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dc.contributor.authorJain, Surbhi
dc.contributor.authorCullen, Lori McGinnes
dc.contributor.authorMorrison, Trudy G.
dc.date2022-08-11T08:08:59.000
dc.date.accessioned2022-08-23T16:14:40Z
dc.date.available2022-08-23T16:14:40Z
dc.date.issued2006-12-08
dc.date.submitted2008-10-09
dc.identifier.citationJ Virol. 2007 Mar;81(5):2328-39. Epub 2006 Dec 6. <a href="http://dx.doi.org/10.1128/JVI.01940-06 ">Link to article on publisher's site</a>
dc.identifier.issn0022-538X (Print)
dc.identifier.doi10.1128/JVI.01940-06
dc.identifier.pmid17151113
dc.identifier.urihttp://hdl.handle.net/20.500.14038/33914
dc.description.abstractNewcastle disease virus (NDV), an avian paramyxovirus, initiates infection with attachment of the viral hemagglutinin-neuraminidase (HN) protein to sialic acid-containing receptors, followed by fusion of viral and cell membranes, which is mediated by the fusion (F) protein. Like all class 1 viral fusion proteins, the paramyxovirus F protein is thought to undergo dramatic conformational changes upon activation. How the F protein accomplishes extensive conformational rearrangements is unclear. Since several viral fusion proteins undergo disulfide bond rearrangement during entry, we asked if similar rearrangements occur in NDV proteins during entry. We found that inhibitors of cell surface thiol/disulfide isomerase activity--5'5-dithio-bis(2-nitrobenzoic acid) (DTNB), bacitracin, and anti-protein disulfide isomerase antibody--inhibited cell-cell fusion and virus entry but had no effect on cell viability, glycoprotein surface expression, or HN protein attachment or neuraminidase activities. These inhibitors altered the conformation of surface-expressed F protein, as detected by conformation-sensitive antibodies. Using biotin maleimide (MPB), a reagent that binds to free thiols, free thiols were detected on surface-expressed F protein, but not HN protein. The inhibitors DTNB and bacitracin blocked the detection of these free thiols. Furthermore, MPB binding inhibited cell-cell fusion. Taken together, our results suggest that one or several disulfide bonds in cell surface F protein are reduced by the protein disulfide isomerase family of isomerases and that F protein exists as a mixture of oxidized and reduced forms. In the presence of HN protein, only the reduced form may proceed to refold into additional intermediates, leading to the fusion of membranes.
dc.language.isoen_US
dc.relation<a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=17151113&dopt=Abstract">Link to article in PubMed</a>
dc.relation.urlhttp://dx.doi.org/10.1128/JVI.01940-06
dc.subjectAnimals; Bacitracin; COS Cells; Cercopithecus aethiops; Disulfides; Dithionitrobenzoic Acid; Enzyme Inhibitors; HN Protein; Membrane Fusion; Models, Biological; Newcastle disease virus; Protein Disulfide-Isomerase; Recombinant Proteins; Sulfhydryl Compounds; Viral Fusion Proteins; Virus Attachment
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleThiol/disulfide exchange is required for membrane fusion directed by the Newcastle disease virus fusion protein
dc.typeJournal Article
dc.source.journaltitleJournal of virology
dc.source.volume81
dc.source.issue5
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/gsbs_sp/569
dc.identifier.contextkey646754
html.description.abstract<p>Newcastle disease virus (NDV), an avian paramyxovirus, initiates infection with attachment of the viral hemagglutinin-neuraminidase (HN) protein to sialic acid-containing receptors, followed by fusion of viral and cell membranes, which is mediated by the fusion (F) protein. Like all class 1 viral fusion proteins, the paramyxovirus F protein is thought to undergo dramatic conformational changes upon activation. How the F protein accomplishes extensive conformational rearrangements is unclear. Since several viral fusion proteins undergo disulfide bond rearrangement during entry, we asked if similar rearrangements occur in NDV proteins during entry. We found that inhibitors of cell surface thiol/disulfide isomerase activity--5'5-dithio-bis(2-nitrobenzoic acid) (DTNB), bacitracin, and anti-protein disulfide isomerase antibody--inhibited cell-cell fusion and virus entry but had no effect on cell viability, glycoprotein surface expression, or HN protein attachment or neuraminidase activities. These inhibitors altered the conformation of surface-expressed F protein, as detected by conformation-sensitive antibodies. Using biotin maleimide (MPB), a reagent that binds to free thiols, free thiols were detected on surface-expressed F protein, but not HN protein. The inhibitors DTNB and bacitracin blocked the detection of these free thiols. Furthermore, MPB binding inhibited cell-cell fusion. Taken together, our results suggest that one or several disulfide bonds in cell surface F protein are reduced by the protein disulfide isomerase family of isomerases and that F protein exists as a mixture of oxidized and reduced forms. In the presence of HN protein, only the reduced form may proceed to refold into additional intermediates, leading to the fusion of membranes.</p>
dc.identifier.submissionpathgsbs_sp/569
dc.contributor.departmentDepartment of Molecular Genetics and Microbiology
dc.contributor.departmentGraduate School of Biomedical Sciences
dc.source.pages2328-39


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