Abstract

To evade host immune defences, human immunodeficiency viruses 1 and 2 (HIV-1 and HIV-2) have evolved auxiliary proteins that target cell restriction factors. Viral protein X (Vpx) from the HIV-2/SIVsmm lineage enhances viral infection by antagonizing SAMHD1 (refs 1,2), but this antagonism is not sufficient to explain all Vpx phenotypes. Here, through a proteomic screen, we identified another Vpx target—HUSH (TASOR, MPP8 and periphilin)—a complex involved in position-effect variegation3. HUSH downregulation by Vpx is observed in primary cells and HIV-2-infected cells. Vpx binds HUSH and induces its proteasomal degradation through the recruitment of the DCAF1 ubiquitin ligase adaptor, independently from SAMHD1 antagonism. As a consequence, Vpx is able to reactivate HIV latent proviruses, unlike Vpx mutants, which are unable to induce HUSH degradation. Although antagonism of human HUSH is not conserved among all lentiviral lineages including HIV-1, it is a feature of viral protein R (Vpr) from simian immunodeficiency viruses (SIVs) of African green monkeys and from the divergent SIV of l’Hoest's monkey, arguing in favour of an ancient lentiviral species-specific vpx/vpr gene function. Altogether, our results suggest the HUSH complex as a restriction factor, active in primary CD4+ T cells and counteracted by Vpx, therefore providing a molecular link between intrinsic immunity and epigenetic control.

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Acknowledgements

The authors thank C. Pique and her group and M. Mangeney for discussions, S. Emiliani and A. Abdouni for their advice and protocols for conducting the ChIP experiment, L. Guéguen and L. Picard for the Bio++ analyses and their input on evolutionary analyses, S. Jacquet for her comments on the manuscript, and A. Cimarelli, head of the ‘Host–Pathogen Interaction during Lentiviral Infection’ team at CIRI Lyon. The authors also thank M. Mangeney, G. Lê-Bury and F. Niedergang for their frequent gifts of primary cells, E. Verdin for J-Lat clones, A. Adachi and N. Manel for HIV-2 constructs, N. Landau for SIV packaging constructs and A. Cimarelli for HIV-1- and SIV-based transfer vectors, M.-L. Blondot for her contribution to the SILAC screen and E. Le Rouzic for raising some Vpx mutants. The authors acknowledge the Cytometry and Immunobiology Facility of the Cochin Institute and the 3P5 proteomic facility of Paris Descartes University. The authors also thank all the contributors of publically available genome sequences. This work was supported by grants from the Agence Nationale de la Recherche sur le SIDA et les hépatites virales (ANRS), SIDACTION, Fondation de France and Fondation pour la Recherche Médicale (FRM, grant no. DEQ20140329528 to F.M.-G.). G.C. acknowedges a fellowship from the French government, S.M.-M. received a fellowship from the Fondation pour la Recherche Médicale (FRM, grant no. DEQ20140329528 to F.M.-G.) and H.L. from SIDACTION. L.E. is supported by the CNRS and by grants from the amfAR (Mathilde Krim Phase II Fellowship no. 109140-58-RKHF), the Fondation pour la Recherche Médicale (FRM ‘Projet Innovant’ no. ING20160435028), the FINOVI (‘Recently Settled Scientist’ grant), the ANRS (no. ECTZ19143) and the ANR LABEX ECOFECT (ANR-11-LABX-0048 of Université de Lyon, within the programme ‘Investissements d’Avenir’ (ANR-11-IDEX-0007) operated by the French National Research Agency).

Author information

Author notes

  1. These authors contributed equally: Roy Matkovic, Michaël M Martin, Marina Morel.

Affiliations

  1. Inserm, U1016, Institut Cochin, Paris, France

    • Ghina Chougui
    • , Soundasse Munir-Matloob
    • , Roy Matkovic
    • , Michaël M Martin
    • , Marina Morel
    • , Hichem Lahouassa
    • , Marjorie Leduc
    • , Bertha Cecilia Ramirez
    •  & Florence Margottin-Goguet
  2. CNRS, UMR8104, Paris, France

    • Ghina Chougui
    • , Soundasse Munir-Matloob
    • , Roy Matkovic
    • , Michaël M Martin
    • , Marina Morel
    • , Hichem Lahouassa
    • , Marjorie Leduc
    • , Bertha Cecilia Ramirez
    •  & Florence Margottin-Goguet
  3. Université Paris Descartes, Sorbonne Paris Cité, Paris, France

    • Ghina Chougui
    • , Soundasse Munir-Matloob
    • , Roy Matkovic
    • , Michaël M Martin
    • , Marina Morel
    • , Hichem Lahouassa
    • , Marjorie Leduc
    • , Bertha Cecilia Ramirez
    •  & Florence Margottin-Goguet
  4. 3P5 Proteomic Facility of Université Paris Descartes , Paris, France

    • Marjorie Leduc
  5. CIRI – International Center for Infectiology Research, Inserm U1111, Lyon, France

    • Lucie Etienne
  6. CNRS UMR5308, Lyon, France

    • Lucie Etienne
  7. Ecole Normale Supérieure de Lyon, Université Claude Bernard Lyon 1, Lyon, France

    • Lucie Etienne

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Contributions

G.C., S.M.-M. and F.M.-G. conceived the study. G.C., S.M.-M., R.M., M.M.M., M.M. and F.M.-G. designed experiments and interpreted data. G.C., S.M.-M., R.M., M.M.M. and M.M. performed experiments. H.L. contributed to the SILAC screen design and performance. M.L. contributed to the proteomic analysis of SILAC samples. B.C.R. contributed to the design and performance of experiments in primary cells. L.E. designed and performed the phylogenetic and evolutionary analyses, and contributed to the design of the evolutionary-guided functional experiments. G.C. and F.M.-G. wrote the paper. All authors reviewed and edited the manuscript.

Competing interests

The authors declare no competing interests.

Corresponding author

Correspondence to Florence Margottin-Goguet.

Supplementary information

  1. Supplementary Information

    Supplementary Figures 1–6, Supplementary Tables 2–4, blots for Supplementary Figures 1–4.

  2. Reporting Summary

  3. Supplementary Table 1

    TASOR (FAM208A) is the second best downregulated protein in presence of Vpx.

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DOI

https://doi.org/10.1038/s41564-018-0179-6