Rescue of infectious Arumowot virus from cloned cDNA: Posttranslational degradation of Arumowot virus NSs protein in human cells
Identifieur interne : 000421 ( Pmc/Checkpoint ); précédent : 000420; suivant : 000422Rescue of infectious Arumowot virus from cloned cDNA: Posttranslational degradation of Arumowot virus NSs protein in human cells
Auteurs : Hoai J. Hallam [États-Unis] ; Nandadeva Lokugamage [États-Unis] ; Tetsuro Ikegami [États-Unis]Source :
- PLoS Neglected Tropical Diseases [ 1935-2727 ] ; 2019.
Abstract
Rift Valley fever (RVF) is a mosquito-borne zoonotic disease endemic to Africa and the Middle East, affecting both humans and ruminants. There are no licensed vaccines or antivirals available for humans, whereas research using RVF virus (RVFV) is strictly regulated in many countries with safety concerns. Nonpathogenic Arumowot virus (AMTV), a mosquito-borne phlebovirus in Africa, is likely useful for the screening of broad-acting antiviral candidates for phleboviruses including RVFV, as well as a potential vaccine vector for RVF. In this study, we aimed to generate T7 RNA polymerase-driven reverse genetics system for AMTV. We hypothesized that recombinant AMTV (rAMTV) is viable, and AMTV NSs protein is dispensable for efficient replication of rAMTV in type-I interferon (IFN)-incompetent cells, whereas AMTV NSs proteins support robust viral replication in type-I IFN-competent cells. The study demonstrated the rescue of rAMTV and that lacking the NSs gene (rAMTVΔNSs), that expressing green fluorescent protein (GFP) (rAMTV-GFP) or that expressing
Url:
DOI: 10.1371/journal.pntd.0007904
PubMed: 31751340
PubMed Central: 6894884
Affiliations:
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Hallam</name><affiliation wicri:level="2"><nlm:aff id="aff001"><addr-line>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Lokugamage, Nandadeva" sort="Lokugamage, Nandadeva" uniqKey="Lokugamage N" first="Nandadeva" last="Lokugamage">Nandadeva Lokugamage</name><affiliation wicri:level="2"><nlm:aff id="aff001"><addr-line>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Ikegami, Tetsuro" sort="Ikegami, Tetsuro" uniqKey="Ikegami T" first="Tetsuro" last="Ikegami">Tetsuro Ikegami</name><affiliation wicri:level="2"><nlm:aff id="aff001"><addr-line>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation><affiliation wicri:level="2"><nlm:aff id="aff002"><addr-line>Sealy Institute for Vaccine Sciences, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Sealy Institute for Vaccine Sciences, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation><affiliation wicri:level="2"><nlm:aff id="aff003"><addr-line>Center for Biodefense and Emerging Infectious Diseases, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Center for Biodefense and Emerging Infectious Diseases, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">31751340</idno><idno type="pmc">6894884</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6894884</idno><idno type="RBID">PMC:6894884</idno><idno type="doi">10.1371/journal.pntd.0007904</idno><date when="2019">2019</date><idno type="wicri:Area/Pmc/Corpus">000769</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000769</idno><idno type="wicri:Area/Pmc/Curation">000769</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000769</idno><idno type="wicri:Area/Pmc/Checkpoint">000421</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000421</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Rescue of infectious Arumowot virus from cloned cDNA: Posttranslational degradation of Arumowot virus NSs protein in human cells</title><author><name sortKey="Hallam, Hoai J" sort="Hallam, Hoai J" uniqKey="Hallam H" first="Hoai J." last="Hallam">Hoai J. Hallam</name><affiliation wicri:level="2"><nlm:aff id="aff001"><addr-line>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Lokugamage, Nandadeva" sort="Lokugamage, Nandadeva" uniqKey="Lokugamage N" first="Nandadeva" last="Lokugamage">Nandadeva Lokugamage</name><affiliation wicri:level="2"><nlm:aff id="aff001"><addr-line>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author><author><name sortKey="Ikegami, Tetsuro" sort="Ikegami, Tetsuro" uniqKey="Ikegami T" first="Tetsuro" last="Ikegami">Tetsuro Ikegami</name><affiliation wicri:level="2"><nlm:aff id="aff001"><addr-line>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation><affiliation wicri:level="2"><nlm:aff id="aff002"><addr-line>Sealy Institute for Vaccine Sciences, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Sealy Institute for Vaccine Sciences, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation><affiliation wicri:level="2"><nlm:aff id="aff003"><addr-line>Center for Biodefense and Emerging Infectious Diseases, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Center for