{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/104997"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/104997","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Innate immune modulation and pathogenic basis for porcine reproductive and respiratory syndrome virus","abstract":"Porcine reproductive and respiratory syndrome virus causes PRRS which is one of the most economically significant diseases in the swine industry worldwide. PRRSV has the ability to suppress the type I interferon induction to facilitate its survival during infection, and the nsp1 protein of PRRSV has been identified as the potent IFN antagonist. The nsp1β subunit of nsp1 blocks the host mRNA nuclear export and this is one of the viral mechanisms to inhibit host antiviral protein translation. The functional motif for both IFN suppression and host mRNA nuclear retention were identified in nsp1β, and mutations in the motif results in the IFN-suppression-negative and host-mRNA nuclear retention-negative. To examine the pathogenic consequence of these functions in the natural host animals, two mutant PRRS viruses (vL126A and vL135A) were generated using infectious clones. These mutant viruses retained the infectivity, and their phenotype was IFN suppression-negative and host mRNA nuclear retention-negative. Pigs infected with vL126A or vL135A exhibited milder severe clinical signs with lower viral titers and shorter duration of viremia when compared to those of WT-infected pigs. The levels of PRRSV-specific antibody remained comparable in all infected groups but the neutralizing antibody titers were high in vL126A-infected or vL135A-infected animals. The IFN-α concentration was also high in pigs infected with the mutant PRRSV. Subsequently, the nsp1β mediated inhibitory mechanism was investigated. Nsp1β was found to specifically interact with nucleoporin 62 (Nup62). Nup62 is one of the major components of the nuclear pore complex (NPC) that form channels spanning the double lipid bilayer of the nuclear envelope for nucleocytoplasmic transport of cellular molecules including host mRNAs. A region representing the C-terminal 328-522 residues of Nup62 was identified as the binding domain to nsp1β. Mutational studies revealed that leucine 126 of nsp1β was the critical residue for Nup62 interaction. Nsp1β L126A did not bind to Nup62, and host mRNA nuclear export occurred normally. In nsp1β-overexpressing cells or siRNA-mediated Nup62 knock-down cells, viral growth was improved. This is likely attributed to nsp1β-mediated viral protein production enhancement and reduction of antiviral proteins in the cytoplasm. One of the mechanisms that PRRSV causes respiratory disease is the onset of inflammation at the site of infection. PRRSV activates NF-kB during infection, and the nucleocapsid (N) protein has been identified as the major NF-kB activator. The protein inhibitor of activated STAT1 (PIAS1) was identified as a molecular partner of N. PIAS1 binds to RelA (p65) and prevents NF-kB activation by interfering p65-DNA binding in the nucleus and thus functions as a negative regulator of NF-kB. The binding domain of PIAS1 was mapped to the N-terminal domain, and this domain was sufficient to bind to p65 and prevent its binding to the kB site, demonstrating the competitive binding between N-PIAS1 and NF-kB-PIAS1. For N, the region between 37 and 72 amino acids was found to interact with PIAS1, and this region overlapped the nuclear localization signal (NLS) of N. A nuclear localization signal (NLS) knock-out mutant N did not activate NF-κB, and this is likely due to the lack of its interaction with PIAS1 in the nucleus, demonstrating the positive correlation between the binding of N to PIAS1 and the NF-κB activation. In conclusion, the current study dissects the molecular basis of innate immune suppression and NF-kB activation of PRRSV and identifies the leucine 126 of nsp1β and NLS of N are essential residues. The innate immune modulation and NF-kB activation together delineate the pathogenic basis of PRRSV.","abstract_html":"Porcine reproductive and respiratory syndrome virus causes PRRS which is one of the most economically significant diseases in the swine industry worldwide. PRRSV has the ability to suppress the type I interferon induction to facilitate its survival during infection, and the nsp1 protein of PRRSV has been identified as the potent IFN antagonist. The nsp1β subunit of nsp1 blocks the host mRNA nuclear export and this is one of the viral mechanisms to inhibit host antiviral protein translation. The functional motif for both IFN suppression and host mRNA nuclear retention were identified in nsp1β, and mutations in the motif results in the IFN-suppression-negative and host-mRNA nuclear retention-negative. To examine the pathogenic consequence of these functions in the natural host animals, two mutant PRRS viruses (vL126A and vL135A) were