Middle East respiratory syndrome coronavirus nonstructural protein 16 is necessary for interferon resistance and viral pathogenesis

Vineet Menachery, Lisa E. Gralinski, Hugh D. Mitchell, Kenneth H. Dinnon, Sarah R. Leist, Boyd L. Yount, Rachel L. Graham, Eileen T. McAnarney, Kelly G. Stratton, Adam S. Cockrell, Kari Debbink, Amy C. Sims, Katrina M. Waters, Ralph S. Baric

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Coronaviruses (CoVs) encode a mixture of highly conserved and novel genes, as well as genetic elements necessary for infection and pathogenesis, raising the possibility of common targets for attenuation and therapeutic design. In this study, we focused on highly conserved nonstructural protein 16 (NSP16), a viral 2'Omethyltransferase (2'O-MTase) that encodes critical functions in immune modulation and infection. Using reverse genetics, we disrupted a key motif in the conserved KDKE motif of Middle East respiratory syndrome CoV (MERS-CoV) NSP16 (D130A) and evaluated the effect on viral infection and pathogenesis. While the absence of 2'OMTase activity had only a marginal impact on propagation and replication in Vero cells, dNSP16 mutant MERS-CoV demonstrated significant attenuation relative to the control both in primary human airway cell cultures and in vivo. Further examination indicated that dNSP16 mutant MERS-CoV had a type I interferon (IFN)-based attenuation and was partially restored in the absence of molecules of IFN-induced proteins with tetratricopeptide repeats. Importantly, the robust attenuation permitted the use of dNSP16 mutant MERS-CoV as a live attenuated vaccine platform protecting from a challenge with a mouse-adapted MERS-CoV strain. These studies demonstrate the importance of the conserved 2'O-MTase activity for CoV pathogenesis and highlight NSP16 as a conserved universal target for rapid live attenuated vaccine design in an expanding CoV outbreak setting.

Original languageEnglish (US)
Article numbere00346-17
JournalmSphere
Volume2
Issue number6
DOIs
StatePublished - Nov 1 2017

Fingerprint

Coronavirus Infections
Interferons
Attenuated Vaccines
Proteins
Reverse Genetics
Interferon Type I
Coronavirus
Vero Cells
Virus Diseases
Infection
Disease Outbreaks
Cell Culture Techniques
coronavirus nonstructural protein
Middle East Respiratory Syndrome Coronavirus
Genes

Keywords

  • Coronavirus
  • Emerging virus
  • IFIT
  • Live attenuated
  • MERS-CoV
  • Methyltransferase
  • SARS-CoV
  • Vaccine

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Menachery, V., Gralinski, L. E., Mitchell, H. D., Dinnon, K. H., Leist, S. R., Yount, B. L., ... Baric, R. S. (2017). Middle East respiratory syndrome coronavirus nonstructural protein 16 is necessary for interferon resistance and viral pathogenesis. mSphere, 2(6), [e00346-17]. https://doi.org/10.1128/mSphere.00346-17

Middle East respiratory syndrome coronavirus nonstructural protein 16 is necessary for interferon resistance and viral pathogenesis. / Menachery, Vineet; Gralinski, Lisa E.; Mitchell, Hugh D.; Dinnon, Kenneth H.; Leist, Sarah R.; Yount, Boyd L.; Graham, Rachel L.; McAnarney, Eileen T.; Stratton, Kelly G.; Cockrell, Adam S.; Debbink, Kari; Sims, Amy C.; Waters, Katrina M.; Baric, Ralph S.

In: mSphere, Vol. 2, No. 6, e00346-17, 01.11.2017.

Research output: Contribution to journalArticle

Menachery, V, Gralinski, LE, Mitchell, HD, Dinnon, KH, Leist, SR, Yount, BL, Graham, RL, McAnarney, ET, Stratton, KG, Cockrell, AS, Debbink, K, Sims, AC, Waters, KM & Baric, RS 2017, 'Middle East respiratory syndrome coronavirus nonstructural protein 16 is necessary for interferon resistance and viral pathogenesis', mSphere, vol. 2, no. 6, e00346-17. https://doi.org/10.1128/mSphere.00346-17
Menachery, Vineet ; Gralinski, Lisa E. ; Mitchell, Hugh D. ; Dinnon, Kenneth H. ; Leist, Sarah R. ; Yount, Boyd L. ; Graham, Rachel L. ; McAnarney, Eileen T. ; Stratton, Kelly G. ; Cockrell, Adam S. ; Debbink, Kari ; Sims, Amy C. ; Waters, Katrina M. ; Baric, Ralph S. / Middle East respiratory syndrome coronavirus nonstructural protein 16 is necessary for interferon resistance and viral pathogenesis. In: mSphere. 2017 ; Vol. 2, No. 6.
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abstract = "Coronaviruses (CoVs) encode a mixture of highly conserved and novel genes, as well as genetic elements necessary for infection and pathogenesis, raising the possibility of common targets for attenuation and therapeutic design. In this study, we focused on highly conserved nonstructural protein 16 (NSP16), a viral 2'Omethyltransferase (2'O-MTase) that encodes critical functions in immune modulation and infection. Using reverse genetics, we disrupted a key motif in the conserved KDKE motif of Middle East respiratory syndrome CoV (MERS-CoV) NSP16 (D130A) and evaluated the effect on viral infection and pathogenesis. While the absence of 2'OMTase activity had only a marginal impact on propagation and replication in Vero cells, dNSP16 mutant MERS-CoV demonstrated significant attenuation relative to the control both in primary human airway cell cultures and in vivo. Further examination indicated that dNSP16 mutant MERS-CoV had a type I interferon (IFN)-based attenuation and was partially restored in the absence of molecules of IFN-induced proteins with tetratricopeptide repeats. Importantly, the robust attenuation permitted the use of dNSP16 mutant MERS-CoV as a live attenuated vaccine platform protecting from a challenge with a mouse-adapted MERS-CoV strain. These studies demonstrate the importance of the conserved 2'O-MTase activity for CoV pathogenesis and highlight NSP16 as a conserved universal target for rapid live attenuated vaccine design in an expanding CoV outbreak setting.",
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AU - Cockrell, Adam S.

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