{"id":{"repo_id":"utmb","oai_identifier":"oai:utmb-ir.tdl.org:2152.3/11878"},"canonical_url":"https://search.dev.ndltd.org/etd/utmb/oai:utmb-ir.tdl.org:2152.3/11878","repository":{"repo_id":"utmb","name":"University of Texas Medical Branch","base_url":"https://utmb-ir.tdl.org/server/oai/request"},"display":{"title":"Subversion of innate antiviral immunity by SARS-CoV-2: An example from the study of nonstructural protein 16.","abstract":"Understanding the molecular basis of innate immune evasion by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an important consideration for designing the next wave of therapeutics. Here, we investigate the role of the nonstructural protein (NSP) 16 of SARS-CoV-2 in infection and pathogenesis. NSP16, a ribonucleoside 2’-O methyltransferase (MTase), catalyzes the transfer of a methyl group to mRNA as part of the capping process. Based on observations with other CoVs, we hypothesized that NSP16 2’-O MTase function protects SARS-CoV-2 from cap-sensing host restriction. Therefore, we engineered SARS-CoV-2 with a mutation that disrupts a conserved residue in the active site of NSP16. We subsequently show that this mutant is attenuated both in vitro and in vivo, using a hamster model of SARS-CoV-2 infection. Mechanistically, we confirm that the NSP16 mutant is more sensitive to type I interferon (IFN-I) in vitro. Furthermore, silencing IFIT1 or IFIT3, IFN-stimulated genes that sense a lack of 2’-O methylation, partially restores fitness to the NSP16 mutant. Conversely, overexpressing IFIT1 either alone or in combination with IFIT3 attenuates the NSP16 mutant relative to wild-type. Finally, we demonstrate that sinefungin, a MTase inhibitor that binds the catalytic site of NSP16, sensitizes wild-type SARS-CoV-2 to IFN-I treatment and attenuates viral replication in IFN-I competent cells. Overall, our findings highlight the importance of SARS-CoV-2 NSP16 to evading host innate immunity and suggest a possible target for future antiviral therapies.","abstract_html":"Understanding the molecular basis of innate immune evasion by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an important consideration for designing the next wave of therapeutics. Here, we investigate the role of the nonstructural protein (NSP) 16 of SARS-CoV-2 in infection and pathogenesis. NSP16, a ribonucleoside 2’-O methyltransferase (MTase), catalyzes the transfer of a methyl group to mRNA as part of the capping process. Based on observations with other CoVs, we hypothesized that NSP16 2’-O MTase function protects SARS-CoV-2 from cap-sensing host restriction. Therefore, we engineered SARS-CoV-2 with a mutation that disrupts a conserved residue in the active site of NSP16. We subsequently show that this mutant is attenuated both in vitro and in vivo, using a hamster model of SARS-CoV-2 infection. Mechanistically, we confirm that the NSP16 mutant is more sensitive to type I interferon (IFN-I) in vitro. Furthermore, silencing IFIT1 or IFIT3, IFN-stimulated genes that sense a lack of 2’-O methylation, partially restores fitness to the NSP16 mutant. Conversely, overexpressing IFIT1 either alone or in combination with IFIT3 attenuates the NSP16 mutant relative to wild-type. Finally, we demonstrate that sinefungin, a MTase inhibitor that binds the catalytic site of NSP16, sensitizes wild-type SARS-CoV-2 to IFN-I treatment and attenuates viral replication in IFN-I competent cells. Overall, our findings highlight the importance of SARS-CoV-2 NSP16 to evading host innate immunity and suggest a possible target for future antiviral therapies.","abstract_has_math":false,"creators":["Schindewolf, Craig Daniel 1987-"],"institution":"The University of Texas Medical Branch at Galveston","degree_name":"Microbiology and Immunology (Doctoral)","degree_level":null,"degree_discipline":"Microbiology and Immunology","degree_department":null,"school":null,"contributors":[],"advisors":["Menachery, Vineet"],"committee_chairs":[],"committee_members":["Endsley, Janice","Makino, Shinji","Rajsbaum, Ricardo","Daugherty, Matthew"],"year":2022,"date_issued":"2022-12-01T06:00:00.000Z","date_published":"2022-12-01T06:00:00.000Z","updated_at":"2026-07-24T05:51:09Z","subjects":[],"languages":["English"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/2152.3/11878","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Menachery, Vineet"]},{"key":"dc:contributor.committeemember","label":"Committee Member","values":["Endsley, Janice","Makino, Shinji","Rajsbaum, Ricardo","Daugherty, Matthew"]},{"key":"dc:creator","label":"Author","values":["Schindewolf, Craig Daniel 1987-"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2023-02-23T19:01:25Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2023-02-23T19:01:25Z"]},{"key":"dc:date.issued","label":"Date","values":["2022-12-01T06:00:00.000Z"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Microbiology