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MX2 is an interferon-induced inhibitor of HIV-1 infection

Abstract

HIV-1 replication can be inhibited by type I interferon (IFN), and the expression of a number of gene products with anti-HIV-1 activity is induced by type I IFN1,2. However, none of the known antiretroviral proteins can account for the ability of type I IFN to inhibit early, preintegration phases of the HIV-1 replication cycle in human cells3,4. Here, by comparing gene expression profiles in cell lines that differ in their ability to support the inhibitory action of IFN-α at early steps of the HIV-1 replication cycle, we identify myxovirus resistance 2 (MX2) as an interferon-induced inhibitor of HIV-1 infection. Expression of MX2 reduces permissiveness to a variety of lentiviruses, whereas depletion of MX2 using RNA interference reduces the anti-HIV-1 potency of IFN-α. HIV-1 reverse transcription proceeds normally in MX2-expressing cells, but 2-long terminal repeat circular forms of HIV-1 DNA are less abundant, suggesting that MX2 inhibits HIV-1 nuclear import, or destabilizes nuclear HIV-1 DNA. Consistent with this notion, mutations in the HIV-1 capsid protein that are known, or suspected, to alter the nuclear import pathways used by HIV-1 confer resistance to MX2, whereas preventing cell division increases MX2 potency. Overall, these findings indicate that MX2 is an effector of the anti-HIV-1 activity of type-I IFN, and suggest that MX2 inhibits HIV-1 infection by inhibiting capsid-dependent nuclear import of subviral complexes.

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Figure 1: Differential effects of IFN-α on HIV-1 infection of monocytoid cell lines correlates with MX2 expression.
Figure 2: Inhibition of lentivirus infection by wild-type and mutant MX2, but not other differentially interferon-induced genes.
Figure 3: MX2 inhibits replication-competent HIV-1 and is required for the full antiviral activity of IFN-α.
Figure 4: MX2 activity reduces levels of nuclear HIV-1 DNA, is capsid dependent and is more potent in non-dividing cells.

References

  1. Ho, D. D. et al. Recombinant human interferon alfa-A suppresses HTLV-III replication in vitro. Lancet 325, 602–604 (1985)

    Article  Google Scholar 

  2. Neil, S. & Bieniasz, P. Human immunodeficiency virus, restriction factors, and interferon. J. Interferon Cytokine Res. 29, 569–580 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Bitzegeio, J., Sampias, M., Bieniasz, P. D. & Hatziioannou, T. Adaptation to the interferon-induced antiviral state by human and simian immunodeficiency viruses. J. Virol. 87, 3549–3560 (2013)

    Article  CAS  Google Scholar 

  4. Goujon, C. et al. Evidence for IFNα-induced, SAMHD1-independent inhibitors of early HIV-1 infection. Retrovirology 10, 23 (2013)

    Article  CAS  Google Scholar 

  5. Sheehy, A. M., Gaddis, N. C., Choi, J. D. & Malim, M. H. Isolation of a human gene that inhibits HIV-1 infection and is suppressed by the viral Vif protein. Nature 418, 646–650 (2002)

    Article  ADS  CAS  Google Scholar 

  6. Neil, S. J., Zang, T. & Bieniasz, P. D. Tetherin inhibits retrovirus release and is antagonized by HIV-1 Vpu. Nature 451, 425–430 (2008)

    Article  ADS  CAS  Google Scholar 

  7. Haller, O., Staeheli, P. & Kochs, G. Interferon-induced Mx proteins in antiviral host defense. Biochimie 89, 812–818 (2007)

    Article  CAS  Google Scholar 

  8. Schoggins, J. W. et al. A diverse range of gene products are effectors of the type I interferon antiviral response. Nature 472, 481–485 (2011)

    Article  ADS  CAS  Google Scholar 

  9. Stremlau, M. et al. The cytoplasmic body component TRIM5α restricts HIV-1 infection in Old World monkeys. Nature 427, 848–853 (2004)

    Article  ADS  CAS  Google Scholar 

  10. Melén, K. et al. Human MxB protein, an interferon-α-inducible GTPase, contains a nuclear targeting signal and is localized in the heterochromatin region beneath the nuclear envelope. J. Biol. Chem. 271, 23478–23486 (1996)

    Article  Google Scholar 

  11. King, M. C., Raposo, G. & Lemmon, M. A. Inhibition of nuclear import and cell-cycle progression by mutated forms of the dynamin-like GTPase MxB. Proc. Natl Acad. Sci. USA 101, 8957–8962 (2004)

    Article  ADS  CAS  Google Scholar 

  12. Goujon, C. & Malim, M. H. Characterization of the alpha interferon-induced postentry block to HIV-1 infection in primary human macrophages and T cells. J. Virol. 84, 9254–9266 (2010)

