Characterization of the human zinc finger protein 267 promoter: essential role of nuclear factor Y.
Liver fibrosis results from an excessive deposition of extracellular matrix proteins secreted by activated hepatic stellate cells (HSCs). The activation process is accompanied by an increased activity of various transcription factors, including zinc finger protein 267 (ZNF267). Recently, ZNF267 has been shown to modulate gene expression and to function as a transcriptional repressor. MMP-10 was identified as a target gene; its gene expression and promoter activity are inhibited by ZNF267, which might promote liver fibrogenesis through diminished matrix degradation. However, the transcriptional regulation of the ZNF267 gene is unknown. In the present study, we have cloned and characterized the human ZNF267 promoter containing a 1.5 kb fragment of the 5'-flanking region (-1414/+173). The ZNF267 gene has a TATA-less promoter with multiple transcription initiation sites. Analysis of serial 5'-deletions of luciferase reporter constructs revealed a minimal promoter between -72 and +173 bp. Mutational analysis of putative regulatory elements indicated that a CCAAT box within this region was essential for ZNF267 promoter activity. Electrophoretic mobility shift assays demonstrated that transcription factor nuclear factor Y (NF-Y) bound to the CCAAT box. In co-transfection experiments, NF-YA increased the promoter activity of ZNF267. In conclusion, our results suggest that the binding site for NF-Y is critical for ZNF267 gene regulation and, herewith, the activation of this transcriptional factor may play an important role in the activation process of HSCs and in liver fibrosis.
Hu
Kanghong
K
Department of Internal Medicine I, University of Regensburg, 93042 Regensburg, Germany.
Fink
Marina
M
Froh
Matthias
M
Gäbele
Erwin
E
Hellerbrand
Claus
C
Mühlbauer
Marcus
M
Wiest
Reiner
R
Schölmerich
Jürgen
J
Schnabl
Bernd
B
D001665:Binding Sites / Q000235:genetics / Q000502:physiology; D023081:CCAAT-Binding Factor / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D006801:Humans; D008099:Liver / Q000166:cytology / Q000502:physiology*; D008103:Liver Cirrhosis / Q000235:genetics / Q000378:metabolism; D053506:Matrix Metalloproteinase 10; D008666:Metalloendopeptidases / Q000096:biosynthesis; D012097:Repressor Proteins / Q000235:genetics*; D020218:Response Elements; D017384:Sequence Deletion; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics
D023081:CCAAT-Binding Factor; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; C502002:ZNF267 protein, human; C402244:nuclear factor Y; D008666:Metalloendopeptidases; D053506:Matrix Metalloproteinase 10