generated using infectious clones. These mutant viruses retained the infectivity, and their phenotype was IFN suppression-negative and host mRNA nuclear retention-negative. Pigs infected with vL126A or vL135A exhibited milder severe clinical signs with lower viral titers and shorter duration of viremia when compared to those of WT-infected pigs. The levels of PRRSV-specific antibody remained comparable in all infected groups but the neutralizing antibody titers were high in vL126A-infected or vL135A-infected animals. The IFN-α concentration was also high in pigs infected with the mutant PRRSV. Subsequently, the nsp1β mediated inhibitory mechanism was investigated. Nsp1β was found to specifically interact with nucleoporin 62 (Nup62). Nup62 is one of the major components of the nuclear pore complex (NPC) that form channels spanning the double lipid bilayer of the nuclear envelope for nucleocytoplasmic transport of cellular molecules including host mRNAs. A region representing the C-terminal 328-522 residues of Nup62 was identified as the binding domain to nsp1β. Mutational studies revealed that leucine 126 of nsp1β was the critical residue for Nup62 interaction. Nsp1β L126A did not bind to Nup62, and host mRNA nuclear export occurred normally. In nsp1β-overexpressing cells or siRNA-mediated Nup62 knock-down cells, viral growth was improved. This is likely attributed to nsp1β-mediated viral protein production enhancement and reduction of antiviral proteins in the cytoplasm. One of the mechanisms that PRRSV causes respiratory disease is the onset of inflammation at the site of infection. PRRSV activates NF-kB during infection, and the nucleocapsid (N) protein has been identified as the major NF-kB activator. The protein inhibitor of activated STAT1 (PIAS1) was identified as a molecular partner of N. PIAS1 binds to RelA (p65) and prevents NF-kB activation by interfering p65-DNA binding in the nucleus and thus functions as a negative regulator of NF-kB. The binding domain of PIAS1 was mapped to the N-terminal domain, and this domain was sufficient to bind to p65 and prevent its binding to the kB site, demonstrating the competitive binding between N-PIAS1 and NF-kB-PIAS1. For N, the region between 37 and 72 amino acids was found to interact with PIAS1, and this region overlapped the nuclear localization signal (NLS) of N. A nuclear localization signal (NLS) knock-out mutant N did not activate NF-κB, and this is likely due to the lack of its interaction with PIAS1 in the nucleus, demonstrating the positive correlation between the binding of N to PIAS1 and the NF-κB activation. In conclusion, the current study dissects the molecular basis of innate immune suppression and NF-kB activation of PRRSV and identifies the leucine 126 of nsp1β and NLS of N are essential residues. The innate immune modulation and NF-kB activation together delineate the pathogenic basis of PRRSV.","abstract_has_math":false,"creators":["Ke, Hanzhong"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"VMS - Pathobiology","degree_department":null,"school":null,"contributors":["Yoo, Dongwan","Rock, Daniel L","Zuckermann, Federico A.","Brooke, Christopher Byron"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2019,"date_issued":"2019-08-23T20:35:48Z","date_published":"2019-08-23T20:35:48Z","updated_at":"2026-07-22T22:24:44Z","subjects":["Porcine reproductive and respiratory syndrome virus","PRRSV","Innate immune","NF-kB","pathogenesis","SAP motif","Nucleoporin 62","Nup62","PIAS1","type I IFN","pro-inflammatory cytokines"],"languages":["en"],"rights":["Copyright 2019 Hanzhong Ke"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/104997","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Yoo, Dongwan","Rock, Daniel L","Zuckermann, Federico A.","Brooke, Christopher Byron"]},{"key":"dc:creator","label":"Author","values":["Ke, Hanzhong"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2019-08-23T20:35:48Z","2021-08-24T09:15:28Z","2019-04-09","2019-05"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["VMS - Pathobiology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Porcine reproductive and respiratory syndrome virus","PRRSV","Innate immune","NF-kB","pathogenesis","SAP motif","Nucleoporin 62","Nup62","PIAS1","type I IFN","pro-inflammatory cytokines"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2019 Hanzhong Ke"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/104997"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Porcine reproductive and respiratory syndrome virus causes PRRS which is one of the most economically significant diseases in the swine industry worldwide. PRRSV has the ability to suppress the type I interferon induction to facilitate its survival during infection, and the nsp1 protein of PRRSV has been identified as the potent IFN antagonist. The nsp1β subunit of nsp1 blocks the host mRNA nuclear export and this is one of the