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Article: A viral deubiquitylating enzyme restores dislocation of substrates from the endoplasmic reticulum (ER) in semi-intact cells

TitleA viral deubiquitylating enzyme restores dislocation of substrates from the endoplasmic reticulum (ER) in semi-intact cells
Authors
Issue Date2012
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
Citation
Journal Of Biological Chemistry, 2012, v. 287 n. 28, p. 23594-23603 How to Cite?
AbstractTerminally misfolded glycoproteins are ejected from the endoplasmic reticulum (ER) to the cytosol and are destroyed by the ubiquitin proteasome system. A dominant negative version of the deubiquitylating enzyme Yod1 (Yod1C160S) causes accumulation of dislocation substrates in the ER. Failure to remove ubiquitin from the dislocation substrate might therefore stall the reaction at the exit site from the ER. We hypothesized that addition of a promiscuous deubiquitylase should overcome this blockade and restore dislocation. We monitored ER-to-cytosol transport of misfolded proteins in cells permeabilized at high cell density by perfringolysin O, a pore-forming cytolysin. This method allows ready access of otherwise impermeant reagents to the intracellular milieu with minimal dilution of cytoplasmic components. We show that addition of the purified Epstein-Barr virus deubiquitylase to semi-intact cells indeed initiates dislocation of a stalled substrate intermediate, resulting in stabilization of substrates in the cytosol. Our data provide new mechanistic insight in the dislocation reaction and support a model where failure to deubiquitylate an ER-resident protein occludes the dislocon and causes upstream misfolded intermediates to accumulate. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.
Persistent Identifierhttp://hdl.handle.net/10722/188685
ISSN
2015 Impact Factor: 4.258
2015 SCImago Journal Rankings: 3.151
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorSanyal, Sen_US
dc.contributor.authorClaessen, JHLen_US
dc.contributor.authorPloegh, HLen_US
dc.date.accessioned2013-09-03T04:12:45Z-
dc.date.available2013-09-03T04:12:45Z-
dc.date.issued2012en_US
dc.identifier.citationJournal Of Biological Chemistry, 2012, v. 287 n. 28, p. 23594-23603en_US
dc.identifier.issn0021-9258en_US
dc.identifier.urihttp://hdl.handle.net/10722/188685-
dc.description.abstractTerminally misfolded glycoproteins are ejected from the endoplasmic reticulum (ER) to the cytosol and are destroyed by the ubiquitin proteasome system. A dominant negative version of the deubiquitylating enzyme Yod1 (Yod1C160S) causes accumulation of dislocation substrates in the ER. Failure to remove ubiquitin from the dislocation substrate might therefore stall the reaction at the exit site from the ER. We hypothesized that addition of a promiscuous deubiquitylase should overcome this blockade and restore dislocation. We monitored ER-to-cytosol transport of misfolded proteins in cells permeabilized at high cell density by perfringolysin O, a pore-forming cytolysin. This method allows ready access of otherwise impermeant reagents to the intracellular milieu with minimal dilution of cytoplasmic components. We show that addition of the purified Epstein-Barr virus deubiquitylase to semi-intact cells indeed initiates dislocation of a stalled substrate intermediate, resulting in stabilization of substrates in the cytosol. Our data provide new mechanistic insight in the dislocation reaction and support a model where failure to deubiquitylate an ER-resident protein occludes the dislocon and causes upstream misfolded intermediates to accumulate. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.en_US
dc.languageengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/en_US
dc.relation.ispartofJournal of Biological Chemistryen_US
dc.subject.meshBacterial Toxins - Pharmacologyen_US
dc.subject.meshCell Membrane Permeability - Drug Effectsen_US
dc.subject.meshCytosol - Metabolismen_US
dc.subject.meshEndoplasmic Reticulum - Metabolismen_US
dc.subject.meshGlycoproteins - Chemistry - Metabolismen_US
dc.subject.meshHek293 Cellsen_US
dc.subject.meshHemolysin Proteins - Pharmacologyen_US
dc.subject.meshHerpesvirus 4, Human - Enzymologyen_US
dc.subject.meshHumansen_US
dc.subject.meshImmunoblottingen_US
dc.subject.meshProteasome Endopeptidase Complex - Metabolismen_US
dc.subject.meshProtein Foldingen_US
dc.subject.meshProtein Transporten_US
dc.subject.meshSubstrate Specificityen_US
dc.subject.meshUbiquitin - Metabolismen_US
dc.subject.meshViral Proteins - Genetics - Metabolismen_US
dc.titleA viral deubiquitylating enzyme restores dislocation of substrates from the endoplasmic reticulum (ER) in semi-intact cellsen_US
dc.typeArticleen_US
dc.identifier.emailSanyal, S: sumana@wi.mit.eduen_US
dc.identifier.authoritySanyal, S=rp01794en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1074/jbc.M112.365312en_US
dc.identifier.pmid22619172en_US
dc.identifier.scopuseid_2-s2.0-84863620865en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-84863620865&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume287en_US
dc.identifier.issue28en_US
dc.identifier.spage23594en_US
dc.identifier.epage23603en_US
dc.identifier.isiWOS:000306511300030-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridSanyal, S=16069600000en_US
dc.identifier.scopusauthoridClaessen, JHL=24734020000en_US
dc.identifier.scopusauthoridPloegh, HL=35433834100en_US

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