and Immunology"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Microbiology and Immunology (Doctoral)"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["The University of Texas Medical Branch at Galveston"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["English"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/2152.3/11878"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Understanding the molecular basis of innate immune evasion by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an important consideration for designing the next wave of therapeutics. Here, we investigate the role of the nonstructural protein (NSP) 16 of SARS-CoV-2 in infection and pathogenesis. NSP16, a ribonucleoside 2’-O methyltransferase (MTase), catalyzes the transfer of a methyl group to mRNA as part of the capping process. Based on observations with other CoVs, we hypothesized that NSP16 2’-O MTase function protects SARS-CoV-2 from cap-sensing host restriction. Therefore, we engineered SARS-CoV-2 with a mutation that disrupts a conserved residue in the active site of NSP16. We subsequently show that this mutant is attenuated both in vitro and in vivo, using a hamster model of SARS-CoV-2 infection. Mechanistically, we confirm that the NSP16 mutant is more sensitive to type I interferon (IFN-I) in vitro. Furthermore, silencing IFIT1 or IFIT3, IFN-stimulated genes that sense a lack of 2’-O methylation, partially restores fitness to the NSP16 mutant. Conversely, overexpressing IFIT1 either alone or in combination with IFIT3 attenuates the NSP16 mutant relative to wild-type. Finally, we demonstrate that sinefungin, a MTase inhibitor that binds the catalytic site of NSP16, sensitizes wild-type SARS-CoV-2 to IFN-I treatment and attenuates viral replication in IFN-I competent cells. Overall, our findings highlight the importance of SARS-CoV-2 NSP16 to evading host innate immunity and suggest a possible target for future antiviral therapies."]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Subversion of innate antiviral immunity by SARS-CoV-2: An example from the study of nonstructural protein 16."]}]}],"canonical_facts":{"dc:contributor.advisor":["Menachery, Vineet"],"dc:contributor.committeemember":["Endsley, Janice","Makino, Shinji","Rajsbaum, Ricardo","Daugherty, Matthew"],"dc:creator":["Schindewolf, Craig Daniel 1987-"],"dc:date.accessioned":["2023-02-23T19:01:25Z"],"dc:date.available":["2023-02-23T19:01:25Z"],"dc:date.issued":["2022-12-01T06:00:00.000Z"],"dc:description.abstract":["Understanding the molecular basis of innate immune evasion by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an important consideration for designing the next wave of therapeutics. Here, we investigate the role of the nonstructural protein (NSP) 16 of SARS-CoV-2 in infection and pathogenesis. NSP16, a ribonucleoside 2’-O methyltransferase (MTase), catalyzes the transfer of a methyl group to mRNA as part of the capping process. Based on observations with other CoVs, we hypothesized that NSP16 2’-O MTase function protects SARS-CoV-2 from cap-sensing host restriction. Therefore, we engineered SARS-CoV-2 with a mutation that disrupts a conserved residue in the active site of NSP16. We subsequently show that this mutant is attenuated both in vitro and in vivo, using a hamster model of SARS-CoV-2 infection. Mechanistically, we confirm that the NSP16 mutant is more sensitive to type I interferon (IFN-I) in vitro. Furthermore, silencing IFIT1 or IFIT3, IFN-stimulated genes that sense a lack of 2’-O methylation, partially restores fitness to the NSP16 mutant. Conversely, overexpressing IFIT1 either alone or in combination with IFIT3 attenuates the NSP16 mutant relative to wild-type. Finally, we demonstrate that sinefungin, a MTase inhibitor that binds the catalytic site of NSP16, sensitizes wild-type SARS-CoV-2 to IFN-I treatment and attenuates viral replication in IFN-I competent cells. Overall, our findings highlight the importance of SARS-CoV-2 NSP16 to evading host innate immunity and suggest a possible target for future antiviral therapies."],"dc:format.mimetype":["application/pdf"],"dc:identifier.uri":["https://hdl.handle.net/2152.3/11878"],"dc:language.iso":["English"],"dc:title":["Subversion of innate antiviral immunity by SARS-CoV-2: An example from the study of nonstructural protein 16."],"dc:type":["Thesis"],"thesis:degree_discipline":["Microbiology and Immunology"],"thesis:degree_name":["Microbiology and Immunology (Doctoral)"],"thesis:institution_name":["The University of Texas Medical Branch at Galveston"]},"updated_at":"2026-07-24T05:51:09Z"}