viral mechanisms to inhibit host antiviral protein translation. The functional motif for both IFN suppression and host mRNA nuclear retention were identified in nsp1β, and mutations in the motif results in the IFN-suppression-negative and host-mRNA nuclear retention-negative. To examine the pathogenic consequence of these functions in the natural host animals, two mutant PRRS viruses (vL126A and vL135A) were generated using infectious clones. These mutant viruses retained the infectivity, and their phenotype was IFN suppression-negative and host mRNA nuclear retention-negative. Pigs infected with vL126A or vL135A exhibited milder severe clinical signs with lower viral titers and shorter duration of viremia when compared to those of WT-infected pigs. The levels of PRRSV-specific antibody remained comparable in all infected groups but the neutralizing antibody titers were high in vL126A-infected or vL135A-infected animals. The IFN-α concentration was also high in pigs infected with the mutant PRRSV. Subsequently, the nsp1β mediated inhibitory mechanism was investigated. Nsp1β was found to specifically interact with nucleoporin 62 (Nup62). Nup62 is one of the major components of the nuclear pore complex (NPC) that form channels spanning the double lipid bilayer of the nuclear envelope for nucleocytoplasmic transport of cellular molecules including host mRNAs. A region representing the C-terminal 328-522 residues of Nup62 was identified as the binding domain to nsp1β. Mutational studies revealed that leucine 126 of nsp1β was the critical residue for Nup62 interaction. Nsp1β L126A did not bind to Nup62, and host mRNA nuclear export occurred normally. In nsp1β-overexpressing cells or siRNA-mediated Nup62 knock-down cells, viral growth was improved. This is likely attributed to nsp1β-mediated viral protein production enhancement and reduction of antiviral proteins in the cytoplasm. One of the mechanisms that PRRSV causes respiratory disease is the onset of inflammation at the site of infection. PRRSV activates NF-kB during infection, and the nucleocapsid (N) protein has been identified as the major NF-kB activator. The protein inhibitor of activated STAT1 (PIAS1) was identified as a molecular partner of N. PIAS1 binds to RelA (p65) and prevents NF-kB activation by interfering p65-DNA binding in the nucleus and thus functions as a negative regulator of NF-kB. The binding domain of PIAS1 was mapped to the N-terminal domain, and this domain was sufficient to bind to p65 and prevent its binding to the kB site, demonstrating the competitive binding between N-PIAS1 and NF-kB-PIAS1. For N, the region between 37 and 72 amino acids was found to interact with PIAS1, and this region overlapped the nuclear localization signal (NLS) of N. A nuclear localization signal (NLS) knock-out mutant N did not activate NF-κB, and this is likely due to the lack of its interaction with PIAS1 in the nucleus, demonstrating the positive correlation between the binding of N to PIAS1 and the NF-κB activation. In conclusion, the current study dissects the molecular basis of innate immune suppression and NF-kB activation of PRRSV and identifies the leucine 126 of nsp1β and NLS of N are essential residues. The innate immune modulation and NF-kB activation together delineate the pathogenic basis of PRRSV.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2021-05-01","The student, Hanzhong Ke, accepted the attached license on 2019-04-09 at 14:13.","The student, Hanzhong Ke, submitted this Dissertation for approval on 2019-04-09 at 14:26.","This Dissertation was approved for publication on 2019-04-09 at 14:42.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13537 on 2019-08-22 at 15:05:39","Made available in DSpace on 2019-08-23T20:35:48Z (GMT). No. of bitstreams: 2 KE-DISSERTATION-2019.pdf: 13693069 bytes, checksum: c81a69abc29007f9a246331a5d623cec (MD5) LICENSE.txt: 4208 bytes, checksum: 22575214c4077653eb8a3a1041000b03 (MD5) Previous issue date: 2019-04-09","Embargo set by: Seth Robbins for item 112116 Lift date: 2021-08-23T20:36:18Z Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","U of I Only Restriction Lifted for Item 112116 on 2021-08-24T09:15:28Z."