TY - JOUR
T1 - The adult enhancer factor-1, a Drosophila melanogaster transcriptional repressor, modulates the promoter activity of the rat class-I alcohol dehydrogenase-encoding gene
AU - Potter, James J.
AU - Mezey, Esteban
AU - Yang, Vincent W.
N1 - Funding Information:
We thank Drs. Ted Abel, Tom Maniatis, M. Daniel Lane and StevenL . McKnight for reagentsT. his work was in part supportedb y U.S. Public Health Service grantsA A00626( to E.M.) and AA09334( to V.W.Y.), and a grant-in-aid from the Alcoholic Beverage Medical ResearchF oundation( to V.W.Y.).
Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 1994/11/18
Y1 - 1994/11/18
N2 - Expression of the Drosophila melanogaster alcohol dehydrogenase-encoding gene (ADH) in the adult fat body is controlled by the ADH adult enhancer site (AAE). The D. melanogaster transcription repressor, adult enhancer factor-1 (AEF-1), binds to AAE at a site which overlaps with a sequence recognized by the mammalian transcription factor, CCAAT/enhancer-binding protein α [C/EBPα; Falb and Maniatis, Genes Dev. 6 (1992a) 454-465]. C/EBPα also activates the promoter of the rat class-I ADH gene in a sequence-specific manner [Potter et al., Arch. Biochem. Biophys. 285 (1991a) 246-251]. In this study, we explored the possibility that D. melanogaster AEF-1 influences transcription of the rat class-I ADH. By DNase I footprint analysis, bacterially produced AEF-1 protects a region of DNA between nucleotides (nt) -22 and - 36 of the rat class-I ADH promoter (pADH), just 5′ to the binding site of C/EBPα, a result confirmed by the electrophoretic mobility shift assay (EMSA). Co-transfection of a rat pADH-CAT reporter construct with expression vectors containing C/EBPα, AEF-1, or both, indicates that AEF-1 inhibits induction of the rat pADH by C/EBPa. Moreover, rat liver nuclear extracts appear to contain AEF-1-like-binding activities to AAE by EMSA. These experiments suggest an evolutionarily conserved mechanism by which AEF-1 modulates expression of the D. melanogaster and rat ADH genes.
AB - Expression of the Drosophila melanogaster alcohol dehydrogenase-encoding gene (ADH) in the adult fat body is controlled by the ADH adult enhancer site (AAE). The D. melanogaster transcription repressor, adult enhancer factor-1 (AEF-1), binds to AAE at a site which overlaps with a sequence recognized by the mammalian transcription factor, CCAAT/enhancer-binding protein α [C/EBPα; Falb and Maniatis, Genes Dev. 6 (1992a) 454-465]. C/EBPα also activates the promoter of the rat class-I ADH gene in a sequence-specific manner [Potter et al., Arch. Biochem. Biophys. 285 (1991a) 246-251]. In this study, we explored the possibility that D. melanogaster AEF-1 influences transcription of the rat class-I ADH. By DNase I footprint analysis, bacterially produced AEF-1 protects a region of DNA between nucleotides (nt) -22 and - 36 of the rat class-I ADH promoter (pADH), just 5′ to the binding site of C/EBPα, a result confirmed by the electrophoretic mobility shift assay (EMSA). Co-transfection of a rat pADH-CAT reporter construct with expression vectors containing C/EBPα, AEF-1, or both, indicates that AEF-1 inhibits induction of the rat pADH by C/EBPa. Moreover, rat liver nuclear extracts appear to contain AEF-1-like-binding activities to AAE by EMSA. These experiments suggest an evolutionarily conserved mechanism by which AEF-1 modulates expression of the D. melanogaster and rat ADH genes.
KW - CCAAT/enhancer-binding protein
KW - DNase I footprint
KW - cat reporter
KW - competition
KW - electrophoretic mobility shift assay
KW - repression
KW - transcription factor
KW - transfection
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U2 - 10.1016/0378-1119(94)90170-8
DO - 10.1016/0378-1119(94)90170-8
M3 - Article
C2 - 7959011
AN - SCOPUS:0028043497
SN - 0378-1119
VL - 149
SP - 325
EP - 330
JO - Gene
JF - Gene
IS - 2
ER -