Estrogen-occupied estrogen receptor represses cyclin G2 gene expression and recruits a repressor complex at the cyclin G2 promoter.
Estrogens, acting through their nuclear receptors have a broad impact on target cells, eliciting a transcriptional response program that involves gene repression as well as gene stimulation. While much is known about the mechanisms by which the estrogen-occupied estrogen receptor (ER) stimulates gene expression, the molecular events that lead to gene repression by the hormone-ER complex are largely unknown. Because estradiol represses expression of the cyclin G2 gene, which encodes a negative regulator of the cell cycle, our aim was to understand the mechanism by which cyclin G2 is repressed by estrogen. We show that cyclin G2 is a primary ER target gene in MCF-7 breast cancer cells that is rapidly and robustly down-regulated by estrogen. Promoter analysis reveals a responsive region containing a half-estrogen response element and GC-rich region that interact with ER and Sp1 proteins. Mutation of the half-ERE abrogates hormone-mediated repression. Mutational mapping of receptor reveals a requirement for its N-terminal region and DNA binding domain to support cyclin G2 repression. Following estradiol treatment of cells, chromatin immunoprecipitation analyses reveal recruitment of ER to the cyclin G2 regulatory region, dismissal of RNA polymerase II, and recruitment of a complex containing N-CoR and histone deacetylases, leading to a hypoacetylated chromatin state. Our study provides evidence for a mechanism by which the estrogen-occupied ER is able to actively repress gene expression in vivo and indicates a role for nuclear receptor corepressors and associated histone deacetylase activity in mediating negative gene regulation by this hormone-occupied nuclear receptor.
Stossi
Fabio
F
Department of Molecular and Integrative Physiology, Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Likhite
Varsha S
VS
Katzenellenbogen
John A
JA
Katzenellenbogen
Benita S
BS
D045744:Cell Line, Tumor; D056767:Cyclin G2; D016213:Cyclins / Q000235:genetics / Q000378:metabolism*; D004967:Estrogens / Q000378:metabolism*; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D008954:Models, Biological; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D011960:Receptors, Estrogen / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000378:metabolism
C101593:CCNG2 protein, human; D056767:Cyclin G2; D016213:Cyclins; D004967:Estrogens; D011960:Receptors, Estrogen; D016329:Sp1 Transcription Factor; D012319:RNA Polymerase II; D006655:Histone Deacetylases