Identification of transcription factor in the promoter region of rat regucalcin gene: binding of nuclear factor I-A1 to TTGGC motif.
Hepatic nuclear protein has been reported to bind specifically to the TTGGC sequence of the rat regucalcin gene promoter region in stimulating the promoter activity (Misawa and Yamaguchi [2000] Biochem. Biophys. Res. Commun. 279: 275-281). The present study was undertaken to identify transcription factor, which binds to TTGGC motif in the rat regucalcin gene promoter, using the yeast one-hybrid system. The sequence between -525 and -504, which has been defined as a functional promoter element II-b, was used as bait to screen a rat liver cDNA library. Two cDNA clones were identified as a nuclear factor I-A1 (NF1-A1). The results of gel mobility shift assay and mutation analysis using recombinant NF1-A1 protein showed that this protein could specifically bind to TTGGC motif of the II-b oligonucleotide in promoter region. The expression of NF1-A1 mRNA was found in the liver, kidney, heart, spleen, and brain of rats. This study demonstrates that NF1-A1 is a transcription factor in stimulating the rat regucalcin gene promoter activity.
Misawa
Hiroyuki
H
Laboratory of Endocrinology and Molecular Metabolism, Graduate School of Nutritional Sciences, University of Shizuoka, 52-1 Yada, Shizuoka 422-8526, Japan.
Yamaguchi
Masayoshi
M
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D022762:CCAAT-Enhancer-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000235:genetics*; D002265:Carboxylic Ester Hydrolases; D006528:Carcinoma, Hepatocellular; D004268:DNA-Binding Proteins; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D047908:Intracellular Signaling Peptides and Proteins; D008969:Molecular Sequence Data; D051839:NFI Transcription Factors; D009841:Oligonucleotides / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics; D015238:Sulfotransferases; D014018:Tissue Distribution / Q000502:physiology; D014157:Transcription Factors / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D020798:Two-Hybrid System Techniques / Q000639:trends; D051840:Y-Box-Binding Protein 1
D022762:CCAAT-Enhancer-Binding Proteins; D002135:Calcium-Binding Proteins; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D051839:NFI Transcription Factors; D009841:Oligonucleotides; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D051840:Y-Box-Binding Protein 1; C486169:Ybx1 protein, rat; D015238:Sulfotransferases; C028152:alcohol sulfotransferase; D002265:Carboxylic Ester Hydrolases; C513325:Rgn protein, rat