Identification of transcriptional activation and repression domains in human CCAAT/enhancer-binding protein ε

Elizabeth A. Williamson, Haixin N. Xu, Adrian F. Gombart, Walter Verbeek, Alexey M. Chumakov, Alan David Friedman, H. Phillip Koeffler

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)14796-14804
Number of pages9
JournalJournal of Biological Chemistry
Volume273
Issue number24
DOIs
StatePublished - Jun 12 1998

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CCAAT-Enhancer-Binding Proteins
Transcriptional Activation
Chemical activation
Transcription
Genes
Chloramphenicol O-Acetyltransferase
Reporter Genes
Amino Acids
Bearings (structural)
Fusion reactions
Aminoacylation
Leukocyte Elastase
Luciferases
Yeast
Peroxidase
Up-Regulation
Yeasts
Binding Sites
Cells
Cell Line

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of transcriptional activation and repression domains in human CCAAT/enhancer-binding protein ε. / Williamson, Elizabeth A.; Xu, Haixin N.; Gombart, Adrian F.; Verbeek, Walter; Chumakov, Alexey M.; Friedman, Alan David; Koeffler, H. Phillip.

In: Journal of Biological Chemistry, Vol. 273, No. 24, 12.06.1998, p. 14796-14804.

Research output: Contribution to journalArticle

Williamson, Elizabeth A. ; Xu, Haixin N. ; Gombart, Adrian F. ; Verbeek, Walter ; Chumakov, Alexey M. ; Friedman, Alan David ; Koeffler, H. Phillip. / Identification of transcriptional activation and repression domains in human CCAAT/enhancer-binding protein ε. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 24. pp. 14796-14804.
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