title: Crucial roles of MZF1 and Sp1 in the transcriptional regulation of the peptidylarginine deiminase type I gene (PADI1) in human keratinocytes.
abstract: Peptidylarginine deiminases (PADs) catalyze the conversion of protein-bound arginine residues into citrulline residues in a calcium-dependent manner. The PAD1 gene (PADI1) is expressed in a few tissues, including the epidermis, where the protein is detected with a higher level in the more differentiated keratinocytes. Using quantitative reverse transcription-PCR experiments, we show that PADI1 mRNAs are more abundant in keratinocytes cultured with 1.2 than 0.15 mM calcium. We cloned and characterized the promoter region using human keratinocytes transfected with variously deleted fragments of the 5'-upstream region of PADI1 coupled to the luciferase gene. We found that as few as 195 bp upstream from the transcription initiation site were sufficient to direct transcription of the reporter gene. Mutations of MZF1- or Sp1-binding sites markedly reduced PADI1 promoter activity. Chromatin immunoprecipitation assays revealed that MZF1 and Sp1/Sp3 bind to this region in vivo. Furthermore, MZF1 or Sp1 small interfering RNAs (siRNAs) effectively diminished PADI1 expression in keratinocytes cultured in both low- and high-calcium-containing medium. In addition, the expression of MZF1 and PAD1 increased in parallel when normal human epidermal keratinocytes underwent differentiation. These data indicate that MZF1 and Sp1/Sp3 binding to the promoter region drive the PADI1 expression.
authors.affiliation: Department of Applied Biological Resource Sciences, School of Agriculture, Ibaraki University, Ami-machi, Inashiki-gun, Ibaraki, Japan.
authors.forename: Sijun
authors.initials: S
authors.lastname: Dong
authors.forename: Shibo
authors.initials: S
authors.lastname: Ying
authors.forename: Toshio
authors.initials: T
authors.lastname: Kojima
authors.forename: Masakazu
authors.initials: M
authors.lastname: Shiraiwa
authors.forename: Akira
authors.initials: A
authors.lastname: Kawada
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: Méchin
authors.forename: Véronique
authors.initials: V
authors.lastname: Adoue
authors.forename: Stéphane
authors.initials: S
authors.lastname: Chavanas
authors.forename: Guy
authors.initials: G
authors.lastname: Serre
authors.forename: Michel
authors.initials: M
authors.lastname: Simon
authors.forename: Hidenari
authors.initials: H
authors.lastname: Takahara
mesh_terms: D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D000078404:Epidermal Cells; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D006867:Hydrolases / Q000235:genetics*; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology*; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000502:physiology; D000080062:Protein-Arginine Deiminase Type 1; D034741:RNA, Small Interfering; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation / Q000502:physiology
chemical_list: D051741:Kruppel-Like Transcription Factors; C085969:MZF1 protein, human; D034741:RNA, Small Interfering; D016329:Sp1 Transcription Factor; D006867:Hydrolases; D000080062:Protein-Arginine Deiminase Type 1; D002118:Calcium