title: Unique role for the periplakin tail in intermediate filament association: specific binding to keratin 8 and vimentin.
abstract: Plectin, desmoplakin, and the 230-kDa bullous pemphigoid antigen (BPAG1), members of the plakin family of proteins, are multifunctional cytolinkers, connecting the cytoskeletal structures to the cell adhesion complexes. Envoplakin and periplakin are components of the cornified envelope, but less is known about their role in tissues other than the stratified epithelium. Our tissue-wide survey utilizing RT-PCR revealed that periplakin, like plectin and desmoplakin, has a wide tissue distribution, but envoplakin expression is limited to certain tissues only, and BPAG1 is clearly specific for epidermal keratinocytes. Plectin, desmoplakin and BPAG1 are known to bind to the intermediate filaments through their C-terminal domains. The short C-terminal domain of periplakin is composed only of the linker domain, a region highly homologous between the plakin proteins. Here we demonstrate, through the use of yeast two-hybrid assay, a specific interaction of the periplakin linker domain with keratin 8 and vimentin. Co-expression of each plakin linker domain with keratin 8 revealed that periplakin and BPAG1 linkers co-localize with keratin signals in HaCaT cells, plectin and desmoplakin linkers were detected both in the nucleus and in cytoplasm together with the overexpressed keratin 8, while envoplakin linker localized independently into the nucleus. These results suggest that, in spite of its high homology and structural similarity with envoplakin, periplakin is functionally closer to the well-characterized plakin proteins plectin and desmoplakin, and thus may function tissue-wide as a scaffolding protein in intermediate filament assembly.
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism*; D053533:Keratin-8; D015603:Keratinocytes / Q000378:metabolism*; D007633:Keratins / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051179:Plakins; D011485:Protein Binding; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014018:Tissue Distribution; D014162:Transfection; D020798:Two-Hybrid System Techniques; D014746:Vimentin / Q000378:metabolism*
chemical_list: D003598:Cytoskeletal Proteins; D018076:DNA, Complementary; C426203:KRT8 protein, human; D053533:Keratin-8; C486595:Krt8 protein, mouse; D008565:Membrane Proteins; C491674:PPL protein, human; D051179:Plakins; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D014746:Vimentin; C100754:envoplakin; D007633:Keratins