Binding of integrin alpha6beta4 to plectin prevents plectin association with F-actin but does not interfere with intermediate filament binding

Geerts, Dirk, Fontao, Lionel, Nievers, Mirjam G., Schaapveld, Roel Q., Purkis, Patricia E., Wheeler, Grant N ORCID: https://orcid.org/0000-0002-4335-8577, Lane, E. Birgitte, Leigh, Irene M. and Sonnenberg, Arnoud (1999) Binding of integrin alpha6beta4 to plectin prevents plectin association with F-actin but does not interfere with intermediate filament binding. Journal of Cell Biology, 147 (2). pp. 417-434. ISSN 0021-9525

Full text not available from this repository. (Request a copy)

Abstract

Hemidesmosomes 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 alpha6beta4 integrin and have shown that the cytoplasmic domain of the beta4 subunit associates with an NH(2)-terminal fragment of plectin that contains the actin-binding domain (ABD). When expressed in immortalized plectin-deficient keratinocytes from human patients with epidermol- ysis bullosa (EB) simplex with muscular dystrophy (MD-EBS), this fragment is colocalized with alpha6beta4 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 beta4 interacts directly with plectin, and identified a major plectin-binding site on the second fibronectin type III repeat of the beta4 cytoplasmic domain. Mapping of the beta4 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 beta4 can compete out the plectin ABD fragment from its association with F-actin. The ability of beta4 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 beta4 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.

Item Type:	Article
Uncontrolled Keywords:	actins,antigens, surface,cell line, transformed,desmosomes,humans,immunohistochemistry,integrin alpha6beta4,integrins,intermediate filament proteins,intermediate filaments,keratinocytes,plectin,protein binding,transfection
Faculty \ School:	Faculty of Science > School of Biological Sciences
UEA Research Groups:	Faculty of Science > Research Groups > Cells and Tissues
Depositing User:	Pure Connector
Date Deposited:	11 Aug 2015 16:10
Last Modified:	19 Apr 2023 00:47
URI:	https://ueaeprints.uea.ac.uk/id/eprint/54065
DOI:	10.1083/jcb.147.2.417

Actions (login required)

View Item