title: Effects of keratin phosphorylation on the mechanical properties of keratin filaments in living cells.
abstract: Keratin filaments impart resilience against mechanical extension of the cell. Despite the pathophysiological relevance of this function, very little is known about the mechanical properties of intermediate filaments in living cells and how these properties are modulated. We used keratin mutants that mimic or abrogate phosphorylation of keratin 8-serine(431) and keratin 18-serine(52) and investigated their effect on keratin tortuousness after cell stretch release in squamous cell carcinoma cells. Cells transfected with the wild-type keratins were used as controls. We can show that keratin dephosphorylation alters the stretch response of keratin in living cells since keratin tortuousness was abolished when phosphorylation of keratin18-serine(52) was abrogated. Additional experiments demonstrate that keratin tortuousness is not simply caused by a plastic overextension of keratin filaments because tortuousness is reversible and requires an intact actin-myosin system. The role of actin in this process remains unclear, but we suggest anchorage of keratin filaments to actin during stretch that leads to buckling on stretch release. Dephosphorylated keratin18-serine(52) might strengthen the recoil force of keratin filaments and hence explain the abolished buckling. The almost exclusive immunolabeling for phosphorylated keratin18-serine (52) in the cell periphery points at a particular role of the peripheral keratin network in this regard.
authors.affiliation: Institute for General Physiology, Ulm University, Ulm, Germany.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Fois
authors.forename: Michael
authors.initials: M
authors.lastname: Weimer
authors.forename: Tobias
authors.initials: T
authors.lastname: Busch
authors.forename: Erika T
authors.initials: ET
authors.lastname: Felder
authors.forename: Franz
authors.initials: F
authors.lastname: Oswald
authors.forename: Götz
authors.initials: G
authors.lastname: von Wichert
authors.forename: Thomas
authors.initials: T
authors.lastname: Seufferlein
authors.forename: Paul
authors.initials: P
authors.lastname: Dietl
authors.forename: Edward
authors.initials: E
authors.lastname: Felder
mesh_terms: D000199:Actins / Q000378:metabolism*; D002470:Cell Survival; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism*; D007633:Keratins / Q000378:metabolism* / Q000648:ultrastructure; D010766:Phosphorylation / Q000502:physiology; D012694:Serine / Q000378:metabolism
chemical_list: D000199:Actins; D012694:Serine; D007633:Keratins