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. Access through your institution Buy or

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HIV-1 VPR COMBATS THE PU.1-DRIVEN ANTIVIRAL RESPONSE IN PRIMARY HUMAN MACROPHAGES Article Open access 29 June 2024 INFLUENZA VIRUS REPURPOSES THE ANTIVIRAL PROTEIN IFIT2 TO PROMOTE

TRANSLATION OF VIRAL MRNAS Article 24 August 2020 MX2-MEDIATED INNATE IMMUNITY AGAINST HIV-1 IS REGULATED BY SERINE PHOSPHORYLATION Article 19 July 2021 REFERENCES * Ho, D. D. et al.

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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. AUTHOR INFORMATION

Author notes * Shalini S. Yadav, Sam J. Wilson & John W. Schoggins Present address: Present addresses: University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard,

Dallas, Texas 75390-9048, USA (J.W.S.); MRC Centre for Virus Research, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of

Glasgow, Glasgow G12 8QQ, UK (S.J.W.); Weill Cornell Medical College, 525 East 68th street, New York, New York 10025, USA (S.S.Y.)., AUTHORS AND AFFILIATIONS * Aaron Diamond AIDS Research

Center, New York, 10016, New York, USA Melissa Kane, Shalini S. Yadav, Julia Bitzegeio, Sebla B. Kutluay, Trinity Zang, Sam J. Wilson, Masahiro Yamashita, Theodora Hatziioannou & Paul D.

Bieniasz * Laboratory of Retrovirology, The Rockefeller University, New York, 10065, New York, USA Melissa Kane, Shalini S. Yadav, Julia Bitzegeio, Sebla B. Kutluay, Trinity Zang, Sam J.

Wilson & Paul D. Bieniasz * Howard Hughes Medical Institute, New York, 10016, New York, USA Shalini S. Yadav, Trinity Zang, Sam J. Wilson & Paul D. Bieniasz * Center for the Study of

Hepatitis C, The Rockefeller University, New York, 10065, New York, USA John W. Schoggins & Charles M. Rice Authors * Melissa Kane View author publications You can also search for this

author inPubMed Google Scholar * Shalini S. Yadav View author publications You can also search for this author inPubMed Google Scholar * Julia Bitzegeio View author publications You can also

search for this author inPubMed Google Scholar * Sebla B. Kutluay View author publications You can also search for this author inPubMed Google Scholar * Trinity Zang View author

publications You can also search for this author inPubMed Google Scholar * Sam J. Wilson View author publications You can also search for this author inPubMed Google Scholar * John W.

Schoggins View author publications You can also search for this author inPubMed Google Scholar * Charles M. Rice View author publications You can also search for this author inPubMed Google

Scholar * Masahiro Yamashita View author publications You can also search for this author inPubMed Google Scholar * Theodora Hatziioannou View author publications You can also search for

this author inPubMed Google Scholar * Paul D. Bieniasz View author publications You can also search for this author inPubMed Google Scholar 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. ETHICS DECLARATIONS COMPETING INTERESTS 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. POWERPOINT SLIDES

POWERPOINT SLIDE FOR FIG. 1 POWERPOINT SLIDE FOR FIG. 2 POWERPOINT SLIDE FOR FIG. 3 POWERPOINT SLIDE FOR FIG. 4 RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS

ARTICLE 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 Download

citation * Received: 23 August 2013 * Accepted: 12 September 2013 * Published: 13 October 2013 * Issue Date: 24 October 2013 * DOI: https://doi.org/10.1038/nature12653 SHARE THIS ARTICLE

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