By tethering the intermediate filament (IF) cytoskeleton to the plasma membrane, the desmosome plaque component desmoplakin (DP) strengthens adhesion mediated by the transmembrane desmosomal cadherins. Mutations in DP or its associated armadillo proteins result in potentially lethal disorders of the skin and heart. The loss of mechanical tissue integrity caused by desmosome dysfunction is commonly thought to underlie disease pathogenesis. However, in addition to their mechanical functions, desmosomal molecules provide signaling cues to regulate IF attachment, drive junction assembly, and guide epidermal morphogenesis. The mechanisms by which desmosomes govern signaling pathways to control tissue homeostasis and disease pathogenesis are poorly understood. We propose that the DP N-terminus and associated armadillo proteins in the plakophilin (PKP) family act as scaffolds to harness the activities of signaling mediators when and where they are needed for junction assembly and epidermal differentiation. Our goal is to determine how DP/PKP deficiency and mutations affecting DP-PKP interactions contribute to disease pathogenesis by interfering with structural and signaling functions by: 1) Determining the independent and cooperative roles of DP and PKPs in desmosome assembly and cell- cell adhesion and the effect of human disease mutations that interfere with DP-PKP interactions on these processes, 2) Testing whether DP and PKPs form scaffolds for PKC and small GTPase (RhoA) signaling mediators to integrate effector pathways that control desmosome function and cytoskeletal remodeling, and 3) Elucidating how DP works in conjunction with PKPs to promote epidermal morphogenesis and homeostasis. We will use an shRNA-dependent knock down approach combined with analysis of tissues and cells from mouse models and human patients with DP and PKP deficiencies to establish the respective roles of DP, PKPs and related signaling pathways in differentiation using 2D submerged, 3D in vitro and in vivo transplanted cultures. Desmosome-associated signaling mediators hold promise as targets for the design of small molecule therapies to ameliorate diseases caused by mutations or autoimmune antibodies that co-opt downstream pathways associated with these structural proteins.