title: Transcriptional activation of rat creatine kinase B by 17beta-estradiol in MCF-7 cells involves an estrogen responsive element and GC-rich sites.
abstract: The rat creatine kinase B (CKB) gene is induced by estrogen in the uterus, and constructs containing rat CKB gene promoter inserts are highly estrogen-responsive in cell culture. Analysis of the upstream -568 to -523 region of the promoter in HeLa cells has identified an imperfect palindromic estrogen response element (ERE) that is required for hormone inducibility. Analysis of the CKB gene promoter in MCF-7 breast cancer cells confirmed that pCKB7 (containing the -568 to -523 promoter insert) was estrogen-responsive in transient transfection studies. However, mutation and deletion analysis of this region of the promoter showed that two GC-rich sites and the concensus ERE were functional cis-elements that bound estrogen receptor alpha (ERalpha)/Sp1 and ERalpha proteins, respectively. The role of these elements was confirmed in gel mobility shift and chromatin immunoprecipitation assays and transfection studies in MDA-MB-231 and Schneider Drosophila SL-2 cells. These results show that transcriptional activation of CKB by estrogen is dependent, in part, on ERalpha/Sp1 action which is cell context-dependent.
authors.affiliation: Department of Veterinary Physiology & Pharmacology, Texas A&M University, College Station, TX 77843-4466, USA.
authors.forename: F
authors.initials: F
authors.lastname: Wang
authors.forename: I
authors.initials: I
authors.lastname: Samudio
authors.forename: S
authors.initials: S
authors.lastname: Safe
mesh_terms: D000818:Animals; D001483:Base Sequence; D001943:Breast Neoplasms; D002460:Cell Line; D003402:Creatine Kinase / Q000235:genetics*; D052276:Creatine Kinase, BB Form; D004252:DNA Mutational Analysis; D004330:Drosophila; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D004958:Estradiol / Q000494:pharmacology*; D047628:Estrogen Receptor alpha; D005260:Female; D020862:GC Rich Sequence; D006801:Humans; D007527:Isoenzymes; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011960:Receptors, Estrogen / Q000235:genetics; D017384:Sequence Deletion; D016329:Sp1 Transcription Factor / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects*; D014407:Tumor Cells, Cultured
chemical_list: D047628:Estrogen Receptor alpha; D007527:Isoenzymes; D011960:Receptors, Estrogen; D016329:Sp1 Transcription Factor; D004958:Estradiol; D003402:Creatine Kinase; D052276:Creatine Kinase, BB Form