Article

Targeting the ATP-dependent formation of herpesvirus ribonucleoprotein particle assembly as an antiviral approach

  • Nature Microbiology 2, Article number: 16201 (2016)
  • doi:10.1038/nmicrobiol.2016.201
  • Download Citation
Received:
Accepted:
Published online:

Abstract

Human herpesviruses are responsible for a range of debilitating acute and recurrent diseases, including a number of malignancies. Current treatments are limited to targeting the herpesvirus DNA polymerases, but with emerging viral resistance and little efficacy against the oncogenic herpesviruses, there is an urgent need for new antiviral strategies. Here, we describe a mechanism to inhibit the replication of the oncogenic herpesvirus Kaposi's sarcoma-associated herpesvirus (KSHV), by targeting the ATP-dependent formation of viral ribonucleoprotein particles (vRNPs). We demonstrate that small-molecule inhibitors which selectively inhibit the ATPase activity of the cellular human transcription/export complex (hTREX) protein UAP56 result in effective inhibition of vRNP formation, viral lytic replication and infectious virion production. Strikingly, as all human herpesviruses use conserved mRNA processing pathways involving hTREX components, we demonstrate the feasibility of this approach for pan-herpesvirus inhibition.

  • Subscribe to Nature Microbiology for full access:

    $59

    Subscribe

  • Purchase article full text and PDF:

    $32

    Buy now

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

Change history

  • Corrected online 14 July 2017

    In the PDF version of this article previously published, the year of publication provided in the footer of each page and in the 'How to cite' section was erroneously given as 2017, it should have been 2016. This error has now been corrected. The HTML version of the article was not affected.

References

  1. 1.

    , , & Herpesvirus infections of the nervous system. Nat. Clin. Pract. Neurol. 3, 82–94 (2007).

  2. 2.

    et al. Utilising proteomic approaches to understand oncogenic human herpesviruses (Review). Mol. Clin. Oncol. 2, 891–903 (2014).

  3. 3.

    et al. β-HHVs and HHV-8 in lymphoproliferative disorders. Mediterr. J. Hematol. Infect. Dis. 3, e2011043 (2011).

  4. 4.

    KSHV infection and the pathogenesis of Kaposi's sarcoma. Annu. Rev. Pathol. 1, 273–296 (2006).

  5. 5.

    , & Kaposi's sarcoma and its associated herpesvirus. Nat. Rev. Cancer 10, 707–719 (2010).

  6. 6.

    , , , & Human herpesvirus KSHV encodes a constitutively active G-protein-coupled receptor linked to cell proliferation. Nature 385, 347–350 (1997).

  7. 7.

    et al. Kaposi's sarcoma-associated herpesvirus-encoded interleukin-6 and G-protein-coupled receptor regulate angiopoietin-2 expression in lymphatic endothelial cells. Cancer Res. 67, 4042–4051 (2007).

  8. 8.

    et al. Kaposi's sarcoma-associated human herpesvirus-8 encodes homologues of macrophage inflammatory protein-1 and interleukin-6. Nat. Med. 3, 287–292 (1997).

  9. 9.

    & The K1 protein of Kaposi's sarcoma-associated herpesvirus activates the Akt signaling pathway. J. Virol. 78, 1918–1927 (2004).

  10. 10.

    et al. Activation of mitogen-activated protein kinase and NF-κB pathways by a Kaposi's sarcoma-associated herpesvirus K15 membrane protein. J. Virol. 77, 9346–9358 (2003).

  11. 11.

    , & A novel mechanism inducing genome instability in Kaposi's sarcoma-associated herpesvirus infected cells. PLoS Pathogens 10, e1004098 (2014).

  12. 12.

    , , , & Herpes simplex virus resistance to acyclovir and penciclovir after two decades of antiviral therapy. Clin. Microbiol. Rev. 16, 114–128 (2003).

  13. 13.

    et al. Cidofovir for the treatment of Kaposi's sarcoma in an HIV-negative homosexual man. Br. J. Dermatol. 141, 1136–1152 (1999).

  14. 14.

    et al. Efficacy of cidofovir on human herpesvirus 8 viraemia and Kaposi's sarcoma progression in two patients with AIDS. AIDS 15, 2061–2062 (2001).

  15. 15.

    et al. A pilot study of cidofovir in patients with Kaposi sarcoma. J. Infect. Dis. 187, 149–153 (2003).

  16. 16.

    et al. Oral ganciclovir for patients with cytomegalovirus retinitis treated with a ganciclovir implant. N. Engl. J. Med. 340, 1063–1070 (1999).

