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The interferon-inducible isoform of NCOA7 inhibits endosome-mediated viral entry

Nature Microbiology volume 3, pages 1369–1376 (2018)Cite this article

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An Author Correction to this article was published on 22 January 2019

This article has been updated

Abstract

Interferons (IFNs) mediate cellular defence against viral pathogens by upregulation of IFN-stimulated genes whose products interact with viral components or alter cellular physiology to suppress viral replication1,2,3. Among the IFN-stimulated genes that can inhibit influenza A virus (IAV)4 are the myxovirus resistance 1 GTPase5 and IFN-induced transmembrane protein 3 (refs 6,7). Here, we use ectopic expression and gene knockout to demonstrate that the IFN-inducible 219-amino acid short isoform of human nuclear receptor coactivator 7 (NCOA7) is an inhibitor of IAV as well as other viruses that enter the cell by endocytosis, including hepatitis C virus. NCOA7 interacts with the vacuolar H+-ATPase (V-ATPase) and its expression promotes cytoplasmic vesicle acidification, lysosomal protease activity and the degradation of endocytosed antigen. Step-wise dissection of the IAV entry pathway demonstrates that NCOA7 inhibits fusion of the viral and endosomal membranes and subsequent nuclear translocation of viral ribonucleoproteins. Therefore, NCOA7 provides a mechanism for immune regulation of endolysosomal physiology that not only suppresses viral entry into the cytosol from this compartment but may also regulate other V-ATPase-associated cellular processes, such as physiological adjustments to nutritional status, or the maturation and function of antigen-presenting cells.

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Fig. 1: NCOA7 inhibits infection by viruses entering through the endocytic pathway.
Fig. 2: NCOA7 function inhibits IAV membrane fusion.
Fig. 3: NCOA7 interacts with V-ATPase and promotes cytoplasmic vesicle acidification and lysosomal protease activation.

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Data availability

The datasets generated and/or analysed during the current study are available from the corresponding authors on reasonable request.

Change history

  • 22 January 2019

    In the version of Supplementary Fig. 5a originally published with this Letter, the authors mistakenly duplicated images of LAMP1 staining in place of CD63 staining; this has now been amended to the correct version shown below.

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Acknowledgements

We thank W. Barclay, F. Blanchet, M. Bonazzi, L. Espert, M. Huttunen, J. Long, E. Martinez, J. Mercer, D. Muriaux, P. Rocha, R. Schulz and Y. Yamauchi for generous provision of reagents and helpful discussions. We are grateful to A. Vaughan for instruction in IAV amplification, P. J. Chana (Biomedical Research Centre (BRC) Flow Core) for training and advice on imaging flow cytometry, I. Ali (King’s College London Nikon Imaging Centre) for advice with confocal microscopy and image acquisition, I. Parham for help with the time-lapse microscopy, and M. Boyer and B. Monterroso (‘MRI’ imaging and flow cytometry facility) for cell sorting and advice with flow cytometry and confocal microscopy, respectively. This work was supported by the UK Medical Research Council (G1000196 to M.H.M.), the Wellcome Trust (106223/Z/14/Z to M.H.M.), the NIH (DA033773), a Wellcome Trust Research Training Fellowship and National Institute for Health Research BRC King’s Prize Fellowship (to T.D.), the Institut National de la Santé et de la Recherche Médicale (to C.G.), the European Research Council under the European Union’s Horizon 2020 research and innovation programme (grant agreement number 759226 to C.G.), the ATIP-Avenir Program (to C.G.), institutional funds from the Centre National de la Recherche Scientifique and Montpellier University (to C.G.), France Recherche Nord & Sud Sida/HIV et Hépatites (to O.M.), a PhD studentship from the Ministry of Higher Education and Research (to B.B.), King’s College London departmental start-up funds (to M.-T.C.), and the Department of Health via a National Institute for Health Research BRC award to Guy’s and St. Thomas’ NHS Foundation Trust in partnership with King’s College London and King’s College Hospital NHS Foundation Trust. We acknowledge the imaging facility MRI, a member of the national infrastructure France-BioImaging supported by the French National Research Agency (ANR-10-INBS-04).

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  1. Tomas Doyle

    Present address: GlaxoSmithKline Medicines Research Centre, Stevenage, UK

Authors and Affiliations

  1. Department of Infectious Diseases, School of Immunology and Microbial Sciences, King’s College London, London, UK

    Tomas Doyle, Darja Pollpeter, Marion Lussignol, Luis Apolonia, Maria-Teresa Catanese & Michael H. Malim

  2. IRIM, CNRS, Université de Montpellier, Montpellier, France

    Olivier Moncorgé, Boris Bonaventure, Marine Tauziet & Caroline Goujon

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Contributions

T.D., O.M., M.-T.C., C.G. and M.H.M. conceived and designed the experiments. T.D., O.M., B.B., D.P., M.L., M.T., L.A. and C.G. performed the experiments. All authors analysed the data. T.D., C.G. and M.H.M. wrote the manuscript with input from all authors.

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Correspondence to Caroline Goujon or Michael H. Malim.

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Supplementary Figures 1–6

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Supplementary Movie 1

NCOA7 inhibits IAV infection. 293T cells constitutively expressing CD8 or NCOA7 were plated in 48-well plates and transfected with a GFP reporter construct responsive to the IAV RNA polymerase. Cells were infected with A/Eng/195/2009 and visualized by time-lapse microscopy. Productive infection is reflected by GFP expression; time post infection is indicated. The experiment was repeated independently three times with similar results.

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Doyle, T., Moncorgé, O., Bonaventure, B. et al. The interferon-inducible isoform of NCOA7 inhibits endosome-mediated viral entry. Nat Microbiol 3, 1369–1376 (2018). https://doi.org/10.1038/s41564-018-0273-9

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