title: Identification of two promoters for human D-amino acid oxidase gene: implication for the differential promoter regulation mediated by PAX5/PAX2.
abstract: D-amino acid oxidase (DAO) is a flavoenzyme that metabolizes d-amino acids. Until now, the DAO expression mechanism is still unclear. Our assessment of human DAO (hDAO) promoter activity using luciferase reporter system indicated the proximal upstream region of exon1 (-237/+1) has promoter activity (P1). Interestingly, we identified an alternative promoter in the proximal upstream region of exon2 (+4,126/+4,929) (P2). This alternative promoter has stronger activity than that of P1. Our results also revealed a negative regulatory segment (+1,163/+1,940) in intron1; that would act in concert with P1 and P2. Bioinformatics analyses elucidated the conservation of transcription factor PAX5 family binding sites among species. These sites (-60/-31) and (+4,464/+4,493), locate in P1 and P2 of hDAO, respectively. Gel shift assays demonstrated P1 contains a site (-60/-31) for PAX5 binding while P2 has three sites for both paired box gene 2 (PAX2) and paired box gene 5 (PAX5) binding. The dual roles of PAX5 family in regulating hDAO transcription by modulating promoter activity of P1 and activating promoter activity of P2 were implicated based on the site-directed mutagenesis experiment. Altogether, our data suggested the differential regulation of hDAO expression by two promoters whose activities may be modulated by the binding of PAX2 and PAX5.
authors.affiliation: Division of Enzyme Pathophysiology; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima; and Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
authors.forename: Diem Hong
authors.initials: DH
authors.lastname: Tran
authors.affiliation: Division of Enzyme Pathophysiology; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima; and Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Shishido
authors.affiliation: Division of Enzyme Pathophysiology; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima; and Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
authors.forename: Seong Pil
authors.initials: SP
authors.lastname: Chung
authors.affiliation: Division of Enzyme Pathophysiology; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima; and Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
authors.forename: Huong Thi Thanh
authors.initials: HT
authors.lastname: Trinh
authors.affiliation: Division of Enzyme Pathophysiology; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima; and Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
authors.forename: Kazuko
authors.initials: K
authors.lastname: Yorita
authors.affiliation: Division of Enzyme Pathophysiology; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima; and Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Sakai
authors.affiliation: Division of Enzyme Pathophysiology; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima; and Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan Division of Enzyme Pathophysiology; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima; and Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan Division of Enzyme Pathophysiology; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima; and Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Fukui
mesh_terms: D000818:Animals; D001483:Base Sequence; D003605:D-Amino-Acid Oxidase / Q000235:genetics*; D006801:Humans; D018374:LLC-PK1 Cells; D008969:Molecular Sequence Data; D051762:PAX2 Transcription Factor / Q000235:genetics*; D051757:PAX5 Transcription Factor / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012689:Sequence Homology, Nucleic Acid; D013552:Swine
chemical_list: D051762:PAX2 Transcription Factor; C497318:PAX2 protein, human; D051757:PAX5 Transcription Factor; C497281:PAX5 protein, human; D003605:D-Amino-Acid Oxidase
keywords: PAX2; PAX5; d-amino acid oxidase; promoter; transcription factor