JBC Transcription and Nuclear Factor Monoclonals

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J Biol Chem, Vol. 274, Issue 7, 4147-4154, February 12, 1999

A Short Conserved Motif Is Required for Repressor Domain Function in the Myeloid-specific Transcription Factor CCAAT/Enhancer-binding Protein epsilon

Nicholas D. Angerer, Yang Du, Demet Nalbant, and Simon C. WilliamsDagger

From the Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center and Dagger  Southwest Cancer Center at University Medical Center, Lubbock, Texas 79430

CCAAT/enhancer-binding protein epsilon  (C/EBPepsilon ) is expressed almost exclusively in the myeloid lineage of the hematopoietic system and functions during terminal differentiation of neutrophils and macrophages, and in the regulation of cytokine gene expression in macrophages and T cells. We have undertaken a series of structure/function studies on the murine C/EBPepsilon polypeptide to investigate the mechanism by which C/EBPepsilon activates transcription. Studies with deletion mutants and fusion proteins consisting of C/EBPepsilon sequences joined to the Gal4 DNA-binding protein identified two transcriptional activation domains in C/EBPepsilon . Removal of sequences between the two activation domains or sequences between the second activation domain and the C-terminal DNA binding domain significantly increased the activity of C/EBPepsilon , suggesting the presence of two separate regulatory domains (designated RD-1epsilon and RD-2epsilon ). RD-1epsilon behaved as a classic active repressor domain being capable of inhibiting adjacent activation domains irrespective of their origin and when linked to a heterologous DNA binding domain. Mutagenesis studies revealed a short motif in RD-1epsilon that appears to be a target site for protein-protein interactions and is conserved in repressor domains from C/EBPbeta , Sp3, c-Fos, and FosB. The juxtaposition of activation and repressor domains may permit C/EBPepsilon to function as a transcriptional activator or repressor at different stages of myeloid differentiation or as an inducible transcriptional activator of cytokine genes.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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