title: Multiple pathways are involved in DNA degradation during keratinocyte terminal differentiation.
abstract: Loss of the nucleus is a critical step in keratinocyte terminal differentiation. To elucidate the mechanisms involved, we focused on two characteristic events: nuclear translocation of N-terminal fragment of profilaggrin and caspase-14-dependent degradation of the inhibitor of caspase-activated DNase (ICAD). First, we demonstrated that epidermal mesotrypsin liberated a 55-kDa N-terminal fragment of profilaggrin (FLG-N) and FLG-N was translocated into the nucleus. Interestingly, these cells became TUNEL positive. Mutation in the mesotrypsin-susceptible Arg-rich region between FLG-N and the first filaggrin domain abolished these changes. Furthermore, caspase-14 caused limited proteolysis of ICAD, followed by accumulation of caspase-activated DNase (CAD) in TUNEL-positive nuclei. Knockdown of both proteases resulted in a significant increase of remnant nuclei in a skin equivalent model. Immunohistochemical study revealed that both caspase-14 and mesotrypsin were markedly downregulated in parakeratotic areas of lesional skin from patients with atopic dermatitis and psoriasis. Collectively, our results indicate that at least two pathways are involved in the DNA degradation process during keratinocyte terminal differentiation.
authors.affiliation: 1] Shiseido Research Center, 2-2-1 Hayabuchi, Tsuzuki-ku, Yokohama 224-8558, Japan [2] Department of Dermatology, Tokyo Medical University, 6-7-1 Nishishinjuku, Shinjuku-ku, Tokyo 160-0023, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Yamamoto-Tanaka
authors.affiliation: Department of Dermatology, University of Toyama, Toyama 930-0194, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Makino
authors.affiliation: Shiseido Research Center, 2-2-1 Hayabuchi, Tsuzuki-ku, Yokohama 224-8558, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Motoyama
authors.affiliation: Shiseido Research Center, 2-2-1 Hayabuchi, Tsuzuki-ku, Yokohama 224-8558, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Miyai
authors.affiliation: Department of Dermatology, Tokyo Medical University, 6-7-1 Nishishinjuku, Shinjuku-ku, Tokyo 160-0023, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Tsuboi
authors.affiliation: Shiseido Research Center, 2-2-1 Hayabuchi, Tsuzuki-ku, Yokohama 224-8558, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Hibino
mesh_terms: D053456:Caspase 14 / Q000378:metabolism; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D004247:DNA / Q000378:metabolism; D053938:DNA Fragmentation; D015536:Down-Regulation; D004817:Epidermis / Q000378:metabolism / Q000473:pathology; D000091344:Filaggrin Proteins; D055785:Gene Knockdown Techniques; D006801:Humans; D020287:In Situ Nick-End Labeling; D007381:Intermediate Filament Proteins / Q000737:chemistry / Q000378:metabolism; D015603:Keratinocytes / Q000378:metabolism* / Q000473:pathology; D008954:Models, Biological; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D020449:Repetitive Sequences, Amino Acid; D015398:Signal Transduction; D014162:Transfection; D014362:Trypsinogen / Q000378:metabolism
chemical_list: C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D007381:Intermediate Filament Proteins; D014362:Trypsinogen; D004247:DNA; D053456:Caspase 14
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