title: Keratin 8 modulation of desmoplakin deposition at desmosomes in hepatocytes.
abstract: Keratins, the intermediate filament proteins of epithelial cells, connect to desmosomes, the cell-cell adhesion structures at the surface membrane. The building elements of desmosomes include desmoglein and desmocollin, which provide the actual cell adhesive properties, and desmoplakins, which anchor the keratin intermediate filaments to desmosomes. In the work reported here, we address the role of keratin 8 in modulating desmoplakin deposition at surface membrane in mouse hepatocytes. The experimental approach is based on the use of keratin 8- and keratin 18-null mouse hepatocytes as cell models. In wild-type mouse hepatocytes, desmoplakin is aligned with desmoglein and keratin 8 at the surface membrane. In keratin 8-null hepatocytes, the intermediate filament loss leads to alterations in desmoplakin distribution at the surface membrane, but not of desmoglein. Intriguingly, a significant proportion of keratin 18-null hepatocytes express keratin 8 at the surface membrane, associated with a proper desmoplakin alignment with desmoglein at desmosomes. A Triton treatment of the monolayer reveals that most of the desmoplakin present in either wild-type, keratin 8- or keratin 18-null hepatocytes is insoluble. Deletion analysis of keratin 8 further suggests that the recovery of desmoplakin alignment requires the keratin 8 rod domain. In addition, similarly to other works revealing a key role of desmoplakin phosphorylation on its interaction with intermediate filaments, we find that the phosphorylation status of the keratin 8 head domain affects desmoplakin distribution at desmosomes. Together, the data indicate that a proper alignment/deposition of desmoplakin with keratins and desmoglein in hepatocytes requires keratin 8, through a reciprocal phosphoserine-dependent process.
authors.affiliation: Centre de recherche en cancérologie, QC, Canada G1R 2J6.
authors.forename: Anne
authors.initials: A
authors.lastname: Loranger
authors.forename: Stéphane
authors.initials: S
authors.lastname: Gilbert
authors.forename: Jean-Simon
authors.initials: JS
authors.lastname: Brouard
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Magin
authors.forename: Normand
authors.initials: N
authors.lastname: Marceau
mesh_terms: D000818:Animals; D002462:Cell Membrane; D002478:Cells, Cultured; D051180:Desmoplakins / Q000378:metabolism*; D003896:Desmosomes / Q000378:metabolism*; D022781:Hepatocytes / Q000378:metabolism*; D053538:Keratin-18 / Q000502:physiology*; D053533:Keratin-8 / Q000235:genetics / Q000502:physiology*; D051379:Mice; D010766:Phosphorylation; D017434:Protein Structure, Tertiary
chemical_list: D051180:Desmoplakins; D053538:Keratin-18; D053533:Keratin-8