    Article  CAS  Google Scholar 

  13. Yamashita, M., Perez, O., Hope, T. J. & Emerman, M. Evidence for direct involvement of the capsid protein in HIV infection of nondividing cells. PLoS Pathog. 3, e156 (2007)

    Article  Google Scholar 

  14. Yamashita, M. & Emerman, M. Capsid is a dominant determinant of retrovirus infectivity in nondividing cells. J. Virol. 78, 5670–5678 (2004)

    Article  CAS  Google Scholar 

  15. Dismuke, D. J. & Aiken, C. Evidence for a functional link between uncoating of the human immunodeficiency virus type 1 core and nuclear import of the viral preintegration complex. J. Virol. 80, 3712–3720 (2006)

    Article  CAS  Google Scholar 

  16. Lee, K. et al. Flexible use of nuclear import pathways by HIV-1. Cell Host Microbe 7, 221–233 (2010)

    Article  CAS  Google Scholar 

  17. Schaller, T. et al. HIV-1 capsid-cyclophilin interactions determine nuclear import pathway, integration targeting and replication efficiency. PLoS Pathog. 7, e1002439 (2011)

    Article  CAS  Google Scholar 

  18. Koh, Y. et al. Differential effects of human immunodeficiency virus type 1 capsid and cellular factors nucleoporin 153 and LEDGF/p75 on the efficiency and specificity of viral DNA integration. J. Virol. 87, 648–658 (2013)

    Article  CAS  Google Scholar 

  19. Rihn, S. J. et al. Extreme genetic fragility of the HIV-1 capsid. PLoS Pathog. 9, e1003461 (2013)

    Article  CAS  Google Scholar 

  20. Sokolskaja, E., Sayah, D. M. & Luban, J. Target cell cyclophilin A modulates human immunodeficiency virus type 1 infectivity. J. Virol. 78, 12800–12808 (2004)

    Article  CAS  Google Scholar 

  21. Stoye, J. P. Fv1, the mouse retrovirus resistance gene. Rev. Sci. Tech. 17, 269–277 (1998)

    Article  CAS  Google Scholar 

  22. Yamashita, M. & Emerman, M. Cellular restriction targeting viral capsids perturbs human immunodeficiency virus type 1 infection of nondividing cells. J. Virol. 83, 9835–9843 (2009)

    Article  CAS  Google Scholar 

  23. De Iaco, A. et al. TNPO3 protects HIV-1 replication from CPSF6-mediated capsid stabilization in the host cell cytoplasm. Retrovirology 10, 20 (2013)

    Article  CAS  Google Scholar 

  24. Bainbridge, J. W. et al. In vivo gene transfer to the mouse eye using an HIV-based lentiviral vector; efficient long-term transduction of corneal endothelium and retinal pigment epithelium. Gene Ther. 8, 1665–1668 (2001)

    Article  CAS  Google Scholar 

  25. Hatziioannou, T., Cowan, S., Von Schwedler, U. K., Sundquist, W. I. & Bieniasz, P. D. Species-specific tropism determinants in the human immunodeficiency virus type 1 capsid. J. Virol. 78, 6005–6012 (2004)

    Article  CAS  Google Scholar 

  26. Hatziioannou, T., Cowan, S., Goff, S. P., Bieniasz, P. D. & Towers, G. J. Restriction of multiple divergent retroviruses by Lv1 and Ref1. EMBO J. 22, 385–394 (2003)

    Article  CAS  Google Scholar 

  27. Mitrophanous, K. et al. Stable gene transfer to the nervous system using a non-primate lentiviral vector. Gene Ther. 6, 1808–1818 (1999)

    Article  CAS  Google Scholar 

  28. Kemler, I., Barraza, R. & Poeschla, E. M. Mapping the encapsidation determinants of feline immunodeficiency virus. J. Virol. 76, 11889–11903 (2002)

    Article  CAS  Google Scholar 

  29. Ochsenbauer, C. et al. Generation of transmitted/founder HIV-1 infectious molecular clones and characterization of their replication capacity in CD4 T lymphocytes and monocyte-derived macrophages. J. Virol. 86, 2715–2728 (2012)

    Article  CAS  Google Scholar 

  30. Jouvenet, N. et al. Plasma membrane is the site of productive HIV-1 particle assembly. PLoS Biol. 4, e435 (2006)

    Article  Google Scholar 

Download references

Acknowledgements

We thank members of The Rockefeller University Genomics Resource Center for assistance with the microarray experiments and members of the Bieniasz laboratory for discussion and advice. This work was supported by grants from the National Institutes of Health; R37AI64003 (to P.D.B.), R01AI078788 (to T.H.) R01AI100720 (to M.Y.), AI091707 to C.M.R., AI057158 (to I. Lipkin, Northeast Biodefense Center, subcontracted to C.M.R.) and DK095031 to J.W.S., the Greenberg Medical Research Institute and the Starr Foundation (C.M.R.) and by the Howard Hughes Medical Institute.

