title: Formation of hemidesmosome-like structures in the absence of ligand binding by the (alpha)6(beta)4 integrin requires binding of HD1/plectin to the cytoplasmic domain of the (beta)4 integrin subunit.
abstract: Hemidesmosomes are adhesion structures that mediate anchorage of epithelial cells to the underlying basement membrane. We have previously shown that the (alpha)6(beta)4 integrin can induce the assembly of these multi-protein structures independent of binding to its ligand laminin-5 (ligand-independent formation of hemidesmosomes). Our results suggested a role for HD1/plectin, which binds to the cytoplasmic domain of the (beta)4 integrin subunit, in controlling the clustering of hemidesmosomal components at the basal side of the cell. Using keratinocytes derived from patients lacking HD1/plectin, we now show that ligand-independent formation of hemidesmosomal clusters indeed requires HD1/plectin, in contrast to the ligand-dependent assembly of hemidesmosomes. No clustering of the (alpha)6(beta)4 integrin, or of the bullous pemphigoid antigens BP180 and BP230, was seen when HD1/plectin-deficient keratinocytes were plated on fibronectin or type IV collagen. In (&bgr;)4-deficient keratinocytes, expression of an interleukin 2 receptor (IL2R) transmembrane chimera containing the (beta)4 cytoplasmic tail with the mutation R1281W, which abrogates HD1/plectin binding, resulted in a diffuse distribution of the chimeric receptor. In contrast, a (beta)4(R1281W) mutant that can associate with (alpha)6 and bind ligand, was found to be directed to the basal surface of the cells, at sites where laminin-5 was deposited. In addition, this mutant induced clustering of BP180 and BP230 at these sites. Together, these results show that the formation of hemidesmosomes requires binding of either ligand or HD1/plectin to the (beta)4 integrin subunit. Intriguingly, we found that IL2R/(beta)4 chimeras become localized in pre-existing hemidesmosomes of HD1/plectin-deficient keratinocytes, and that this localization requires a domain in the (beta)4 cytoplasmic tail that is also required for HD1/plectin binding (residues 1115-1356). Because this part of (beta)4 lacks the BP180 binding site, and since we show in this study that it is unable to interact with the same part on another (beta)4 molecule, we suggest that the chimera becomes incorporated into hemidesmosomes of HD1/plectin-deficient keratinocytes by interacting with an as yet unidentified hemidesmosomal component.
authors.affiliation: The Netherlands Cancer Institute, Division of Cell Biology, Plesmanlaan 121, The Netherlands.
authors.forename: M G
authors.initials: MG
authors.lastname: Nievers
authors.forename: I
authors.initials: I
authors.lastname: Kuikman
authors.forename: D
authors.initials: D
authors.lastname: Geerts
authors.forename: I M
authors.initials: IM
authors.lastname: Leigh
authors.forename: A
authors.initials: A
authors.lastname: Sonnenberg
mesh_terms: D015703:Antigens, CD / Q000502:physiology; D000954:Antigens, Surface / Q000502:physiology*; D002461:Cell Line, Transformed; D003896:Desmosomes / Q000502:physiology* / Q000648:ultrastructure*; D006801:Humans; D039161:Integrin alpha6beta4; D039663:Integrin beta4; D016023:Integrins / Q000502:physiology*; D007381:Intermediate Filament Proteins / Q000502:physiology*; D015603:Keratinocytes / Q000502:physiology / Q000648:ultrastructure*; D008024:Ligands; D051190:Plectin
chemical_list: D015703:Antigens, CD; D000954:Antigens, Surface; D039161:Integrin alpha6beta4; D039663:Integrin beta4; D016023:Integrins; D007381:Intermediate Filament Proteins; D008024:Ligands; C495332:PLEC protein, human; D051190:Plectin