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Papers In Press, published online ahead of print April 4, 2006
Center for Craniofacial Molecular Biology, University of Southern California, Los Angeles, CA 90033
Corresponding Author: mlsnead{at}hsc.usc.edu
Amelogenin is the major protein component of the forming enamel matrix. In situ hybridization revealed a periodicity for amelogenin mRNA hybridization signals, ranging from low to high transcript abundance on serial sections of developing mouse teeth. This in vivo observation led us to examine the amelogenin promoter for the activity of transcription factor(s) that account for this expression aspect of the regulation for amelogenin gene. We have previously shown that CCAAT/enhancer-binding protein alpha (C/EBPa) is a potent transactivator of the mouse X-chromosomal amelogenin gene acting at the C/EBPa cis-element located in the –70/+52 minimal promoter. The minimal promoter contains a reversed CCAAT box (-58/-54), which is four base pairs downstream from the C/EBPa binding site. Similar to the C/EBPa binding site, the integrity of the reversed CCAAT box is also required for maintaining the activity of the basal promoter. We therefore focused on transcription factors that interact with the reversed CCAAT box. Using electrophoretic mobility shift assays we demonstrated that NF-Y was directly bound to this reversed CCAAT site. Co-transfection of C/EBPa and NF-Y synergistically increased the promoter activity. In contrast, increased expression of NF-Y alone had only marginal effects on the promoter. A dominant-negative DNA binding-deficient NF-Y mutant (NF-YAm29) dramatically decreased the promoter activity, both in the absence or presence of exogenous expression of C/EBPa. We identified protein-protein interactions between C/EBPa with NF-Y by a co-immunoprecipitation analysis. These results suggest that C/EBPa and NF-Y synergistically activate the mouse amelogenin gene and can contribute to its physiological regulation during amelogenesis
J. Biol. Chem, 10.1074/jbc.M510514200
Submitted on September 26, 2005
Accepted on March 31, 2006
NF-Y and CCAAT/enhancer-binding protein
synergistically activate the mouse amelogenin gene
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