Identification of DNA-binding proteins that interact with the 5'-flanking region of the human D-amino acid oxidase gene by pull-down assay coupled with two-dimensional gel electrophoresis and mass spectrometry.
D-Amino acid oxidase (DAO) is a flavoenzyme that metabolizes D-amino acids and is expected to be a promising therapeutic target of schizophrenia and glioblastoma. The study of DNA-binding proteins has yielded much information in the regulation of transcription and other biological processes. However, proteins interacting with DAO gene have not been elucidated. Our assessment of human DAO promoter activity using luciferase reporter system indicated the 5'-flanking region of this gene (-4289 bp from transcription initiation site) has a regulatory sequence for gene expression, which is regulated by multi-protein complexes interacting with this region. By using pull-down assay coupled with two-dimensional gel electrophoresis and mass spectrometry, we identified six proteins binding to the 5'-flanking region of the human DAO gene (zinc finger C2HC domain-containing protein 1A; histidine-tRNA ligase, cytoplasmic; molybdenum cofactor biosynthesis protein; 60S ribosomal protein L37; calponin-1; calmodulin binding protein and heterogeneous nuclear ribonucleoprotein A2/B1). These preliminary results will contribute to the advance in the understanding of the potential factors associated with the regulatory mechanism of DAO expression.
Division of Enzyme Pathophysiology, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
Diem Hong
DH
Tran
Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
Yuji
Y
Shishido
Division of Enzyme Pathophysiology, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
Seong Pil
SP
Chung
Division of Enzyme Pathophysiology, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
Huong Thi Thanh
HT
Trinh
Division of Enzyme Pathophysiology, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
Kazuko
K
Yorita
Division of Enzyme Pathophysiology, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima, 3-18-15 Kuramoto, Tokushima 770-8503, Japan.
Takashi
T
Sakai
Division of Enzyme Pathophysiology, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima, 3-18-15 Kuramoto, Tokushima 770-8503, Japan; Division of Enzyme Literacy, The Institute for Enzyme Research (KOSOKEN), The University of Tokushima, 3-18-15 Kuramoto, Tokushima 770-8503, Japan; Japan Science and Technology Agency, CREST, 3-18-15 Kuramoto, Tokushima 770-8503, Japan. Electronic address: kiyo.fukui@tokushima-u.ac.jp.
Kiyoshi
K
Fukui
D000818:Animals; D003605:D-Amino-Acid Oxidase / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000032:analysis* / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D018374:LLC-PK1 Cells; D013058:Mass Spectrometry / Q000379:methods; D011485:Protein Binding / Q000502:physiology; D013552:Swine
D004268:DNA-Binding Proteins; D003605:D-Amino-Acid Oxidase
5′-Flanking region; DNA-binding protein; Mass spectrometry; Pull-down assay; d-Amino acid oxidase