Biodefense and Emerging Infectious Diseases, The University of Texas Medical Branch at Galveston, Galveston, Texas</wicri:regionArea><placeName><region type="state">Texas</region></placeName></affiliation></author></analytic><series><title level="j">PLoS Neglected Tropical Diseases</title><idno type="ISSN">1935-2727</idno><idno type="eISSN">1935-2735</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Rift Valley fever (RVF) is a mosquito-borne zoonotic disease endemic to Africa and the Middle East, affecting both humans and ruminants. There are no licensed vaccines or antivirals available for humans, whereas research using RVF virus (RVFV) is strictly regulated in many countries with safety concerns. Nonpathogenic Arumowot virus (AMTV), a mosquito-borne phlebovirus in Africa, is likely useful for the screening of broad-acting antiviral candidates for phleboviruses including RVFV, as well as a potential vaccine vector for RVF. In this study, we aimed to generate T7 RNA polymerase-driven reverse genetics system for AMTV. We hypothesized that recombinant AMTV (rAMTV) is viable, and AMTV NSs protein is dispensable for efficient replication of rAMTV in type-I interferon (IFN)-incompetent cells, whereas AMTV NSs proteins support robust viral replication in type-I IFN-competent cells. The study demonstrated the rescue of rAMTV and that lacking the NSs gene (rAMTVΔNSs), that expressing green fluorescent protein (GFP) (rAMTV-GFP) or that expressing <italic>Renilla</italic> luciferase (rAMTV-rLuc) from cloned cDNA. The rAMTV-rLuc and the RVFV rMP12-rLuc showed a similar susceptibility to favipiravir or ribavirin. Interestingly, neither of rAMTV nor rAMTVΔNSs replicated efficiently in human MRC-5 or A549 cells, regardless of the presence of NSs gene. Little accumulation of AMTV NSs protein occurred in those cells, which was restored via treatment with proteasomal inhibitor MG132. In murine MEF or Hepa1-6 cells, rAMTV, but not rAMTVΔNSs, replicated efficiently, with an inhibition of IFN-β gene upregulation. This study showed an establishment of the first reverse genetics for AMTV, a lack of stability of AMTV NSs proteins in human cells, and an IFN-β gene antagonist function of AMTV NSs proteins in murine cells. 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article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">PLoS Negl Trop Dis</journal-id><journal-id journal-id-type="iso-abbrev">PLoS Negl Trop Dis</journal-id><journal-id journal-id-type="publisher-id">plos</journal-id><journal-id journal-id-type="pmc">plosntds</journal-id><journal-title-group><journal-title>PLoS Neglected Tropical Diseases</journal-title></journal-title-group><issn pub-type="ppub">1935-2727</issn><issn pub-type="epub">1935-2735</issn><publisher><publisher-name>Public Library of Science</publisher-name><publisher-loc>San Francisco, CA USA</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">31751340</article-id><article-id pub-id-type="pmc">6894884</article-id><article-id pub-id-type="doi">10.1371/journal.pntd.0007904</article-id><article-id pub-id-type="publisher-id">PNTD-D-19-00748</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and life sciences</subject><subj-group><subject>Organisms</subject><subj-group><subject>Viruses</subject><subj-group><subject>RNA viruses</subject><subj-group><subject>Bunyaviruses</subject><subj-group><subject>Rift Valley fever virus</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and life sciences</subject><subj-group><subject>Microbiology</subject><subj-group><subject>Medical microbiology</subject><subj-group><subject>Microbial pathogens</subject><subj-group><subject>Viral pathogens</subject><subj-group><subject>Bunyaviruses</subject><subj-group><subject>Rift Valley fever virus</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and health sciences</subject><subj-group><subject>Pathology and laboratory medicine</subject><subj-group><subject>Pathogens</subject><subj-group><subject>Microbial pathogens</subject><subj-group><subject>Viral pathogens</subject><subj-group><subject>Bunyaviruses</subject><subj-group><subject>Rift Valley fever virus</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and life sciences</subject><subj-group><subject>Organisms</subject><subj-group><subject>Viruses</subject><subj-group><subject>Viral pathogens</subject><subj-group><subject>Bunyaviruses</subject><subj-group><subject>Rift Valley fever virus</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Research and Analysis Methods</subject><subj-group><subject>Biological Cultures</subject><subj-group><subject>Cell Lines</subject><subj-group><subject>Vero Cells</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and Health