title: A role for disulfide bonding in keratin intermediate filament organization and dynamics in skin keratinocytes.
abstract: We recently reported that a trans-dimer, homotypic disulfide bond involving Cys367 in keratin 14 (K14) occurs in an atomic-resolution structure of the interacting K5/K14 2B domains and in keratinocyte cell lines. Here we show that a sizable fraction of the K14 and K5 protein pools participates in interkeratin disulfide bonding in primary cultures of mouse skin keratinocytes. By comparing the properties of wild-type K14 with a completely cysteine-free variant thereof, we found that K14-dependent disulfide bonding limited filament elongation during polymerization in vitro but was necessary for the genesis of a perinuclear-concentrated network of keratin filaments, normal keratin cycling, and the sessile behavior of the nucleus and whole cell in keratinocytes studied by live imaging. Many of these phenotypes were rescued when analyzing a K14 variant harboring a single Cys residue at position 367. These findings establish disulfide bonding as a novel and important mechanism regulating the assembly, intracellular organization, and dynamics of K14-containing intermediate filaments in skin keratinocytes.
authors: Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health; and Department of Biological Chemistry and Department of Dermatology, School of Medicine, Johns Hopkins University, Baltimore, MD 21205.
authors: Xia
authors: X
authors: Feng
authors: Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health; and Department of Biological Chemistry and Department of Dermatology, School of Medicine, Johns Hopkins University, Baltimore, MD 21205 Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health; and Department of Biological Chemistry and Department of Dermatology, School of Medicine, Johns Hopkins University, Baltimore, MD 21205 Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health; and Department of Biological Chemistry and Department of Dermatology, School of Medicine, Johns Hopkins University, Baltimore, MD 21205 coulombe@jhu.edu.
authors: Pierre A
authors: PA
authors: Coulombe
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D003553:Cystine / Q000502:physiology*; D007382:Intermediate Filaments / Q000378:metabolism / Q000648:ultrastructure*; D053547:Keratin-14 / Q000378:metabolism / Q000648:ultrastructure*; D015603:Keratinocytes / Q000378:metabolism / Q000648:ultrastructure*; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D012867:Skin / Q000166:cytology
chemical_list: D053547:Keratin-14; D003553:Cystine