Nuclear factor Y activates the human xanthine oxidoreductase gene promoter.
To study the regulation of the human xanthine oxidoreductase (XOR) gene, we cloned 1.9 kb of the promoter region. In reporter gene assays, a construct encompassing nucleotides between 142 to +42 conferred maximal basal activity of the XOR promoter in 293T cells, in comparison with shorter (-92 to +42) or longer (up to -1937 to +42) constructs. The promoter activity was low in NIH-3T3 cells. The most active construct contained a putative CCAAT motif at -119 to -123. Electrophoretic mobility shift assays showed that this sequence binds the ubiquitous nuclear factor Y (NF-Y). Mutation of the CCAAT motif (CTGAT) abolished the NF-Y binding and considerably reduced the promoter activity. Our data suggest an important functional role for NF-Y in the transcriptional activation of the human XOR gene.
Martelin
E
E
Hospital for Children and Adolescents, Research Laboratory, University of Helsinki, Finland. eeva.martelin@hus.fi
Palvimo
J J
JJ
Lapatto
R
R
Raivio
K O
KO
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D023081:CCAAT-Binding Factor / Q000378:metabolism*; D002461:Cell Line, Transformed; D004247:DNA / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D051381:Rats; D014157:Transcription Factors / Q000378:metabolism*; D014968:Xanthine Dehydrogenase / Q000235:genetics*; D014969:Xanthine Oxidase / Q000235:genetics*
D023081:CCAAT-Binding Factor; D014157:Transcription Factors; C402244:nuclear factor Y; D004247:DNA; D014968:Xanthine Dehydrogenase; D014969:Xanthine Oxidase