Sciences</subject><subj-group><subject>Pharmacology</subject><subj-group><subject>Drugs</subject><subj-group><subject>Antimicrobials</subject><subj-group><subject>Antivirals</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Microbiology</subject><subj-group><subject>Microbial Control</subject><subj-group><subject>Antimicrobials</subject><subj-group><subject>Antivirals</subject></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Microbiology</subject><subj-group><subject>Virology</subject><subj-group><subject>Antivirals</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Microbiology</subject><subj-group><subject>Virology</subject><subj-group><subject>Viral Replication</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Research and analysis methods</subject><subj-group><subject>Extraction techniques</subject><subj-group><subject>RNA extraction</subject></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and Life Sciences</subject><subj-group><subject>Molecular Biology</subject><subj-group><subject>Molecular Biology Techniques</subject><subj-group><subject>Reverse Genetics</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Research and Analysis Methods</subject><subj-group><subject>Molecular Biology Techniques</subject><subj-group><subject>Reverse Genetics</subject></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and health sciences</subject><subj-group><subject>Tropical diseases</subject><subj-group><subject>Neglected tropical diseases</subject><subj-group><subject>Rift Valley fever</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and health sciences</subject><subj-group><subject>Infectious diseases</subject><subj-group><subject>Viral diseases</subject><subj-group><subject>Rift Valley fever</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Medicine and health sciences</subject><subj-group><subject>Infectious diseases</subject><subj-group><subject>Zoonoses</subject><subj-group><subject>Rift Valley fever</subject></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Biology and life sciences</subject><subj-group><subject>Molecular biology</subject><subj-group><subject>Molecular biology techniques</subject><subj-group><subject>Cloning</subject><subj-group><subject>DNA cloning</subject><subj-group><subject>Complementary DNA cloning</subject></subj-group></subj-group></subj-group></subj-group></subj-group></subj-group><subj-group subj-group-type="Discipline-v3"><subject>Research and analysis methods</subject><subj-group><subject>Molecular biology techniques</subject><subj-group><subject>Cloning</subject><subj-group><subject>DNA cloning</subject><subj-group><subject>Complementary DNA cloning</subject></subj-group></subj-group></subj-group></subj-group></subj-group></article-categories><title-group><article-title>Rescue of infectious Arumowot virus from cloned cDNA: Posttranslational degradation of Arumowot virus NSs protein in human cells</article-title><alt-title alt-title-type="running-head">Reverse genetics for Arumowot virus</alt-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Hallam</surname><given-names>Hoai J.</given-names></name><role content-type="http://credit.casrai.org/">Funding acquisition</role><role content-type="http://credit.casrai.org/">Investigation</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Validation</role><role content-type="http://credit.casrai.org/">Writing – original draft</role><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref ref-type="author-notes" rid="currentaff001"><sup>¤a</sup></xref></contrib><contrib contrib-type="author"><name><surname>Lokugamage</surname><given-names>Nandadeva</given-names></name><role content-type="http://credit.casrai.org/">Investigation</role><role content-type="http://credit.casrai.org/">Methodology</role><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref ref-type="author-notes" rid="currentaff002"><sup>¤b</sup></xref></contrib><contrib contrib-type="author"><contrib-id authenticated="true" contrib-id-type="orcid">http://orcid.org/0000-0001-8318-2783</contrib-id><name><surname>Ikegami</surname><given-names>Tetsuro</given-names></name><role content-type="http://credit.casrai.org/">Conceptualization</role><role content-type="http://credit.casrai.org/">Funding acquisition</role><role content-type="http://credit.casrai.org/">Investigation</role><role content-type="http://credit.casrai.org/">Methodology</role><role content-type="http://credit.casrai.org/">Project administration</role><role content-type="http://credit.casrai.org/">Resources</role><role content-type="http://credit.casrai.org/">Supervision</role><role content-type="http://credit.casrai.org/">Validation</role><role content-type="http://credit.casrai.org/">Writing – original draft</role><role content-type="http://credit.casrai.org/">Writing – review & editing</role><xref ref-type="aff" rid="aff001"><sup>1</sup></xref><xref ref-type="aff" rid="aff002"><sup>2</sup></xref><xref ref-type="aff" rid="aff003"><sup>3</sup></xref><xref ref-type="corresp" rid="cor001">*</xref></contrib></contrib-group><aff id="aff001"><label>1</label><addr-line>Department of Pathology, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></aff><aff id="aff002"><label>2</label><addr-line>Sealy Institute for Vaccine Sciences, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></aff><aff id="aff003"><label>3</label><addr-line>Center for Biodefense and Emerging Infectious Diseases, The University of Texas Medical Branch at Galveston, Galveston, Texas, United States of America</addr-line></aff><contrib-group><contrib contrib-type="editor"><name><surname>Samy</surname><given-names>Abdallah M.</given-names></name><role>Editor</role><xref ref-type="aff" rid="edit1"></xref></contrib></contrib-group><aff id="edit1"><addr-line>Faculty of Science, Ain Shams University (ASU), EGYPT</addr-line></aff><author-notes><fn fn-type="COI-statement" id="coi001"><p>The authors have declared that no competing interests exist.</p></fn><fn fn-type="current-aff" id="currentaff001"><label>¤a</label><p>Current address: Department of Oncology, The University of Texas at Austin Dell Medical School, Austin, TX, United States of America</p></fn><fn fn-type="current-aff" id="currentaff002"><label>¤b</label><p>Current address: Department of Microbiology and Immunology, The University of Texas Medical Branch at Galveston, TX, United States of America</p></fn><corresp id="cor001">* E-mail: <email>teikegam@utmb.edu</email></corresp></author-notes><pub-date pub-type="epub"><day>21</day><month>11</month><year>2019</year></pub-date><pub-date pub-type="collection"><month>11</month><year>2019</year></pub-date><volume>13</volume><issue>11</issue><elocation-id>e0007904</elocation-id><history><date date-type="received"><day>6</day><month>5</month><year>2019</year></date><date date-type="accepted"><day>4</day><month>11</month><year>2019</year></date></history><permissions><copyright-statement>© 2019 Hallam et al</copyright-statement><copyright-year>2019</copyright-year><copyright-holder>Hallam et al</copyright-holder><license xlink:href="http://creativecommons.org/licenses/by/4.0/"><license-p>This is an open access article distributed under the terms of the <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">Creative Commons Attribution License</ext-link>, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.</license-p></license></permissions><self-uri content-type="pdf" xlink:href="pntd.0007904.pdf"></self-uri><abstract><p>Rift Valley fever (RVF) is a mosquito-borne zoonotic disease endemic to Africa and the Middle East, affecting both humans and ruminants. There are no licensed vaccines or antivirals available for humans, whereas research using RVF virus (RVFV) is strictly regulated in many countries with safety concerns. Nonpathogenic Arumowot virus (AMTV), a mosquito-borne phlebovirus in Africa, is likely useful for the screening of broad-acting antiviral candidates for phleboviruses including RVFV, as well as a potential vaccine vector for RVF. In this study, we aimed to generate T7 RNA polymerase-driven reverse genetics system for AMTV. We hypothesized that recombinant AMTV (rAMTV) is viable, and AMTV NSs protein is dispensable for efficient replication of rAMTV in type-I interferon (IFN)-incompetent cells, whereas AMTV NSs proteins support robust viral replication in type-I IFN-competent cells. The study demonstrated the rescue of rAMTV and that lacking the NSs gene (rAMTVΔNSs), that expressing green fluorescent protein (GFP) (rAMTV-GFP) or that expressing <italic>Renilla</italic> luciferase (rAMTV-rLuc) from cloned cDNA. The rAMTV-rLuc and the RVFV rMP12-rLuc showed a similar susceptibility to favipiravir or ribavirin. Interestingly, neither of rAMTV nor rAMTVΔNSs replicated efficiently in human MRC-5 or A549 cells, regardless of the presence of NSs gene. Little accumulation of AMTV NSs protein occurred in those cells, which was restored via treatment with proteasomal inhibitor MG132. In murine MEF or Hepa1-6 cells, rAMTV, but not rAMTVΔNSs, replicated efficiently, with an inhibition of IFN-β gene upregulation. This study showed an establishment of the first reverse genetics for AMTV, a lack of stability of AMTV NSs proteins in human cells, and an IFN-β gene antagonist function of AMTV NSs proteins in murine cells. The AMTV can be a nonpathogenic surrogate model for studying phleboviruses including RVFV.