skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Middle East Respiratory Syndrome Coronavirus Nonstructural Protein 16 Is Necessary for Interferon Resistance and Viral Pathogenesis

Abstract

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'O-methyltransferase (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'O-MTase 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 andin 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 conservedmore » universal target for rapid live attenuated vaccine design in an expanding CoV outbreak setting. IMPORTANCECoronavirus (CoV) emergence in both humans and livestock represents a significant threat to global public health, as evidenced by the sudden emergence of severe acute respiratory syndrome CoV (SARS-CoV), MERS-CoV, porcine epidemic diarrhea virus, and swine delta CoV in the 21st century. These studies describe an approach that effectively targets the highly conserved 2'O-MTase activity of CoVs for attenuation. With clear understanding of the IFN/IFIT (IFN-induced proteins with tetratricopeptide repeats)-based mechanism, NSP16 mutants provide a suitable target for a live attenuated vaccine platform, as well as therapeutic development for both current and future emergent CoV strains. Importantly, other approaches targeting other conserved pan-CoV functions have not yet proven effective against MERS-CoV, illustrating the broad applicability of targeting viral 2'O-MTase function across CoVs.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1414531
Report Number(s):
PNNL-SA-128149
Journal ID: ISSN 2379-5042; WN9030198
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Journal Name:
mSphere
Additional Journal Information:
Journal Volume: 2; Journal Issue: 6; Journal ID: ISSN 2379-5042
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; MERS-CoV; NSP16; IFN; viral; pathogenesis

Citation Formats

Menachery, Vineet D., 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., and Fernandez-Sesma, Ana. Middle East Respiratory Syndrome Coronavirus Nonstructural Protein 16 Is Necessary for Interferon Resistance and Viral Pathogenesis. United States: N. p., 2017. Web. doi:10.1128/mSphere.00346-17.
Menachery, Vineet D., 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., & Fernandez-Sesma, Ana. Middle East Respiratory Syndrome Coronavirus Nonstructural Protein 16 Is Necessary for Interferon Resistance and Viral Pathogenesis. United States. doi:10.1128/mSphere.00346-17.
Menachery, Vineet D., 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., and Fernandez-Sesma, Ana. Wed . "Middle East Respiratory Syndrome Coronavirus Nonstructural Protein 16 Is Necessary for Interferon Resistance and Viral Pathogenesis". United States. doi:10.1128/mSphere.00346-17.
@article{osti_1414531,
title = {Middle East Respiratory Syndrome Coronavirus Nonstructural Protein 16 Is Necessary for Interferon Resistance and Viral Pathogenesis},
author = {Menachery, Vineet D. and Gralinski, Lisa E. and Mitchell, Hugh D. and Dinnon, Kenneth H. and Leist, Sarah R. and Yount, Boyd L. and Graham, Rachel L. and McAnarney, Eileen T. and Stratton, Kelly G. and Cockrell, Adam S. and Debbink, Kari and Sims, Amy C. and Waters, Katrina M. and Baric, Ralph S. and Fernandez-Sesma, Ana},
abstractNote = {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'O-methyltransferase (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'O-MTase 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 andin 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. IMPORTANCECoronavirus (CoV) emergence in both humans and livestock represents a significant threat to global public health, as evidenced by the sudden emergence of severe acute respiratory syndrome CoV (SARS-CoV), MERS-CoV, porcine epidemic diarrhea virus, and swine delta CoV in the 21st century. These studies describe an approach that effectively targets the highly conserved 2'O-MTase activity of CoVs for attenuation. With clear understanding of the IFN/IFIT (IFN-induced proteins with tetratricopeptide repeats)-based mechanism, NSP16 mutants provide a suitable target for a live attenuated vaccine platform, as well as therapeutic development for both current and future emergent CoV strains. Importantly, other approaches targeting other conserved pan-CoV functions have not yet proven effective against MERS-CoV, illustrating the broad applicability of targeting viral 2'O-MTase function across CoVs.},
doi = {10.1128/mSphere.00346-17},
journal = {mSphere},
issn = {2379-5042},
number = 6,
volume = 2,
place = {United States},
year = {2017},
month = {11}
}

Works referenced in this record:

Middle East respiratory syndrome coronavirus vaccines: current status and novel approaches
journal, April 2017


Infidelity of SARS-CoV Nsp14-Exonuclease Mutant Virus Replication Is Revealed by Complete Genome Sequencing
journal, May 2010


