title: Bleomycin hydrolase is regulated biphasically in a differentiation- and cytokine-dependent manner: relevance to atopic dermatitis.
abstract: Loss-of-function mutation in the profilaggrin gene is a major risk factor for atopic dermatitis (AD). Previously, we showed that a neutral cysteine protease, bleomycin hydrolase (BH), has a role in generating natural moisturizing factors, and calpain I is an upstream protease in the filaggrin degradation pathway. Here, we investigated the transcriptional regulatory mechanisms of BH and the relevance of BH to AD. First, we cloned the 5'-flanking region of BH. Deletion analyses identified a critical region for BH promoter activity within -216 bp upstream. Electrophoretic mobility shift assay revealed that MZF-1, Sp-1, and interferon regulatory factor-1/2 could bind to this region in vitro. Moreover, site-directed mutagenesis of the MZF-1 and Sp-1 motifs markedly reduced BH promoter activity. These data indicate that BH expression is up-regulated via MZF-1 and Sp-1. Interestingly, a Th1 cytokine, IFN-γ, significantly reduced the expression of BH. Analyses with site-directed mutagenesis and small interference RNA supported the suppressing effect of IFN-γ on BH expression. On the other hand, a Th2 cytokine, IL-4, did not show any direct effect on BH expression. However, it down-regulated MZF-1 and Sp-1 in cultured keratinocytes, indicating that IL-4 could work as a suppressor in BH regulation. Lastly, we examined expression of BH in skins of patients with AD. BH activity and expression were markedly decreased in AD lesional skin, suggesting a defect of the filaggrin degradation pathway in AD. Our results suggest that BH transcription would be modulated during both differentiation and inflammation.
authors.affiliation: From the Department of Regulation Biochemistry, Graduate School of Medical Sciences, Kitasato University, Sagamihara, Kanagawa 252-0373, Japan,; the Shiseido Research Center, Yokohama, Kanagawa 238-8643, Japan.
authors.forename: Yayoi
authors.initials: Y
authors.lastname: Kamata
authors.affiliation: the Shiseido Research Center, Yokohama, Kanagawa 238-8643, Japan,; the Department of Dermatology, Tokyo Medical University, Shinjuku, Tokyo 160-8402, Japan.
authors.forename: Mami
authors.initials: M
authors.lastname: Yamamoto
authors.affiliation: From the Department of Regulation Biochemistry, Graduate School of Medical Sciences, Kitasato University, Sagamihara, Kanagawa 252-0373, Japan.
authors.forename: Fumitaka
authors.initials: F
authors.lastname: Kawakami
authors.affiliation: the Department of Dermatology, Tokyo Medical University, Shinjuku, Tokyo 160-8402, Japan.
authors.forename: Ryoji
authors.initials: R
authors.lastname: Tsuboi
authors.affiliation: the Laboratory of Biochemistry, Faculty of Nutritional Sciences, Sagami Women's University, Sagamihara, Kanagawa 252-0383, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Takeda
authors.affiliation: From the Department of Regulation Biochemistry, Graduate School of Medical Sciences, Kitasato University, Sagamihara, Kanagawa 252-0373, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Ishihara
authors.affiliation: the Shiseido Research Center, Yokohama, Kanagawa 238-8643, Japan,. Electronic address: toshihiko.hibino@to.shiseido.co.jp.
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Hibino
mesh_terms: D000998:Antiviral Agents / Q000378:metabolism / Q000494:pharmacology; D001483:Base Sequence; D002454:Cell Differentiation; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics; D003876:Dermatitis, Atopic / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D000091344:Filaggrin Proteins; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism* / Q000494:pharmacology; D015847:Interleukin-4 / Q000378:metabolism* / Q000494:pharmacology; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism; D015603:Keratinocytes / Q000201:enzymology* / Q000473:pathology; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D020218:Response Elements; D017384:Sequence Deletion; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism
chemical_list: D000998:Antiviral Agents; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; C508598:IL4 protein, human; D007381:Intermediate Filament Proteins; D051741:Kruppel-Like Transcription Factors; C085969:MZF1 protein, human; D016329:Sp1 Transcription Factor; D015847:Interleukin-4; D007371:Interferon-gamma; D003546:Cysteine Endopeptidases; C031262:bleomycin hydrolase