Please use this identifier to cite or link to this item: https://doi.org/10.1083/jcb.147.2.417
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dc.titleBinding of integrin α6β4 to plectin prevents plectin association with f-actin but does not interfere with intermediate filament binding
dc.contributor.authorGeerts, D
dc.contributor.authorFontao, L
dc.contributor.authorNievers, M.G
dc.contributor.authorSchaapveld, R.Q.J
dc.contributor.authorPurkis, P.E
dc.contributor.authorWheeler, G.N
dc.contributor.authorLane, E.B
dc.contributor.authorLeigh, I.M
dc.contributor.authorSonnenberg, A
dc.date.accessioned2020-10-27T09:58:24Z
dc.date.available2020-10-27T09:58:24Z
dc.date.issued1999
dc.identifier.citationGeerts, D, Fontao, L, Nievers, M.G, Schaapveld, R.Q.J, Purkis, P.E, Wheeler, G.N, Lane, E.B, Leigh, I.M, Sonnenberg, A (1999). Binding of integrin α6β4 to plectin prevents plectin association with f-actin but does not interfere with intermediate filament binding. Journal of Cell Biology 147 (2) : 417-434. ScholarBank@NUS Repository. https://doi.org/10.1083/jcb.147.2.417
dc.identifier.issn00219525
dc.identifier.urihttps://scholarbank.nus.edu.sg/handle/10635/181141
dc.description.abstractHemidesmosomes are stable adhesion complexes in basal epithelial cells that provide a link between the intermediate filament network and the extracellular matrix. We have investigated the recruitment of plectin into hemidesmosomes by the α6β4 integrin and have shown that the cytoplasmic domain of the β4 subunit associates with an NH2-terminal fragment of plectin that contains the actin-binding domain (ABD). When expressed in immortalized plectin-deficient keratinocytes from human patients with epidermolysis bullosa (EB) simplex with muscular dystrophy (MD-EBS), this fragment is colocalized with α6β4 in basal hemidesmosome-like clusters or associated with F-actin in stress fibers or focal contacts. We used a yeast two-hybrid binding assay in combination with an in vitro dot blot overlay assay to demonstrate that β4 interacts directly with plectin, and identified a major plectin-binding site on the second fibronectin type III repeat of the β4 cytoplasmic domain. Mapping of the β4 and actin-binding sites on plectin showed that the binding sites overlap and are both located in the plectin ABD. Using an in vitro competition assay, we could show that β4 can compete out the plectin ABD fragment from its association with F-actin. The ability of β4 to prevent binding of F-actin to plectin explains why F-actin has never been found in association with hemidesmosomes, and provides a molecular mechanism for a switch in plectin localization from actin filaments to basal intermediate filament-anchoring hemidesmosomes when β4 is expressed. Finally, by mapping of the COOH-terminally located binding site for several different intermediate filament proteins on plectin using yeast two-hybrid assays and cell transfection experiments with MD-EBS keratinocytes, we confirm that plectin interacts with different cytoskeletal networks.
dc.rightsAttribution 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.sourceUnpaywall 20201031
dc.subjectactin
dc.subjectf actin
dc.subjectfibronectin
dc.subjectintegrin
dc.subjectplectin
dc.subjectprotein subunit
dc.subjectactin
dc.subjectalpha6beta4 integrin
dc.subjectintegrin
dc.subjectintermediate filament protein
dc.subjectmembrane antigen
dc.subjectPLEC1 protein, human
dc.subjectplectin
dc.subjectamino terminal sequence
dc.subjectarticle
dc.subjectbinding site
dc.subjectcarboxy terminal sequence
dc.subjectcell immortalization
dc.subjectcontrolled study
dc.subjectcytoplasm
dc.subjectcytoskeleton
dc.subjectepidermolysis bullosa simplex
dc.subjecthemidesmosome
dc.subjecthuman
dc.subjecthuman cell
dc.subjectintermediate filament
dc.subjectkeratinocyte
dc.subjectmuscular dystrophy
dc.subjectnonhuman
dc.subjectnucleotide sequence
dc.subjectpriority journal
dc.subjectprotein binding
dc.subjectprotein domain
dc.subjectprotein localization
dc.subjectyeast
dc.subjectcell line
dc.subjectdesmosome
dc.subjectgenetic transfection
dc.subjectimmunohistochemistry
dc.subjectkeratinocyte
dc.subjectmetabolism
dc.subjectprotein binding
dc.subjectultrastructure
dc.subjectActins
dc.subjectAntigens, Surface
dc.subjectCell Line, Transformed
dc.subjectDesmosomes
dc.subjectHumans
dc.subjectImmunohistochemistry
dc.subjectIntegrin alpha6beta4
dc.subjectIntegrins
dc.subjectIntermediate Filament Proteins
dc.subjectIntermediate Filaments
dc.subjectKeratinocytes
dc.subjectPlectin
dc.subjectProtein Binding
dc.subjectTransfection
dc.typeArticle
dc.contributor.departmentPATHOLOGY
dc.description.doi10.1083/jcb.147.2.417
dc.description.sourcetitleJournal of Cell Biology
dc.description.volume147
dc.description.issue2
dc.description.page417-434
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