Involvement of NF-Y in transcriptional regulation of the phospholamban gene.
To understand the transcriptional regulation of the phospholamban gene, we analyzed a 5'-upstream region of the gene. Using a series of deletion constructs, we demonstrated that the region from -96 bp to -78 bp, containing the CCAAT sequence, is essential for transcription of this gene. This region specifically bound to nuclear proteins extracted from rat hearts, and gel-shift assays using competitive oligonucleotides, antibodies and recombinant proteins showed that this region binds to the NF-YA and NF-YB, members of the CCAAT-binding nuclear protein family. This region-dependent transcription in cardiac myocytes transfected with antisense cDNAs encoding NF-YA and NF-YB was decreased to approximately 50% of that seen in cells transfected with the same sense cDNAs. We, therefore, conclude that the region from -96 bp to -78 bp plays a critical role in expression of the phospholamban gene, which is regulated by binding of the nuclear protein NF-Y.
Yabuki
M
M
Department of Medicine and Pathophysiology, Osaka University Medical School, Suita, Japan.
Toyofuku
T
T
Otsu
K
K
Nishida
M
M
Kuzuya
T
T
Hori
M
M
Tada
M
M
D000818:Animals; D001483:Base Sequence; D022762:CCAAT-Enhancer-Binding Proteins; D002135:Calcium-Binding Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D009687:Nuclear Proteins / Q000032:analysis; D016376:Oligonucleotides, Antisense; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics; D000099211:Phospholamban
D022762:CCAAT-Enhancer-Binding Proteins; D002135:Calcium-Binding Proteins; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D016376:Oligonucleotides, Antisense; D011994:Recombinant Proteins; D014157:Transcription Factors; D000099211:Phospholamban