SARS coronavirus pathogenesis: host innate immune responses and viral antagonism of interferon
journal, June 2012


Reverse genetics with a full-length infectious cDNA of the Middle East respiratory syndrome coronavirus
journal, September 2013

  • Scobey, T.; Yount, B. L.; Sims, A. C.
  • Proceedings of the National Academy of Sciences, Vol. 110, Issue 40
  • DOI: 10.1073/pnas.1311542110

Coronavirus nonstructural protein 15 mediates evasion of dsRNA sensors and limits apoptosis in macrophages
journal, May 2017

  • Deng, Xufang; Hackbart, Matthew; Mettelman, Robert C.
  • Proceedings of the National Academy of Sciences, Vol. 114, Issue 21
  • DOI: 10.1073/pnas.1618310114

Middle East respiratory syndrome
journal, September 2015


Polymerases of hepatitis C viruses and flaviviruses: Structural and mechanistic insights and drug development
journal, May 2014


Ribose 2′-O-methylation provides a molecular signature for the distinction of self and non-self mRNA dependent on the RNA sensor Mda5
journal, January 2011

  • Züst, Roland; Cervantes-Barragan, Luisa; Habjan, Matthias
  • Nature Immunology, Vol. 12, Issue 2
  • DOI: 10.1038/ni.1979

A mouse model for MERS coronavirus-induced acute respiratory distress syndrome
journal, November 2016


SARS-like WIV1-CoV poised for human emergence
journal, March 2016

  • Menachery, Vineet D.; Yount, Boyd L.; Sims, Amy C.
  • Proceedings of the National Academy of Sciences, Vol. 113, Issue 11
  • DOI: 10.1073/pnas.1517719113

Host Regulatory Network Response to Infection with Highly Pathogenic H5N1 Avian Influenza Virus
journal, August 2011

  • Li, C.; Bankhead, A.; Eisfeld, A. J.
  • Journal of Virology, Vol. 85, Issue 21
  • DOI: 10.1128/JVI.05792-11

Release of Severe Acute Respiratory Syndrome Coronavirus Nuclear Import Block Enhances Host Transcription in Human Lung Cells
journal, January 2013

  • Sims, A. C.; Tilton, S. C.; Menachery, V. D.
  • Journal of Virology, Vol. 87, Issue 7
  • DOI: 10.1128/JVI.02520-12

Jumping species—a mechanism for coronavirus persistence and survival
journal, April 2017


Coronavirus non-structural protein 16: Evasion, attenuation, and possible treatments
journal, December 2014


Identification of the Mechanisms Causing Reversion to Virulence in an Attenuated SARS-CoV for the Design of a Genetically Stable Vaccine
journal, October 2015

  • Jimenez-Guardeño, Jose M.; Regla-Nava, Jose A.; Nieto-Torres, Jose L.
  • PLOS Pathogens, Vol. 11, Issue 10
  • DOI: 10.1371/journal.ppat.1005215

Attenuation and Restoration of Severe Acute Respiratory Syndrome Coronavirus Mutant Lacking 2'-O-Methyltransferase Activity
journal, January 2014

  • Menachery, V. D.; Yount, B. L.; Josset, L.
  • Journal of Virology, Vol. 88, Issue 8
  • DOI: 10.1128/JVI.03571-13

Prophylactic and postexposure efficacy of a potent human monoclonal antibody against MERS coronavirus
journal, July 2015

  • Corti, Davide; Zhao, Jincun; Pedotti, Mattia
  • Proceedings of the National Academy of Sciences, Vol. 112, Issue 33
  • DOI: 10.1073/pnas.1510199112

A live, impaired-fidelity coronavirus vaccine protects in an aged, immunocompromised mouse model of lethal disease
journal, November 2012

  • Graham, Rachel L.; Becker, Michelle M.; Eckerle, Lance D.
  • Nature Medicine, Vol. 18, Issue 12
  • DOI: 10.1038/nm.2972

A SARS-like cluster of circulating bat coronaviruses shows potential for human emergence
journal, November 2015

  • Menachery, Vineet D.; Yount, Boyd L.; Debbink, Kari
  • Nature Medicine, Vol. 21, Issue 12
  • DOI: 10.1038/nm.3985

Engineering a Replication-Competent, Propagation-Defective Middle East Respiratory Syndrome Coronavirus as a Vaccine Candidate
journal, September 2013