Vif hijacks CBF-β to degrade APOBEC3G and promote HIV-1 infection (original) (raw)
- Letter
- Published: 21 December 2011
- Dong Young Kim3 na1,
- Judd F. Hultquist4 na1,
- Keisuke Shindo4,
- Rebecca S. LaRue4,
- Eunju Kwon3,
- Ming Li4,
- Brett D. Anderson4,
- Linda Yen3,
- David Stanley3,
- Cathal Mahon1,2,3,
- Joshua Kane1,2,
- Kathy Franks-Skiba1,2,
- Peter Cimermancic2,5,
- Alma Burlingame2,3,
- Andrej Sali2,3,5,6,
- Charles S. Craik2,3,
- Reuben S. Harris4,
- John D. Gross2,3,6 &
- …
- Nevan J. Krogan1,2,6,7
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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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of the Krogan, Gross and Harris laboratories for comments, J. R. Johnson for mass spectrometry, B. Leonard for sharing unpublished data, M. Shales for help with figures, and B. Chesebro, D. Gabuzda, J. Lingappa, M. Malim, K. Wehrly, X. F. Yu, A. Bullock, B. Schulman and the AIDS Research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
- Stefanie Jäger, Dong Young Kim and Judd F. Hultquist: These authors contributed equally to this work.
Authors and Affiliations
- 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 - 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 - 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 - 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 - Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, 94158, California, USA
Peter Cimermancic & Andrej Sali - HPC (Host Pathogen Circuitry) Group, University of California-San Francisco, San Francisco, 94158, California, USA
Andrej Sali, John D. Gross & Nevan J. Krogan - J. David Gladstone Institutes, San Francisco, 94158, California, USA
Nevan J. Krogan
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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.
Corresponding authors
Correspondence toReuben S. Harris, John D. Gross or Nevan J. Krogan.
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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
- Received: 26 March 2011
- Accepted: 01 November 2011
- Published: 21 December 2011
- Issue Date: 19 January 2012
- DOI: https://doi.org/10.1038/nature10693
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Editorial Summary
Vif–CBF-β interaction an anti-HIV-1 target
The transcription cofactor CBF-β (core binding factor β) regulates the DNA binding activity of RUNX family proteins. Two independent studies now show that CBF-β also regulates the ability of HIV-1 to evade host restriction mediated by the cDNA deaminase APOBEC3G, a host factor that blocks viral replication. They show that it associates with the HIV protein Vif, and is essential for the assembly of the Vif-Cul5 E3 ubiquitin ligase complex, which mediates the ubiquitination and destruction of APOBEC3. Both groups suggest that disrupting the Vif–CBF-β interaction could provide a new therapeutic target against HIV-1 infection.