The tetraspan molecule CD151, a novel constituent of hemidesmosomes, associates with the integrin α6β4 and may regulate the spatial organization of hemidesmosomes

CD151 is a cell surface protein that belongs to the tetraspan superfamily. It associates with other tetraspan molecules and certain integrins to form large complexes at the cell surface. CD151 is expressed by a variety of epithelia and mesenchymal cells. We demonstrate here that in human skin CD151 is codistributed with alpha 3 beta 1 and alpha 6 beta 4 at the basolateral surface of basal keratinocytes, Immunoelectron microscopy showed that CD151 is concentrated in hemidesmosomes. By immunoprecipitation from transfected K562 cells, we established that CD151 associates with alpha 3 beta 1 and alpha 6 beta 4. In beta 4-deficient pyloric atresia associated with junctional epidermolysis bullosa (PA-JEB) keratinocytes, CD151 and alpha 3 beta 1 are clustered together at the basal cell surface in association with patches of laminin-5. Focal adhesions are present at the periphery of these clusters, connected with actin filaments, and they contain both CD151 and alpha 3 beta 1. Transient transfection studies of PA-JEB cells with beta 4 revealed that the integrin alpha 6 beta 4 becomes incorporated into the alpha 3 beta 1-CD151 clusters where it induces the formation of hemidesmosomes. As a result, the amount of alpha 3 beta 1 in the clusters diminishes and the protein becomes restricted to the peripheral focal adhesions. Furthermore, CD151 becomes predominantly associated with alpha 6 beta 4 in hemidesmosomes, whereas its codistribution with alpha 3 beta 1 in focal adhesions becomes partial. The localization of alpha 6 beta 4 in the pre-hemidesmosomal clusters is accompanied by a strong upregulation of CD151, which is at least partly due to increased cell surface expression. Using beta 4 chimeras containing the extracellular and transmembrane domain of the IL-2 receptor and the cytoplasmic domain of beta 4, we found that for recruitment of CD151 into hemidesmosomes, the beta 4 subunit must be associated with alpha 6, confirming that integrins associate with tetraspans via their or subunits. CD151 is the only tetraspan identified in hemidesmosomal structures. Others, such as CD9 and CD81, remain diffusely distributed at the cell surface. In conclusion, we show that CD151 is a major component of (pre)-hemidesmosomal structures and that its recruitment into hemidesmosomes is regulated by the integrin alpha 6 beta 4. We suggest that CD151 plays a role in the formation and stability of hemidesmosomes by providing a framework for the spatial organization of the different hemidesmosomal components.