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Article: A single residue substitution in the receptor-binding domain of H5N1 hemagglutinin is critical for packaging into pseudotyped lentiviral particles

TitleA single residue substitution in the receptor-binding domain of H5N1 hemagglutinin is critical for packaging into pseudotyped lentiviral particles
Authors
Issue Date2012
PublisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.action
Citation
PLoS One, 2012, v. 7 n. 11, p. e43596 How to Cite?
AbstractBACKGROUND: Serological studies for influenza infection and vaccine response often involve microneutralization and hemagglutination inhibition assays to evaluate neutralizing antibodies against human and avian influenza viruses, including H5N1. We have previously characterized lentiviral particles pseudotyped with H5-HA (H5pp) and validated an H5pp-based assay as a safe alternative for high-throughput serological studies in BSL-2 facilities. Here we show that H5-HAs from different clades do not always give rise to efficient production of H5pp and the underlying mechanisms are addressed. METHODOLOGY/FINDINGS: We have carried out mutational analysis to delineate the molecular determinants responsible for efficient packaging of HA from A/Cambodia/40808/2005 (H5Cam) and A/Anhui/1/2005 (H5Anh) into H5pp. Our results demonstrate that a single A134V mutation in the 130-loop of the receptor binding domain is sufficient to render H5Anh the ability to generate H5Anh-pp efficiently, whereas the reverse V134A mutation greatly hampers production of H5Cam-pp. Although protein expression in total cell lysates is similar for H5Anh and H5Cam, cell surface expression of H5Cam is detected at a significantly higher level than that of H5Anh. We further demonstrate by several independent lines of evidence that the behaviour of H5Anh can be explained by a stronger binding to sialic acid receptors implicating residue 134. CONCLUSIONS: We have identified a single A134V mutation as the molecular determinant in H5-HA for efficient incorporation into H5pp envelope and delineated the underlying mechanism. The reduced binding to sialic acid receptors as a result of the A134V mutation not only exerts a critical influence in pseudotyping efficiency of H5-HA, but has also an impact at the whole virus level. Because A134V substitution has been reported as a naturally occurring mutation in human host, our results may have implications for the understanding of human host adaptation of avian influenza H5N1 viruses.
Persistent Identifierhttp://hdl.handle.net/10722/191973
ISSN
2015 Impact Factor: 3.057
2015 SCImago Journal Rankings: 1.395
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorTang, Den_US
dc.contributor.authorLam, YMen_US
dc.contributor.authorSiu, YLen_US
dc.contributor.authorLam, CHen_US
dc.contributor.authorChu, SLen_US
dc.contributor.authorPeiris, JSMen_US
dc.contributor.authorBuchy, Pen_US
dc.contributor.authorNai, Ben_US
dc.contributor.authorBruzzone, Ren_US
dc.date.accessioned2013-10-15T07:44:01Z-
dc.date.available2013-10-15T07:44:01Z-
dc.date.issued2012en_US
dc.identifier.citationPLoS One, 2012, v. 7 n. 11, p. e43596en_US
dc.identifier.issn1932-6203-
dc.identifier.urihttp://hdl.handle.net/10722/191973-
dc.description.abstractBACKGROUND: Serological studies for influenza infection and vaccine response often involve microneutralization and hemagglutination inhibition assays to evaluate neutralizing antibodies against human and avian influenza viruses, including H5N1. We have previously characterized lentiviral particles pseudotyped with H5-HA (H5pp) and validated an H5pp-based assay as a safe alternative for high-throughput serological studies in BSL-2 facilities. Here we show that H5-HAs from different clades do not always give rise to efficient production of H5pp and the underlying mechanisms are addressed. METHODOLOGY/FINDINGS: We have carried out mutational analysis to delineate the molecular determinants responsible for efficient packaging of HA from A/Cambodia/40808/2005 (H5Cam) and A/Anhui/1/2005 (H5Anh) into H5pp. Our results demonstrate that a single A134V mutation in the 130-loop of the receptor binding domain is sufficient to render H5Anh the ability to generate H5Anh-pp efficiently, whereas the reverse V134A mutation greatly hampers production of H5Cam-pp. Although protein expression in total cell lysates is similar for H5Anh and H5Cam, cell surface expression of H5Cam is detected at a significantly higher level than that of H5Anh. We further demonstrate by several independent lines of evidence that the behaviour of H5Anh can be explained by a stronger binding to sialic acid receptors implicating residue 134. CONCLUSIONS: We have identified a single A134V mutation as the molecular determinant in H5-HA for efficient incorporation into H5pp envelope and delineated the underlying mechanism. The reduced binding to sialic acid receptors as a result of the A134V mutation not only exerts a critical influence in pseudotyping efficiency of H5-HA, but has also an impact at the whole virus level. Because A134V substitution has been reported as a naturally occurring mutation in human host, our results may have implications for the understanding of human host adaptation of avian influenza H5N1 viruses.-
dc.languageengen_US
dc.publisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.action-
dc.relation.ispartofPLoS Oneen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subject.meshHemagglutinin Glycoproteins, Influenza Virus - chemistry - genetics-
dc.subject.meshInfluenza A Virus, H5N1 Subtype - chemistry-
dc.subject.meshInfluenza Vaccines - chemistry-
dc.subject.meshLentivirus - metabolism-
dc.subject.meshMadin Darby Canine Kidney Cells-
dc.titleA single residue substitution in the receptor-binding domain of H5N1 hemagglutinin is critical for packaging into pseudotyped lentiviral particlesen_US
dc.typeArticleen_US
dc.identifier.emailrTang, D: djtang@graduate.hku.hken_US
dc.identifier.emailSiu, YL: ylsiu@hkucc.hku.hken_US
dc.identifier.emailPeiris, JSM: malik@hkucc.hku.hken_US
dc.identifier.emailBruzzone, R: bruzzone@hkucc.hku.hken_US
dc.identifier.authorityPeiris, JSM=rp00410en_US
dc.identifier.authorityBruzzone, R=rp01442en_US
dc.description.naturepublished_or_final_version-
dc.identifier.doi10.1371/journal.pone.0043596-
dc.identifier.pmid23133587-
dc.identifier.pmcidPMC3487904-
dc.identifier.hkuros226147en_US
dc.identifier.volume7en_US
dc.identifier.issue11en_US
dc.identifier.spagee43596en_US
dc.identifier.epagee43596en_US
dc.identifier.isiWOS:000310702400001-
dc.publisher.placeUnited States-

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