The WNT7b promoter is regulated by TTF-1, GATA6, and Foxa2 in lung epithelium.
In this study, we find that WNT7b is the only member of the WNT family of autocrine/paracrine signaling molecules whose expression in the lung is restricted to the airway epithelium during embryonic development. To study the transcriptional mechanisms that underlie this restricted pattern of WNT7b expression, we isolated the proximal 1.0-kb mouse WNT7b promoter and mapped the transcriptional start sites. Transfection of the lung epithelial cell line MLE-15, which expresses WNT7b, shows that the 1.0-kb mouse WNT7b promoter is highly active in lung epithelial cells. This region of the WNT7b promoter contains several DNA binding sites for the important lung-restricted transcription factors TTF-1, GATA6, and Foxa2. Electrophoretic mobility shift assays showed that TTF-1, GATA6, and Foxa2 can bind to a specific subset of their consensus DNA binding sites within the WNT7b promoter. Using cotransfection assays, we demonstrate that TTF-1, GATA6, and Foxa2 can trans-activate the WNT7b promoter in NIH-3T3 cells. Truncation of GATA6 or Foxa2 binding sites reduced the ability of these transcriptional regulators to trans-activate the WNT7b promoter. Finally, the minimal 118-bp region of the mouse WNT7b promoter containing only TTF-1 binding sites was synergistically activated by TTF-1 and GATA6, and we show that TTF-1 and GATA6 physically interact in vivo. Together, these results suggest that WNT7b gene expression in the lung epithelium is regulated in a combinatorial fashion by TTF-1, GATA6, and Foxa2.
Weidenfeld
Joel
J
Department of Medicine, Molecular Cardiology Research Center, and the Department of Dermatology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Shu
Weiguo
W
Zhang
Lili
L
Millar
Sarah E
SE
Morrisey
Edward E
EE
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004247:DNA; D004268:DNA-Binding Proteins / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay; D004848:Epithelium / Q000378:metabolism; D050993:GATA6 Transcription Factor; D005786:Gene Expression Regulation / Q000502:physiology*; D006023:Glycoproteins; D051542:Hepatocyte Nuclear Factor 3-beta; D008168:Lung / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000502:physiology*; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000737:chemistry / Q000235:genetics*; D000074482:Thyroid Nuclear Factor 1; D014157:Transcription Factors / Q000502:physiology*; D051153:Wnt Proteins
D004268:DNA-Binding Proteins; C496503:Foxa2 protein, mouse; D050993:GATA6 Transcription Factor; C494760:Gata6 protein, mouse; D006023:Glycoproteins; C000614414:Nkx2-1 protein, mouse; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D000074482:Thyroid Nuclear Factor 1; D014157:Transcription Factors; D051153:Wnt Proteins; C095937:Wnt7b protein, mouse; D051542:Hepatocyte Nuclear Factor 3-beta; D004247:DNA