GATA-3 represses gp91phox gene expression in eosinophil-committed HL-60-C15 cells.
To study the regulatory mechanism of gp91phox gene expression in eosinophils, we transiently transfected eosinophil-committed HL-60-C15 cells with gp91phox promoter constructs, and identified a negative element from bp -267 to -246 of the gp91phox gene, the deletion of which caused an 83% increase in promoter activity. Electrophoresis mobility shift assays demonstrated GATA-3 binds to the GATA consensus site from bp -256 to -250. An 81% increment in promoter activity was obtained when a mutation was introduced in the GATA-3 binding site of the bp -267 to +12 construct, which is comparable to that of the bp -245 to +12 construct. We therefore conclude that GATA-3 specifically binding to the GATA site negatively regulates the expression of the gene in HL-60-C15 cells.
Sadat
M A
MA
Department of Pediatrics, Nagasaki University School of Medicine, Sakamoto, Japan. f1106@cc.nagasaki-u.ac.jp
Kumatori
A
A
Suzuki
S
S
Yamaguchi
Y
Y
Tsuji
Y
Y
Nakamura
M
M
D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004804:Eosinophils / Q000166:cytology / Q000378:metabolism*; D050990:GATA3 Transcription Factor; D005786:Gene Expression Regulation; D018922:HL-60 Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000235:genetics; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015534:Trans-Activators / Q000378:metabolism*; D014162:Transfection; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; D050990:GATA3 Transcription Factor; C494749:GATA3 protein, human; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases