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Article: Single amino acid substitutions in the severe acute respiratory syndrome coronavirus spike glycoprotein determine viral entry and immunogenicity of a major neutralizing domain

TitleSingle amino acid substitutions in the severe acute respiratory syndrome coronavirus spike glycoprotein determine viral entry and immunogenicity of a major neutralizing domain
Authors
Issue Date2005
PublisherAmerican Society for Microbiology. The Journal's web site is located at http://jvi.asm.org/
Citation
Journal Of Virology, 2005, v. 79 n. 18, p. 11638-11646 How to Cite?
AbstractNeutralizing antibodies (NAbs) against severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) spike (S) glycoprotein confer protection to animals experimentally infected with the pathogenic virus. We and others previously demonstrated that a major mechanism for neutralizing SARS-CoV was through blocking the interaction between the S glycoprotein and the cellular receptor angiotensin-converting enzyme 2 (ACE2). In this study, we used in vivo electroporation DNA immunization and a pseudovirus-based assay to functionally evaluate immunogenicity and viral entry. We characterized the neutralization and viral entry determinants within the ACE2-binding domain of the S glycoprotein. The deletion of a positively charged region SΔ(422-463) abolished the capacity of the S glycoprotein to induce NAbs in mice vaccinated by in vivo DNA electroporation. Moreover, the SΔ(422-463) pseadovirus was unable to infect HEK293T-ACE2 cells. To determine the specific residues that contribute to related phenotypes, we replaced eight basic amino acids with alanine. We found that a single amino acid substitution (R441A) in the full-length S DNA vaccine failed to induce NAbs and abolished viral entry when pseudoviruses were generated. However, another substitution (R453A) abolished viral entry while retaining the capacity for inducing NAbs. The difference between R441A and R453A suggests that the determinants for immunogenicity and viral entry may not be identical. Our findings provide direct evidence that these basic residues are essential for immunogenicity of the major neutralizing domain and for viral entry. Our data have implications for the rational design of vaccine and antiviral agents as well as for understanding viral tropism. Copyright © 2005, American Society for Microbiology. All Rights Reserved.
Persistent Identifierhttp://hdl.handle.net/10722/157416
ISSN
2015 Impact Factor: 4.606
2015 SCImago Journal Rankings: 3.347
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorYi, CEen_US
dc.contributor.authorBa, Len_US
dc.contributor.authorZhang, Len_US
dc.contributor.authorHo, DDen_US
dc.contributor.authorChen, Zen_US
dc.date.accessioned2012-08-08T08:49:46Z-
dc.date.available2012-08-08T08:49:46Z-
dc.date.issued2005en_US
dc.identifier.citationJournal Of Virology, 2005, v. 79 n. 18, p. 11638-11646en_US
dc.identifier.issn0022-538Xen_US
dc.identifier.urihttp://hdl.handle.net/10722/157416-
dc.description.abstractNeutralizing antibodies (NAbs) against severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) spike (S) glycoprotein confer protection to animals experimentally infected with the pathogenic virus. We and others previously demonstrated that a major mechanism for neutralizing SARS-CoV was through blocking the interaction between the S glycoprotein and the cellular receptor angiotensin-converting enzyme 2 (ACE2). In this study, we used in vivo electroporation DNA immunization and a pseudovirus-based assay to functionally evaluate immunogenicity and viral entry. We characterized the neutralization and viral entry determinants within the ACE2-binding domain of the S glycoprotein. The deletion of a positively charged region SΔ(422-463) abolished the capacity of the S glycoprotein to induce NAbs in mice vaccinated by in vivo DNA electroporation. Moreover, the SΔ(422-463) pseadovirus was unable to infect HEK293T-ACE2 cells. To determine the specific residues that contribute to related phenotypes, we replaced eight basic amino acids with alanine. We found that a single amino acid substitution (R441A) in the full-length S DNA vaccine failed to induce NAbs and abolished viral entry when pseudoviruses were generated. However, another substitution (R453A) abolished viral entry while retaining the capacity for inducing NAbs. The difference between R441A and R453A suggests that the determinants for immunogenicity and viral entry may not be identical. Our findings provide direct evidence that these basic residues are essential for immunogenicity of the major neutralizing domain and for viral entry. Our data have implications for the rational design of vaccine and antiviral agents as well as for understanding viral tropism. Copyright © 2005, American Society for Microbiology. All Rights Reserved.en_US
dc.languageengen_US
dc.publisherAmerican Society for Microbiology. The Journal's web site is located at http://jvi.asm.org/en_US
dc.relation.ispartofJournal of Virologyen_US
dc.subject.meshAmino Acid Sequenceen_US
dc.subject.meshAmino Acid Substitutionen_US
dc.subject.meshAnimalsen_US
dc.subject.meshAntibodies, Viral - Biosynthesisen_US
dc.subject.meshAntigens, Viral - Chemistry - Geneticsen_US
dc.subject.meshCell Lineen_US
dc.subject.meshFemaleen_US
dc.subject.meshHumansen_US
dc.subject.meshMembrane Glycoproteins - Chemistry - Genetics - Immunology - Physiologyen_US
dc.subject.meshMiceen_US
dc.subject.meshMice, Inbred Balb Cen_US
dc.subject.meshMicroscopy, Electronen_US
dc.subject.meshMutagenesis, Site-Directeden_US
dc.subject.meshNeutralization Testsen_US
dc.subject.meshProtein Structure, Tertiaryen_US
dc.subject.meshSars Virus - Genetics - Immunology - Pathogenicity - Physiologyen_US
dc.subject.meshSequence Deletionen_US
dc.subject.meshVaccines, Dna - Administration & Dosage - Geneticsen_US
dc.subject.meshViral Envelope Proteins - Chemistry - Genetics - Immunology - Physiologyen_US
dc.subject.meshViral Vaccines - Administration & Dosage - Geneticsen_US
dc.subject.meshVirulence - Genetics - Immunology - Physiologyen_US
dc.titleSingle amino acid substitutions in the severe acute respiratory syndrome coronavirus spike glycoprotein determine viral entry and immunogenicity of a major neutralizing domainen_US
dc.typeArticleen_US
dc.identifier.emailChen, Z:zchenai@hkucc.hku.hken_US
dc.identifier.authorityChen, Z=rp00243en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1128/JVI.79.18.11638-11646.2005en_US
dc.identifier.pmid16140741-
dc.identifier.scopuseid_2-s2.0-24644508802en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-24644508802&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume79en_US
dc.identifier.issue18en_US
dc.identifier.spage11638en_US
dc.identifier.epage11646en_US
dc.identifier.isiWOS:000231633900010-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridYi, CE=8557032800en_US
dc.identifier.scopusauthoridBa, L=8557032300en_US
dc.identifier.scopusauthoridZhang, L=8783285300en_US
dc.identifier.scopusauthoridHo, DD=7402971998en_US
dc.identifier.scopusauthoridChen, Z=35271180800en_US

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