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Vif hijacks CBF-β to degrade APOBEC3G and promote HIV-1 infection

Nature volume 481, pages 371–375 (2012)Cite this article

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Abstract

Restriction factors, such as the retroviral complementary DNA deaminase APOBEC3G, are cellular proteins that dominantly block virus replication1,2,3. The AIDS virus, human immunodeficiency virus type 1 (HIV-1), produces the accessory factor Vif, which counteracts the host’s antiviral defence by hijacking a ubiquitin ligase complex, containing CUL5, ELOC, ELOB and a RING-box protein, and targeting APOBEC3G for degradation4,5,6,7,8,9,10. Here we reveal, using an affinity tag/purification mass spectrometry approach, that Vif additionally recruits the transcription cofactor CBF-β to this ubiquitin ligase complex. CBF-β, which normally functions in concert with RUNX DNA binding proteins, allows the reconstitution of a recombinant six-protein assembly that elicits specific polyubiquitination activity with APOBEC3G, but not the related deaminase APOBEC3A. Using RNA knockdown and genetic complementation studies, we also demonstrate that CBF-β is required for Vif-mediated degradation of APOBEC3G and therefore for preserving HIV-1 infectivity. Finally, simian immunodeficiency virus (SIV) Vif also binds to and requires CBF-β to degrade rhesus macaque APOBEC3G, indicating functional conservation. Methods of disrupting the CBF-β–Vif interaction might enable HIV-1 restriction and provide a supplement to current antiviral therapies that primarily target viral proteins.

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Figure 1: AP–MS experiments identify
Figure 2: CBF-β is a stoichiometric component of the Vif E3 ubiquitin ligase.
Figure 3: CBF-β and Vif collaborate to degrade APOBEC3G and enable HIV-1 infectivity.
Figure 4: Model for Vif–CBF-β E3 ligase formation and APOBEC3G polyubiquitination and degradation.

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References

  1. Goff, S. P. Retrovirus restriction factors. Mol. Cell 16, 849–859 (2004)

    Article  CAS  Google Scholar 

  2. Malim, M. H. & Emerman, M. HIV-1 accessory proteins—ensuring viral survival in a hostile environment. Cell Host Microbe 3, 388–398 (2008)

    Article  CAS  Google Scholar 

  3. Albin, J. S. & Harris, R. S. Interactions of host APOBEC3 restriction factors with HIV-1 in vivo: implications for therapeutics. Expert Rev. Mol. Med. 12, e4 (2010)

    Article  Google Scholar 

  4. Yu, X. et al. Induction of APOBEC3G ubiquitination and degradation by an HIV-1 Vif-Cul5-SCF complex. Science 302, 1056–1060 (2003)

    Article  ADS  CAS  Google Scholar 

  5. Mehle, A., Goncalves, J., Santa-Marta, M., McPike, M. & Gabuzda, D. Phosphorylation of a novel SOCS-box regulates assembly of the HIV-1 Vif-Cul5 complex that promotes APOBEC3G degradation. Genes Dev. 18, 2861–2866 (2004)

    Article  CAS  Google Scholar 

  6. Mehle, A. et al. Vif overcomes the innate antiviral activity of APOBEC3G by promoting its degradation in the ubiquitin-proteasome pathway. J. Biol. Chem. 279, 7792–7798 (2004)

    Article  CAS  Google Scholar 

  7. 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 

  8. Sheehy, A. M., Gaddis, N. C. & Malim, M. H. The antiretroviral enzyme APOBEC3G is degraded by the proteasome in response to HIV-1 Vif. Nature Med. 9, 1404–1407 (2003)

    Article  CAS  Google Scholar 

  9. Conticello, S. G., Harris, R. S. & Neuberger, M. S. The Vif protein of HIV triggers degradation of the human antiretroviral DNA deaminase APOBEC3G. Curr. Biol. 13, 2009–2013 (2003)

    Article  CAS  Google Scholar 

  10. Stopak, K., de Noronha, C., Yonemoto, W. & Greene, W. C. HIV-1 Vif blocks the antiviral activity of APOBEC3G by impairing both its translation and intracellular stability. Mol. Cell 12, 591–601 (2003)

    Article  CAS  Google Scholar 

  11. Passmore, L. A. & Barford, D. Getting into position: the catalytic mechanisms of protein ubiquitylation. Biochem. J. 379, 513–525 (2004)

    Article  CAS  Google Scholar 

  12. Thrower, J. S., Hoffman, L., Rechsteiner, M. & Pickart, C. M. Recognition of the polyubiquitin proteolytic signal. EMBO J. 19, 94–102 (2000)

    Article  CAS  Google Scholar 

  13. Stanley, B. J. et al. Structural insight into the human immunodeficiency virus Vif SOCS box and its role in human E3 ubiquitin ligase assembly. J. Virol. 82, 8656–8663 (2008)

    Article  CAS  Google Scholar 

  14. Jäger, S. et al. Purification and characterization of HIV-human protein complexes. Methods 53, 13–19 (2011)

    Article  Google Scholar 

  15. Jäger, S. et al. Global landscape of HIV–human protein complexes. Nature doi:10.1038/nature10719 (this issue).

  16. Kamura, T. et al. VHL-box and SOCS-box domains determine binding specificity for Cul2-Rbx1 and Cul5-Rbx2 modules of ubiquitin ligases. Genes Dev. 18, 3055–3065 (2004)

    Article  CAS  Google Scholar 

  17. Huang, D. T. et al. E2-RING expansion of the NEDD8 cascade confers specificity to cullin modification. Mol. Cell 33, 483–495 (2009)

    Article  CAS  Google Scholar 

  18. Wong, W. F., Kohu, K., Chiba, T., Sato, T. & Satake, M. Interplay of transcription factors in T-cell differentiation and function: the role of Runx. Immunology 132, 157–164 (2011)

    Article  CAS  Google Scholar 

  19. Petroski, M. D. & Deshaies, R. J. Mechanism of lysine 48-linked ubiquitin-chain synthesis by the cullin-RING ubiquitin-ligase complex SCF-Cdc34. Cell 123, 1107–1120 (2005)

    Article  CAS  Google Scholar 

  20. Kim, H. T. et al. Certain pairs of ubiquitin-conjugating enzymes (E2s) and ubiquitin-protein ligases (E3s) synthesize nondegradable forked ubiquitin chains containing all possible isopeptide linkages. J. Biol. Chem. 282, 17375–17386 (2007)

    Article  CAS  Google Scholar 

  21. DeHart, J. L., Bosque, A., Harris, R. S. & Planelles, V. Human immunodeficiency virus type 1 Vif induces cell cycle delay via recruitment of the same E3 ubiquitin ligase complex that targets APOBEC3 proteins for degradation. J. Virol. 82, 9265–9272 (2008)

    Article  CAS  Google Scholar 

  22. Stenglein, M. D., Burns, M. B., Li, M., Lengyel, J. & Harris, R. S. APOBEC3 proteins mediate the clearance of foreign DNA from human cells. Nature Struct. Mol. Biol. 17, 222–229 (2010)

    Article  CAS  Google Scholar 

  23. Hultquist, J. F. et al. Human and rhesus APOBEC3D, APOBEC3F, APOBEC3G, and APOBEC3H demonstrate a conserved capacity to restrict Vif-deficient HIV-1. J. Virol. 85, 11220–11234 (2011)

    Article  CAS  Google Scholar 

  24. Rodrigo-Brenni, M. C. & Morgan, D. O. Sequential E2s drive polyubiquitin chain assembly on APC targets. Cell 130, 127–139 (2007)

    Article  CAS  Google Scholar 

  25. Wu, K., Kovacev, J. & Pan, Z. Q. Priming and extending: a UbcH5/Cdc34 E2 handoff mechanism for polyubiquitination on a SCF substrate. Mol. Cell 37, 784–796 (2010)

    Article  CAS  Google Scholar 

  26. Koning, F. A. et al. Defining APOBEC3 expression patterns in human tissues and hematopoietic cell subsets. J. Virol. 83, 9474–9485 (2009)

    Article  CAS  Google Scholar 

  27. Refsland, E. W. et al. Quantitative profiling of the full APOBEC3 mRNA repertoire in lymphocytes and tissues: implications for HIV-1 restriction. Nucleic Acids Res. 38, 4274–4284 (2010)

