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Allosteric competitive inactivation of hematopoietic CSF-1 signaling by the viral decoy receptor BARF1

Nature Structural & Molecular Biology volume 19, pages 938–947 (2012)Cite this article

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Abstract

Hematopoietic human colony-stimulating factor 1 (hCSF-1) is essential for innate and adaptive immunity against viral and microbial infections and cancer. The human pathogen Epstein-Barr virus secretes the lytic-cycle protein BARF1 that neutralizes hCSF-1 to achieve immunomodulation. Here we show that BARF1 binds the dimer interface of hCSF-1 with picomolar affinity, away from the cognate receptor–binding site, to establish a long-lived complex featuring three hCSF-1 at the periphery of the BARF1 toroid. BARF1 locks dimeric hCSF-1 into an inactive conformation, rendering it unable to signal via its cognate receptor on human monocytes. This reveals a new functional role for hCSF-1 cooperativity in signaling. We propose a new viral strategy paradigm featuring an allosteric decoy receptor of the competitive type, which couples efficient sequestration and inactivation of the host growth factor to abrogate cooperative assembly of the cognate signaling complex.

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Figure 1: Structural studies of the BARF1–hCSF-1 assembly.
Figure 2: BARF1 targets hCSF-1 through a new and limited binding site.
Figure 3: BARF1 sequesters hCSF-1 with ultrahigh affinity.
Figure 4: BARF1 is a decoy receptor for hCSF-1 and blocks maturation of monocytes into macrophages.
Figure 5: BARF1 abrogates formation of a cooperative hCSF-1R–hCSF-1 ternary complex.
Figure 6: BARF1 can target mCSF-1 but does not abrogate a mCSF-1RD1–D5–mCSF-1 signaling complex.
Figure 7: BARF1 does not block activation of mCSF-1R by mCSF-1.

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Acknowledgements

We thank the Swiss Light Source (SLS), the European Synchrotron Radiation Facility (ESRF) and the Deutsches Elektronen-Synchrotron/European Molecular Biology Laboratory (DESY/EMBL) for synchrotron-beam time allocation and the staff of beamlines X06SA and X06DA (SLS), ID-29 and ID14-3 (ESRF) and X33 (DESY/EMBL) for technical support. Access to these synchrotron facilities is supported by the European Commission under the Seventh Framework Programme: Research Infrastructures, grant agreement number 226716. This research project was supported by grants from the Research Foundation Flanders (FWO) (3G064307, G059710 and G0B7912N) and Ghent University (BOF instrument) to S.N.S.; a Bundesministerium für Bildung und Forschung (BMBF) research grant Sync-Life (Contract: 05K10YEA) to D.I.S.; an ANR-MIME-2006 grant to N.T. and W.P.B.; and an FWO-Odysseus grant to B.N.L. J.E. would like to acknowledge the joint 2011 APS/CCP4 School for assistance with crystallographic structure refinement. We thank the SWITCH Laboratory at VIB-Belgium for access to FFF-MALS facilities and assistance during data collection and analysis. We also thank F. Wang (Harvard University, Boston, Massachusetts, USA) for providing cDNA of rhBARF1 and Christian Gorba (EMBL Hamburg, Hamburg, Germany) for help with the NMA refinement. I.G. thanks the EM platform of the Partnership for Structural Biology (Grenoble) for access to EM equipment. J.E., K.V. and B.V. are supported as research fellows of the FWO. P.P. is a Marie Curie (FP7) post-doctoral fellow.

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  1. Nathalie Bracke, Alexander V Shkumatov & Anaïs Bekaert

    Present address: Present addresses: Laboratory of Drug Quality and Registration, Department of Pharmaceutical Analysis, Ghent University, Ghent, Belgium (N.B.); Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA (A.V.S.); Oxyrane, Ghent, Belgium (A.B.).,

Authors and Affiliations

  1. Department of Biochemistry & Microbiology, Unit for Structural Biology, Laboratory for Protein Biochemistry and Biomolecular Engineering, Ghent University, Ghent, Belgium

    Jonathan Elegheert, Nathalie Bracke, Kenneth Verstraete, Anaïs Bekaert, Bjorn Vergauwen & Savvas N Savvides

  2. Department of Molecular Biomedical Research, Laboratory of Immunoregulation and Mucosal Immunology, VIB and Ghent University, Ghent, Belgium

    Philippe Pouliot & Bart N Lambrecht

  3. Unit for Virus Host-Cell Interactions, Université Joseph Fourier, European Molecular Biology Laboratory and Centre National de la Recherche Scientifique, Grenoble, France

    Irina Gutsche, Nicolas Tarbouriech & Wim P Burmeister

  4. Biological Small Angle Scattering Group, European Molecular Biology Laboratory, Hamburg Unit, Deutsches Elektronen-Synchrotron, Hamburg, Germany

    Alexander V Shkumatov & Dmitri I Svergun

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Contributions

J.E. and N.B. expressed and purified recombinant proteins, with contributions from A.B. J.E. carried out all crystallographic studies with contributions from N.B., A.B. and S.N.S. J.E., N.B. and B.V. carried out ITC with contributions from S.N.S. N.T. and W.P.B. carried out preliminary structural studies on independently produced material. J.E., N.B. and B.V. carried out SPR studies with contributions from N.T. and W.P.B. K.V. provided tools and assistance for tissue culture. P.P. and B.N.L. carried out cellular assays. J.E., A.V.S. and D.I.S. carried out SAXS studies. I.G. carried out EM imaging and data analysis. J.E., N.B., B.V. and S.N.S. established the experimental approach and designed experiments. S.N.S. directed the study. J.E. and S.N.S. wrote the manuscript with contributions from all authors.

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Correspondence to Savvas N Savvides.

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Elegheert, J., Bracke, N., Pouliot, P. et al. Allosteric competitive inactivation of hematopoietic CSF-1 signaling by the viral decoy receptor BARF1. Nat Struct Mol Biol 19, 938–947 (2012). https://doi.org/10.1038/nsmb.2367

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