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Primate immunodeficiency virus proteins Vpx and Vpr counteract transcriptional repression of proviruses by the HUSH complex

Nature Microbiology volume 3, pages 1354–1361 (2018)Cite this article

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Abstract

Host factors that silence provirus transcription in CD4+ memory T cells help HIV-1 escape eradication by the host immune system and by antiviral drugs1. These same factors, however, must be overcome for HIV-1 to propagate. Here we show that Vpx and Vpr encoded by diverse primate immunodeficiency viruses activate provirus transcription. Vpx and Vpr are adaptor proteins for the DCAF1-CUL4A/B E3 ubiquitin ligase that degrade SAMHD1 and increase reverse transcription2,3,4. Nonetheless, Vpx and Vpr have effects on reporter gene expression that are not explained by SAMHD1 degradation5,6,7,8. A screen for factors that mimic these effects identified the human silencing hub (HUSH) complex, FAM208A (TASOR/RAP140), MPHOSPH8 (MPP8), PPHLN1 (PERIPHILIN) and MORC29,10,11,12,13. Vpx associated with the HUSH complex and decreased steady-state level of these proteins in a DCAF1/CUL4A/B/proteasome-dependent manner14,15. Replication kinetics of HIV-1 and SIVMAC was accelerated to a similar extent by vpx or FAM208A knockdown. Finally, vpx increased steady-state levels of LINE-1 ORF1p, as previously described for FAM208A disruption11. These results demonstrate that the HUSH complex represses primate immunodeficiency virus transcription, and that, to counteract this restriction, viral Vpx or Vpr proteins degrade the HUSH complex.

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Fig. 1: Diverse primate immunodeficiency virus vpx and vpr orthologues activate provirus transcription, whether delivered before, during or after reporter provirus integration.
Fig. 2: Vpx activates provirus transcription by degrading HUSH complex components.
Fig. 3: The HIV-1 LTR is activated by Vpx or disruption of FAM208A.
Fig. 4: Vpx counteracts FAM208A restriction of exogenous HIV-1 and SIVMAC239 infection and of endogenous LINE-1.

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Data availability

All data needed to evaluate the conclusions in the paper are present in the paper or in the supplementary table. The plasmids described in Supplementary Table 1, along with their complete nucleotide sequences, are available at https://www.addgene.org/Jeremy_Luban/.

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Acknowledgements

The authors wish to dedicate these experiments to the memory of Jan Svoboda (1934–2017), whose demonstration that cells may carry Rous sarcoma virus genetic information in the absence of any infectious virus production provided support to the proviral hypothesis. We thank L. Cavacini for antibodies and N. Landau for SIV plasmids. The following reagents were obtained through the AIDS Reagent Program, Division of AIDS, NIAID, NIH: J-Lat Tat-GFPCells (A1) from Dr. Eric Verdin and SIVmac239 SpX and SIVmac239 SpX ΔVpx from Dr. Ronald C. Desrosiers. This research was supported by National Institutes of Health (USA) grants nos. R01AI111809, RO1AI117839 and DP1DA034990 to J.L.

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  1. Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA

    Leonid Yurkovetskiy, Mehmet Hakan Guney, Kyusik Kim, Shih Lin Goh, Sean McCauley, Ann Dauphin, William E. Diehl & Jeremy Luban

  2. Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA

    Jeremy Luban

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Contributions

L.Y. and J.L. designed the experiments. L.Y. performed the experiments with assistance from M.H.G., K.K., S.L.G., S.M., A.D. and W.E.D. L.Y. and J.L. analysed the experimental data. All authors contributed to the writing of the manuscript.

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Correspondence to Jeremy Luban.

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Supplementary Figures 1–5 and Supplementary Table 1.

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Yurkovetskiy, L., Guney, M.H., Kim, K. et al. Primate immunodeficiency virus proteins Vpx and Vpr counteract transcriptional repression of proviruses by the HUSH complex. Nat Microbiol 3, 1354–1361 (2018). https://doi.org/10.1038/s41564-018-0256-x

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