title: Breaking the connection: caspase 6 disconnects intermediate filament-binding domain of periplakin from its actin-binding N-terminal region.
abstract: Periplakin is a member of the plakin family of cytolinkers that connect cytoskeletal networks to each other as well as to the cell junctional complexes. Here, we demonstrate a direct molecular interaction between actin and periplakin. Furthermore, the oligomerization state of periplakin was shown to determine specificity of its binding to intermediate filaments (IF) in vitro. Both the filament association and the cell membrane localization of periplakin were confirmed in the cells overexpressing human periplakin. Double labeling of the N- and C-terminally tagged periplakin revealed unexpected lack of co-localization of periplakin ends in a confluent culture, and separation of the periplakin ends was even more pronounced in apoptotic cells. Western analysis revealed that after induction of apoptosis, periplakin becomes cleaved close to its C-terminal tail. Only the distinct cleavage products, but not the full-length periplakin, were present in the cells detached from the solid support during the apoptotic process. We show that caspase 6 cleaves periplakin at an unconventional recognition site, amino acid sequence TVAD. Thus, the separation of periplakin ends disconnects the actin-binding head-rod domain from the IF-binding C-terminal domain. We show that specific cleavage products co-exist with the full-length periplakin in cells, suggesting physiological consequences due to their altered binding specificities.
authors.affiliation: Laboratory of Skin Biology, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Andrey E
authors.initials: AE
authors.lastname: Kalinin
authors.forename: Alexandr E
authors.initials: AE
authors.lastname: Kalinin
authors.forename: Mikko
authors.initials: M
authors.lastname: Aho
authors.forename: Jouni
authors.initials: J
authors.lastname: Uitto
authors.forename: Sirpa
authors.initials: S
authors.lastname: Aho
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000502:physiology; D053178:Caspase 6; D020169:Caspases / Q000378:metabolism*; D003598:Cytoskeletal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004285:Dogs; D015870:Gene Expression; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism*; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D051179:Plakins; D017434:Protein Structure, Tertiary
chemical_list: D000199:Actins; D003598:Cytoskeletal Proteins; D051179:Plakins; C110037:Ppl protein, mouse; C505514:CASP6 protein, human; C505516:Casp6 protein, mouse; D053178:Caspase 6; D020169:Caspases