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Authors and Affiliations

Authors

Contributions

M.K., S.S.Y., J.B., S.B.K., T.Z. and S.J.W. designed and executed the experiments and analysed the data. J.W.S. and C.M.R. provided an interferon-stimulated gene library and advice. M.Y. provided reagents and advice. T.H. provided reagents and advice and supervised the work. P.D.B. conceived the study, supervised the work and wrote the paper, with additional input from all authors.

Corresponding author

Correspondence to Paul D. Bieniasz.

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The authors declare no competing financial interests.

Extended data figures and tables

Extended Data Figure 1 Candidate anti-HIV-1 genes from the microarray analysis.

mRNA levels, determined using Illumina BeadChips and given in arbitrary units, for genes whose differential induction in undifferentiated and PMA-treated THP-1, K562 and U937 cells correlated best with the anti-HIV-1 effect of IFN-α.

Extended Data Figure 2 Additional candidate anti-HIV-1 genes from the microarray analysis.

mRNA levels, determined using Illumina BeadChips and given in arbitrary units, for genes whose differential induction in undifferentiated and MA-treated THP-1, K562 and U937 cells correlated to some degree with the anti-HIV-1 effect of IFN-α.

Extended Data Figure 3 Induction of MX2 by IFN-α in primary CD4+ T cells and macrophages.

Western blot analysis of MX2 and tubulin expression in purified CD4+ T cells, activated with PHA or anti-CD3/CD28, and macrophages treated for 24 h with the indicated doses of IFN-α. Numbers below each lane indicate fluorescence intensity associated with the MX2 band. The second more rapidly migrating MX2 species that was detected inconsistently is of unknown provenance, and may represent a proteolytic breakdown product, or may arise through the use of an alternative start codon at amino acid 25, generating an MX2 protein that lacks the NLS.

Extended Data Figure 4 MX2 is required for the full antiviral activity of IFN-α in HOS cells.

a, Western blot analysis of MX2 expression HOS cells transduced with vectors expressing control or MX2-targeted shRNAs, and treated with IFN-α. Numbers below each lane indicate fluorescence intensity associated with the MX2 band. b, Infectious titre of an HIV-1–GFP reporter virus determined using the shRNA-containing HOS cells from a, with or without IFN-α treatment. Titres are mean + s.d., n = 3 technical replicates, P values calculated using unpaired t-test, representative of three experiments.

Extended Data Figure 5 MX2 activity reduces levels of nuclear HIV-1 DNA in K562 cells.

Quantitative PCR analysis of reverse transcript (left) and 2-LTR circle (right) abundance in empty-vector untreated (none) nevirapine-treated or MX2-expressing K562 cells.

Extended Data Figure 6 Localization of MX2 at nuclear pores.

a, Deconvolution microscopic images (single optical sections) of immunofluorescently stained NUP98 (red), haemagglutinin (HA)-tagged MX2 (green, expressed using CSIB vectors) and DAPI (4′,6-diamidino-2-phenylindole)-stained DNA (blue) in HOS cells. The top set of panels is an optical section approximately through the centre of the vertical dimension of the nucleus, whereas the middle and bottom panels are an optical section approximately coincident with the dorsal surface of the nucleus. The bottom panels are an expanded view of a portion of the centre panels. Scale bars, 10 μm (top), 5 μm (middle) and 1 μm (bottom). b, Pearson’s coefficient for colocalization of MX1 or MX2 and NUP98. Each data point represents an individual cell and the horizontal bar is the mean (n = 6 for MX1, n = 10 for MX2).

Extended Data Figure 7 The N57S capsid mutation reduces HIV-1 sensitivity to IFN-α in HOS cells.

Infectivity of wild-type and N57S CA-mutant HIV-1–GFP reporter viruses in untreated and IFN-α-treated HOS cells. Titres are mean + s.d, n = 3 technical replicates, representative of three experiments. Fold inhibition is the ratio of the titres on untreated and IFN-α-treated cells.

Extended Data Figure 8 Effect of MX2 on HIV-1 and murine leukaemia virus infection in dividing and non-dividing cells.

a, MLV–GFP reporter virus infection of dividing and non-dividing (aphidicolin-treated) vector or MX2-expressing HOS cell clones. b, HIV-1–GFP reporter virus infection of dividing and non-dividing (aphidicolin-treated) vector or MX2-expressing K562 cell clones. c, MLV–GFP reporter virus infection of dividing and non-dividing (aphidicolin-treated) vector or MX2-expressing K562 cell clones.

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Kane, M., Yadav, S., Bitzegeio, J. et al. MX2 is an interferon-induced inhibitor of HIV-1 infection. Nature 502, 563–566 (2013). https://doi.org/10.1038/nature12653

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