor to Intellikine, GlaxoSmithKline and Activiomics.

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Aksoy, E., Taboubi, S., Torres, D. et al. The p110δ isoform of the kinase PI(3)K controls the subcellular compartmentalization of TLR4 signaling and protects from endotoxic shock. Nat Immunol 13, 1045–1054 (2012). https://doi.org/10.1038/ni.2426

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