    Article  CAS  Google Scholar 

  28. Saha, A. & Deshaies, R. J. Multimodal activation of the ubiquitin ligase SCF by Nedd8 conjugation. Mol. Cell 32, 21–31 (2008)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank members of the Krogan, Gross and Harris laboratories for comments, J. R. Johnson for mass spectrometry, B. Leonard for sharing unpublished data, M. Shales for help with figures, and B. Chesebro, D. Gabuzda, J. Lingappa, M. Malim, K. Wehrly, X. F. Yu, A. Bullock, B. Schulman and the AIDS Research and Reference Reagent Program for reagents. This research was funded by grants from QB3 at University of California, San Francisco, and the National Institutes of Health (P50 GM082250, P01 AI090935 and P50 GM081879 to N.J.K.; U54 RR022220 to A.S.; R01 AI064046 and P01 GM091743 to R.S.H.; P50 GM082250 to J.D.G. and C.S.C.; P41RR001614 and P50GM081879 to A.B.). N.J.K. is a Searle Scholar and a Keck Young Investigator.

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Author notes

  1. Stefanie Jäger, Dong Young Kim and Judd F. Hultquist: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cellular and Molecular Pharmacology, University of California-San Francisco, San Francisco, 94158, California, USA

    Stefanie Jäger, Cathal Mahon, Joshua Kane, Kathy Franks-Skiba & Nevan J. Krogan

  2. California Institute for Quantitative Biosciences, QB3, San Francisco, California, California 94158, USA ,

    Stefanie Jäger, Cathal Mahon, Joshua Kane, Kathy Franks-Skiba, Peter Cimermancic, Alma Burlingame, Andrej Sali, Charles S. Craik, John D. Gross & Nevan J. Krogan

  3. Department of Pharmaceutical Chemistry, University of California, San Francisco, 94158, California, USA

    Dong Young Kim, Eunju Kwon, Linda Yen, David Stanley, Cathal Mahon, Alma Burlingame, Andrej Sali, Charles S. Craik & John D. Gross

  4. Department of Biochemistry, Molecular Biology and Biophysics, Institute for Molecular Virology, Center for Genome Engineering, University of Minnesota, Minneapolis, 55455, Minnesota, USA

    Judd F. Hultquist, Keisuke Shindo, Rebecca S. LaRue, Ming Li, Brett D. Anderson & Reuben S. Harris

  5. Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, 94158, California, USA

    Peter Cimermancic & Andrej Sali

  6. HPC (Host Pathogen Circuitry) Group, University of California-San Francisco, San Francisco, 94158, California, USA

    Andrej Sali, John D. Gross & Nevan J. Krogan

  7. J. David Gladstone Institutes, San Francisco, 94158, California, USA

    Nevan J. Krogan

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  1. Stefanie Jäger
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Contributions

K.S., R.S.L. and E.K. made equal secondary contributions to this work. S.J. and K.F.S. generated the Vif protein–protein interaction map (Fig. 1); S.J. performed the co-immunoprecipitation confirmation assays (Supplementary Fig. 1); P.C. developed and implemented the MiST scoring system (Fig. 1c); S.J. and E.K. performed double purification analyses (Fig. 1c, d); C.M. and J.K. performed immunoprecipitation analyses (Fig. 1f, g); D.Y.K., L.Y. and D.S. reconstituted the Vif E3 ligase from recombinant components and performed ubiquitination assays (Fig. 2 and Supplementary Figs 2–4); M.L. expressed and purified A3 proteins and did in vitro pulldowns (Fig. 2); K.S., J.F.H. and R.S.L. performed CBF-β knockdown, complementation and virus infectivity experiments (Fig. 3 and Supplementary Figs 5–7); and B.D.A. and J.F.H. did the CBF-β–Vif co-immunoprecipitation experiments (Supplementary Fig. 8). A.B., A.S., C.S.C., R.S.H., J.D.G. and N.J.K. supervised the research; S.J., D.Y.K., J.F.H., K.S., R.S.H., J.D.G. and N.J.K. wrote and revised the manuscript.

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Correspondence to Reuben S. Harris, John D. Gross or Nevan J. Krogan.

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The authors declare no competing financial interests.

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Jäger, S., Kim, D., Hultquist, J. et al. Vif hijacks CBF-β to degrade APOBEC3G and promote HIV-1 infection. Nature 481, 371–375 (2012). https://doi.org/10.1038/nature10693

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