</p></abstract><abstract abstract-type="summary"><title>Author summary</title><p>Rift Valley fever virus (RVFV) is a mosquito-borne phlebovirus endemic to Africa and the Middle East, causing devastating outbreaks affecting both humans and animals. The reverse genetics system for RVFV has contributed to the virology, vaccinology, and antiviral screening for RVFV. In this study, we generated the first reverse genetics system for a mosquito-borne nonpathogenic phlebovirus (Arumowot virus; AMTV) endemic to Africa, which is phylogenetically related to RVFV. The generation of recombinant AMTV supports the screening of broad-acting antivirals and vaccine development for RVF. The nonstructural NSs protein is known as a major virulence factor for RVF, yet this study revealed that AMTV NSs protein was rapidly degraded in human cells via cellular proteasomes. In contrast, AMTV NSs protein functioned as an antagonist of interferon-β gene upregulation in murine cells. The AMTV can be a nonpathogenic surrogate model for studying phleboviruses including RVFV.</p></abstract><funding-group><award-group id="award001"><funding-source><institution>the Institute for Human Infections and Immunity</institution></funding-source><principal-award-recipient><contrib-id authenticated="true" contrib-id-type="orcid">http://orcid.org/0000-0001-8318-2783</contrib-id><name><surname>Ikegami</surname><given-names>Tetsuro</given-names></name></principal-award-recipient></award-group><award-group id="award002"><funding-source><institution>the United States Agency for International development</institution></funding-source><award-id>subcontract 26-3002-6062</award-id><principal-award-recipient><contrib-id authenticated="true" contrib-id-type="orcid">http://orcid.org/0000-0001-8318-2783</contrib-id><name><surname>Ikegami</surname><given-names>Tetsuro</given-names></name></principal-award-recipient></award-group><award-group id="award003"><funding-source><institution>the Sealy Institute for Vaccine Sciences</institution></funding-source><principal-award-recipient><name><surname>Hallam</surname><given-names>Hoai J.</given-names></name></principal-award-recipient></award-group><award-group id="award004"><funding-source><institution>the Jeane B. Kempner Scholar Award Fund</institution></funding-source><principal-award-recipient><name><surname>Hallam</surname><given-names>Hoai J.</given-names></name></principal-award-recipient></award-group><funding-statement>TI was supported by Data Collection grant from the Institute for Human Infections and Immunity (<ext-link ext-link-type="uri" xlink:href="https://www.utmb.edu/ihii">https://www.utmb.edu/ihii</ext-link>) at UTMB, and partly supported by the subcontract 26-3002-6062 from the University of Texas El Paso, through the United States Agency for International development (USAID, AID-OAA-A-13-00084: This contents do not necessarily reflect the views of USAID or the U.S. government). HJH was supported by the Sealy Institute for Vaccine Sciences (<ext-link ext-link-type="uri" xlink:href="https://www.utmb.edu/sivs">https://www.utmb.edu/sivs</ext-link>) Predoctoral fellowship, as well as the Jeane B. Kempner Predoctoral Fellowship (<ext-link ext-link-type="uri" xlink:href="https://gsbs.utmb.edu/kempner-fellowship">https://gsbs.utmb.edu/kempner-fellowship</ext-link>) at UTMB. Funders did not play any role in study design, data collection, analysis, decision to publish, or preparation of the manuscript.</funding-statement></funding-group><counts><fig-count count="7"></fig-count><table-count count="0"></table-count><page-count count="21"></page-count></counts><custom-meta-group><custom-meta><meta-name>PLOS Publication Stage</meta-name><meta-value>vor-update-to-uncorrected-proof</meta-value></custom-meta><custom-meta><meta-name>Publication Update</meta-name><meta-value>2019-12-05</meta-value></custom-meta><custom-meta id="data-availability"><meta-name>Data Availability</meta-name><meta-value>All relevant data are within the manuscript.</meta-value></custom-meta></custom-meta-group></article-meta><notes><title>Data Availability</title><p>All relevant data are within the manuscript.</p></notes></front></pmc><affiliations><list><country><li>États-Unis</li></country><region><li>Texas</li></region></list><tree><country name="États-Unis"><region name="Texas"><name sortKey="Hallam, Hoai J" sort="Hallam, Hoai J" uniqKey="Hallam H" first="Hoai J." last="Hallam">Hoai J. Hallam</name></region><name sortKey="Ikegami, Tetsuro" sort="Ikegami, Tetsuro" uniqKey="Ikegami T" first="Tetsuro" last="Ikegami">Tetsuro Ikegami</name><name sortKey="Ikegami, Tetsuro" sort="Ikegami, Tetsuro" uniqKey="Ikegami T" first="Tetsuro" last="Ikegami">Tetsuro Ikegami</name><name sortKey="Ikegami, Tetsuro" sort="Ikegami, Tetsuro" uniqKey="Ikegami T" first="Tetsuro" last="Ikegami">Tetsuro Ikegami</name><name sortKey="Lokugamage, Nandadeva" sort="Lokugamage, Nandadeva" uniqKey="Lokugamage N" first="Nandadeva" last="Lokugamage">Nandadeva Lokugamage</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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