TY - JOUR
T1 - Estrogen receptor-KRAB chimeras are potent ligand-dependent repressors of estrogen-regulated gene expression
AU - De Haan, Georgius
AU - Chusacultanachai, Sudsanguan
AU - Mao, Chengjian
AU - Katzenellenbogen, Benita S.
AU - Shapiro, David J.
PY - 2000/5/5
Y1 - 2000/5/5
N2 - As an approach to targeted repression of genes of interest, we describe the development of human estrogen receptor (ER) α-KRAB repressor domain chimeras that are potent ligand-dependent repressors of the transcription of estrogen response element (ERE)-containing promoters and analyze their mechanisms of action. Repression by the KRAB domain was dominant over transactivation mediated by ER AF1 and AF2. An ERE and an ER ligand (estrogen or antiestrogen) were required for repression. Studies with several promoters and cell lines demonstrated that the presence of EREs, rather than the capacity for estrogen induction, determines the potential for repression of a gene by the KRAB-ERα-KRAB (KERK) chimera. A single consensus ERE was sufficient for repression, but the KERK chimera was unable to suppress transcription from the imperfect ERE in the native pS2 promoter. We recently reported mutations that enhance binding of asteroid receptor DNA-binding domain to the ERE. Introducing these mutations into wild-type ER enhanced transactivation from the pS2 ERE. Insertion of these mutations into KERK created the novel repressor KERK-3M, which is a potent repressor of both ER- induced and basal transcription on a promoter containing the pS2 ERE. These modified ER-KRAB chimeras should prove useful as new tools for the functional analysis and repression of ER-regulated genes.
AB - As an approach to targeted repression of genes of interest, we describe the development of human estrogen receptor (ER) α-KRAB repressor domain chimeras that are potent ligand-dependent repressors of the transcription of estrogen response element (ERE)-containing promoters and analyze their mechanisms of action. Repression by the KRAB domain was dominant over transactivation mediated by ER AF1 and AF2. An ERE and an ER ligand (estrogen or antiestrogen) were required for repression. Studies with several promoters and cell lines demonstrated that the presence of EREs, rather than the capacity for estrogen induction, determines the potential for repression of a gene by the KRAB-ERα-KRAB (KERK) chimera. A single consensus ERE was sufficient for repression, but the KERK chimera was unable to suppress transcription from the imperfect ERE in the native pS2 promoter. We recently reported mutations that enhance binding of asteroid receptor DNA-binding domain to the ERE. Introducing these mutations into wild-type ER enhanced transactivation from the pS2 ERE. Insertion of these mutations into KERK created the novel repressor KERK-3M, which is a potent repressor of both ER- induced and basal transcription on a promoter containing the pS2 ERE. These modified ER-KRAB chimeras should prove useful as new tools for the functional analysis and repression of ER-regulated genes.
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U2 - 10.1074/jbc.275.18.13493
DO - 10.1074/jbc.275.18.13493
M3 - Article
C2 - 10788463
AN - SCOPUS:0034607833
SN - 0021-9258
VL - 275
SP - 13493
EP - 13501
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 18
ER -