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Innate immune modulation and pathogenic basis for porcine reproductive and respiratory syndrome virus"]}]}],"canonical_facts":{"dc:contributor":["Yoo, Dongwan","Rock, Daniel L","Zuckermann, Federico A.","Brooke, Christopher Byron"],"dc:creator":["Ke, Hanzhong"],"dc:date":["2019-08-23T20:35:48Z","2021-08-24T09:15:28Z","2019-04-09","2019-05"],"dc:description":["Porcine reproductive and respiratory syndrome virus causes PRRS which is one of the most economically significant diseases in the swine industry worldwide. PRRSV has the ability to suppress the type I interferon induction to facilitate its survival during infection, and the nsp1 protein of PRRSV has been identified as the potent IFN antagonist. The nsp1β subunit of nsp1 blocks the host mRNA nuclear export and this is one of the viral mechanisms to inhibit host antiviral protein translation. The functional motif for both IFN suppression and host mRNA nuclear retention were identified in nsp1β, and mutations in the motif results in the IFN-suppression-negative and host-mRNA nuclear retention-negative. To examine the pathogenic consequence of these functions in the natural host animals, two mutant PRRS viruses (vL126A and vL135A) were generated using infectious clones. These mutant viruses retained the infectivity, and their phenotype was IFN suppression-negative and host mRNA nuclear retention-negative. Pigs infected with vL126A or vL135A exhibited milder severe clinical signs with lower viral titers and shorter duration of viremia when compared to those of WT-infected pigs. The levels of PRRSV-specific antibody remained comparable in all infected groups but the neutralizing antibody titers were high in vL126A-infected or vL135A-infected animals. The IFN-α concentration was also high in pigs infected with the mutant PRRSV. Subsequently, the nsp1β mediated inhibitory mechanism was investigated. Nsp1β was found to specifically interact with nucleoporin 62 (Nup62). Nup62 is one of the major components of the nuclear pore complex (NPC) that form channels spanning the double lipid bilayer of the nuclear envelope for nucleocytoplasmic transport of cellular molecules including host mRNAs. A region representing the C-terminal 328-522 residues of Nup62 was identified as the binding domain to nsp1β. Mutational studies revealed that leucine 126 of nsp1β was the critical residue for Nup62 interaction. Nsp1β L126A did not bind to Nup62, and host mRNA nuclear export occurred normally. In nsp1β-overexpressing cells or siRNA-mediated Nup62 knock-down cells, viral growth was improved. This is likely attributed to nsp1β-mediated viral protein production enhancement and reduction of antiviral proteins in the cytoplasm. One of the mechanisms that PRRSV causes respiratory disease is the onset of inflammation at the site of infection. PRRSV activates NF-kB during infection, and the nucleocapsid (N) protein has been identified as the major NF-kB activator. The protein inhibitor of activated STAT1 (PIAS1) was identified as a molecular partner of N. PIAS1 binds to RelA (p65) and prevents NF-kB activation by interfering p65-DNA binding in the nucleus and thus functions as a negative regulator of NF-kB. The binding domain of PIAS1 was mapped to the N-terminal domain, and this domain was sufficient to bind to p65 and prevent its binding to the kB site, demonstrating the competitive binding between N-PIAS1 and NF-kB-PIAS1. For N, the region between 37 and 72 amino acids was found to interact with PIAS1, and this region overlapped the nuclear localization signal (NLS) of N. A nuclear localization signal (NLS) knock-out mutant N did not activate NF-κB, and this is likely due to the lack of its interaction with PIAS1 in the nucleus, demonstrating the positive correlation between the binding of N to PIAS1 and the NF-κB activation. In conclusion, the current study dissects the molecular basis of innate immune suppression and NF-kB activation of PRRSV and identifies the leucine 126 of nsp1β and NLS of N are essential residues. The innate immune modulation and NF-kB activation together delineate the pathogenic basis of PRRSV.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2021-05-01","The student, Hanzhong Ke, accepted the attached license on 2019-04-09 at 14:13.","The student, Hanzhong Ke, submitted this Dissertation for approval on 2019-04-09 at 14:26.","This Dissertation was approved for publication on 2019-04-09 at 14:42.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13537 on 2019-08-22 at 15:05:39","Made available in DSpace on 2019-08-23T20:35:48Z (GMT). No. of bitstreams: 2 KE-DISSERTATION-2019.pdf: 13693069 bytes, checksum: c81a69abc29007f9a246331a5d623cec (MD5) LICENSE.txt: 4208 bytes, checksum: 22575214c4077653eb8a3a1041000b03 (MD5) Previous issue date: 2019-04-09","Embargo set by: Seth Robbins for item 112116 Lift date: 2021-08-23T20:36:18Z Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","U of I Only Restriction Lifted for Item 112116 on 2021-08-24T09:15:28Z."],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/2142/104997"],"dc:language":["en"],"dc:rights":["Copyright 2019 Hanzhong Ke"],"dc:subject":["Porcine reproductive and respiratory syndrome virus","PRRSV","Innate immune","NF-kB","pathogenesis","SAP motif","Nucleoporin 62","Nup62","PIAS1","type I IFN","pro-inflammatory cytokines"],"dc:title":["Innate immune modulation and pathogenic basis for porcine reproductive and respiratory syndrome virus"],"dc:type":["text"],"thesis:degree_discipline":["VMS - Pathobiology"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:24:44Z"}