  17. 17.

    et al. Use of antiherpes drugs and the risk of Kaposi's sarcoma: data from the multicenter AIDS cohort study. J. Infect. Dis. 173, 1477–1480 (1996).

  18. 18.

    , , & The DEAD-box protein family of RNA helicases. Gene 367, 17–37 (2006).

  19. 19.

    et al. Discovering new medicines targeting helicases: challenges and recent progress. J. Biomol. Screen. 18, 761–781 (2013).

  20. 20.

    et al. Structure-based discovery of triphenylmethane derivatives as inhibitors of hepatitis C virus helicase. J. Med. Chem. 52, 2716–2723 (2009).

  21. 21.

    , , , & Abrogation of hepatitis C virus NS3 helicase enzymatic activity by recombinant human antibodies. J. Gen. Virol. 84, 2323–2332 (2003).

  22. 22.

    et al. Isolation of specific and high-affinity RNA aptamers against NS3 helicase domain of hepatitis C virus. RNA 10, 1277–1290 (2004).

  23. 23.

    et al. Selective pharmacological targeting of a DEAD Box RNA helicase. PLoS ONE 3, e1583 (2008).

  24. 24.

    et al. Effects of hippuristanol, an inhibitor of eIF4A, on adult T-cell leukemia. Biochem. Pharmacol. 81, 713–722 (2011).

  25. 25.

    et al. Ring expanded nucleoside analogues inhibit RNA helicase and intracellular human immunodeficiency virus type 1 replication. J. Med. Chem. 51, 5043–5051 (2008).

  26. 26.

    & γ-2 Herpes virus post-transcriptional gene regulation. Clin. Microbiol. Infect. 12, 110–117 (2006).

  27. 27.

    , , & Kaposi's sarcoma-associated herpesvirus ORF57 protein: exploiting all stages of viral mRNA processing. Viruses 5, 1901–1923 (2013).

  28. 28.

    , , , & Kaposi's sarcoma-associated herpesvirus ORF57 protein interacts with PYM to enhance translation of viral intronless mRNAs. EMBO J. 29, 1851–1864 (2010).

  29. 29.

    et al. An interaction between KSHV ORF57 and UIF provides mRNA-adaptor redundancy in herpesvirus intronless mRNA export. PLoS Pathogens 7, e1002138 (2011).

  30. 30.

    et al. UIF, a new mRNA export adaptor that works together with REF/ALY, requires FACT for recruitment to mRNA. Curr. Biol. 19, 1918–1924 (2009).

  31. 31.

    et al. ATP is required for interactions between UAP56 and two conserved mRNA export proteins, Aly and CIP29, to assemble the TREX complex. Genes Dev. 24, 2043–2053 (2010).

  32. 32.

    et al. Chtop is a component of the dynamic TREX mRNA export complex. EMBO J. 32, 473–486 (2013).

  33. 33.

    , & Recruitment of the complete hTREX complex is required for Kaposi's sarcoma-associated herpesvirus intronless mRNA nuclear export and virus replication. PLoS Pathogens 4, e1000194 (2008).

  34. 34.

    , , , & Crystal structure of the human ATP-dependent splicing and export factor UAP56. Proc. Natl Acad. Sci. USA 101, 17628–17633 (2004).

  35. 35.

    et al. Novel, potent small-molecule inhibitors of the molecular chaperone Hsp90 discovered through structure-based design. J. Med. Chem. 48, 4212–4215 (2005).

  36. 36.

    , & Biochemical characterization of the ATPase and helicase activity of UAP56, an essential pre-mRNA splicing and mRNA export factor. J. Biol. Chem. 282, 22544–22550 (2007).

  37. 37.

    et al. In vitro biological characterization of a novel, synthetic diaryl pyrazole resorcinol class of heat shock protein 90 inhibitors. Cancer Res. 67, 2206–2216 (2007).

  38. 38.

    & Nucleolar disruption impairs Kaposi's sarcoma-associated herpesvirus ORF57-mediated nuclear export of intronless viral mRNAs. FEBS Lett. 583, 3549–3556 (2009).

  39. 39.

    , & Interactions between KSHV ORF57 and the novel human TREX proteins, CHTOP and CIP29. J. Gen. Virol. 97, 1904–1910 (2016).

  40. 40.

    , , & Viral factors reveal a role for REF/Aly in nuclear RNA stability. Mol. Cell. Biol. 32, 1260–1270 (2012).

  41. 41.

    et al. Global changes in Kaposi's sarcoma-associated virus gene expression patterns following expression of a tetracycline-inducible Rta transactivator. J. Virol. 77, 4205–4220 (2003).

  42. 42.

