TY - JOUR
T1 - Identification of transcriptional activation and repression domains in human CCAAT/enhancer-binding protein ε
AU - Williamson, Elizabeth A.
AU - Xu, Haixin N.
AU - Gombart, Adrian F.
AU - Verbeek, Walter
AU - Chumakov, Alexey M.
AU - Friedman, Alan D.
AU - Koeffler, H. Phillip
N1 - Copyright:
Copyright 2007 Elsevier B.V., All rights reserved.
PY - 1998/6/12
Y1 - 1998/6/12
N2 - Human CCAAT/enhancer-binding protein ε (C/EBPε), a new member of the C/EBP family, significantly upregulates both the mim-1 and human myeloperoxidase promoters, suggesting an important role for C/EBPε in the transcriptional regulation of a subset of myeloid-specific genes. To elucidate the structure and function of C/EBPε in transcriptional activation, amino acid residues 1-115, 147-249, or 1-249 of C/EBPε were fused to the yeast GAL4 DNA binding domain. These expression vectors were cotransfected with a chloramphenicol acetyltransferase reporter gene and, in all cell lines tested, only the GAL-C/EBPε-(1-115) fusion protein significantly activated expression from the chloramphenicol acetyltransferase reporter gene. Sixteen deletion mutants of C/EBPε mapped the transactivation domain to amino acids 1-18 at the N terminus and revealed the presence of a transcription repression element between amino acid residues 116 and 162. Expression vectors containing the repression domain of C/EBPε strongly inhibited gene transcription from TK, SV40, and adenovital major late promoters bearing GALA binding sites. Fusion of this repression domain to the VP16 activation domain inhibited the transactivation function of VP16. Deletion of this repression domain increased gene transcription from a neutrophil elastase promoter-luciferase reporter. Taken together, these data suggest that C/EBPε regulates transcription by utilizing both activation and repression functions.
AB - Human CCAAT/enhancer-binding protein ε (C/EBPε), a new member of the C/EBP family, significantly upregulates both the mim-1 and human myeloperoxidase promoters, suggesting an important role for C/EBPε in the transcriptional regulation of a subset of myeloid-specific genes. To elucidate the structure and function of C/EBPε in transcriptional activation, amino acid residues 1-115, 147-249, or 1-249 of C/EBPε were fused to the yeast GAL4 DNA binding domain. These expression vectors were cotransfected with a chloramphenicol acetyltransferase reporter gene and, in all cell lines tested, only the GAL-C/EBPε-(1-115) fusion protein significantly activated expression from the chloramphenicol acetyltransferase reporter gene. Sixteen deletion mutants of C/EBPε mapped the transactivation domain to amino acids 1-18 at the N terminus and revealed the presence of a transcription repression element between amino acid residues 116 and 162. Expression vectors containing the repression domain of C/EBPε strongly inhibited gene transcription from TK, SV40, and adenovital major late promoters bearing GALA binding sites. Fusion of this repression domain to the VP16 activation domain inhibited the transactivation function of VP16. Deletion of this repression domain increased gene transcription from a neutrophil elastase promoter-luciferase reporter. Taken together, these data suggest that C/EBPε regulates transcription by utilizing both activation and repression functions.
UR - http://www.scopus.com/inward/record.url?scp=0032511054&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0032511054&partnerID=8YFLogxK
U2 - 10.1074/jbc.273.24.14796
DO - 10.1074/jbc.273.24.14796
M3 - Article
C2 - 9614080
AN - SCOPUS:0032511054
SN - 0021-9258
VL - 273
SP - 14796
EP - 14804
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 24
ER -