Autosomal recessive mutations in the cytolinker protein plectin account for the multisystem disorders epidermolysis bullosa simplex (EBS) associated with muscular dystrophy (EBS-MD), pyloric atresia (EBS-PA), and congenital myasthenia (EBS-CMS). In contrast, a dominant missense mutation leads to the disease EBS-Ogna, manifesting exclusively as skin fragility. We have exploited this trait to study the molecular basis of hemidesmosome failure in EBS-Ogna and to reveal the contribution of plectin to hemidesmosome homeostasis. We generated EBS-Ogna knock-in mice mimicking the human phenotype and show that blistering reflects insufficient protein levels of the hemidesmosome-associated plectin isoform 1a. We found that plectin 1a, in contrast to plectin 1c, the major isoform expressed in epidermal keratinocytes, is proteolytically degraded, supporting the notion that degradation of hemidesmosome-anchored plectin is spatially controlled. Using recombinant proteins, we show that the mutation renders plectin's 190-nm-long coiled-coil rod domain more vulnerable to cleavage by calpains and other proteases activated in the epidermis but not in skeletal muscle. Accordingly, treatment of cultured EBS-Ogna keratinocytes as well as of EBS-Ogna mouse skin with calpain inhibitors resulted in increased plectin 1a protein expression levels. Moreover, we report that plectin's rod domain forms dimeric structures that can further associate laterally into remarkably stable (paracrystalline) polymers. We propose focal self-association of plectin molecules as a novel mechanism contributing to hemidesmosome homeostasis and stabilization.
Hemidesmosomes are specialized protein complexes that promote anchorage of the basal keratinocyte cell layer of the epidermis to the underlying dermis. They provide tissue integrity and resistance to mechanical forces. When hemidesmosomes do not function properly, skin blistering ensues in response to mechanical trauma. Plectin is an essential component of hemidesmosomes. Humans carrying recessive mutations in the plectin gene most frequently develop multisystem disorders, where in addition to skin other tissues are also affected. However, there is a unique dominant plectin mutation, which leads to the disease epidermolysis bullosa simplex Ogna (EBS-Ogna), affecting skin exclusively. Because of that, EBS-Ogna is an exceptional system to study the contribution of plectin to hemidesmosome function. We have generated an EBS-Ogna mouse model that mimics the human disease. Using this model, we have learned that selective degradation of hemidesmosome-associated plectin isoform 1a by proteases activated specifically in keratinocytes results in reduced numbers and dysfunction of hemidesmosomes. In contrast, plectin-1c, another plectin isoform expressed in keratinocytes, is not degraded. Moreover, we find that plectin dimers can oligomerize via their long coiled-coil rod domain, a process likely to be instrumental in maintenance of hemidesmosome integrity. These findings highlight the importance of plectin-1a for hemidesmosome function.