    , , , & Hsp90 inhibitors are efficacious against Kaposi Sarcoma by enhancing the degradation of the essential viral gene LANA, of the viral co-receptor EphA2 as well as other client proteins. PLoS Pathogens 8, e1003048 (2012).

  43. 43.

    et al. Targeting the Hsp90-associated viral oncoproteome in gammaherpesvirus-associated malignancies. Blood 122, 2837–2847 (2013).

  44. 44.

    et al. The effects of heat shock protein 90 inhibitors on apoptosis and viral replication in primary effusion lymphoma cells. Biol. Pharm. Bull. 35, 725–730 (2012).

  45. 45.

    , , , & Crystal structure of UAP56, a DExD/H-box protein involved in pre-mRNA splicing and mRNA export. Structure 12, 1373–1381 (2004).

  46. 46.

    et al. Mechanism of ATP turnover inhibition in the EJC. RNA 15, 67–75 (2009).

  47. 47.

    et al. Preclinical pharmacokinetics and metabolism of a novel diaryl pyrazole resorcinol series of heat shock protein 90 inhibitors. Mol. Cancer Ther. 5, 1628–1637 (2006).

  48. 48.

    et al. Structural basis for the recognition of cellular mRNA export factor REF by herpes viral proteins HSV-1 ICP27 and HVS ORF57. PLoS Pathogens 7, e1001244 (2011).

  49. 49.

    , , , & The UL69 transactivator protein of human cytomegalovirus interacts with DEXD/H-box RNA helicase UAP56 to promote cytoplasmic accumulation of unspliced RNA. Mol. Cell. Biol. 26, 1631–1643 (2006).

  50. 50.

    & Individual influenza A virus mRNAs show differential dependence on cellular NXF1/TAP for their nuclear export. J. Gen. Virol. 91, 1290–1301 (2010).

  51. 51.

    , , , & ZINC: a free tool to discover chemistry for biology. J. Chem. Inf. Model. 52, 1757–1768 (2012).

  52. 52.

    et al. Human mRNA export machinery recruited to the 5’ end of mRNA. Cell 127, 1389–1400 (2006).

  53. 53.

    , , , & NEDDylation is essential for Kaposi's sarcoma-associated herpesvirus latency and lytic reactivation and represents a novel anti-KSHV target. PLoS Pathogens 11, e1004771 (2015).

  54. 54.

    et al. Merkel cell polyomavirus small T antigen targets the NEMO adaptor protein to disrupt inflammatory signaling. J. Virol. 87, 13853–13867 (2013).

  55. 55.

    et al. Merkel cell polyomavirus small T antigen mediates microtubule destabilisation to promote cell motility and migration. J. Virol. 89, 35–47 (2015).

Download references

Acknowledgements

The authors thank J. Jung (UCLA), S. Efstathiou and J. Sinclair (Cambridge) for cell lines and recombinant viruses, and S. Wilson (Sheffield), R. Reed (Harvard), T. Stamminger (Erlangen) and R.M. Sandri-Goldin (Irvine) for the gift of antibodies and plasmid constructs. The authors also thank I. Mainfield, Centre for Biomolecular Interactions, Faculty of Biological Sciences, for advice. This work was supported in part by the Wellcome Trust (093788/Z/10/Z); Worldwide Cancer Research (12-1045), BBSRC (BB/000306; BB/M006557) and CRUK (C12057/A19430).

Author information

Affiliations

  1. School of Molecular and Cellular Biology, Leeds LS2 9JT, UK

    • Sophie Schumann
    • , Brian R. Jackson
    •  & Adrian Whitehouse
  2. Astbury Centre for Structural Molecular Biology, Leeds LS2 9JT, UK

    • Sophie Schumann
    • , Brian R. Jackson
    • , Richard Foster
    •  & Adrian Whitehouse
  3. School of Chemistry, University of Leeds, Leeds LS2 9JT, UK

    • Ian Yule
    • , Steven K. Whitehead
    • , Charlotte Revill
    •  & Richard Foster

Authors

  1. Search for Sophie Schumann in:

  2. Search for Brian R. Jackson in:

  3. Search for Ian Yule in:

  4. Search for Steven K. Whitehead in:

  5. Search for Charlotte Revill in:

  6. Search for Richard Foster in:

  7. Search for Adrian Whitehouse in:

Contributions

S.S. designed and performed experiments, analysed data and wrote the manuscript. B.R.J. performed experiments and analysed data. I.Y. performed the virtual high-throughput screening and modelling. S.K.W. and C.R. synthesized tested compounds. R.F. and A.W. designed and performed experiments, analysed data and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Richard Foster or Adrian Whitehouse.

Supplementary information

PDF files

  1. 1.

    Supplementary information

    Supplementary Figures 1–14