title: Down-regulation of PROS1 gene expression by 17beta-estradiol via estrogen receptor alpha (ERalpha)-Sp1 interaction recruiting receptor-interacting protein 140 and the corepressor-HDAC3 complex.
abstract: Pregnant women show a low level of protein S (PS) in plasma, which is known to be a risk for deep venous thrombosis. 17Beta-estradiol (E(2)), an estrogen that increases in concentration in the late stages of pregnancy, regulates the expression of various genes via the estrogen receptor (ER). Here, we investigated the molecular mechanisms behind the reduction in PS levels caused by E(2) in HepG2-ERalpha cells, which stably express ERalpha, and also the genomic ER signaling pathway, which modulates the ligand-dependent repression of the PSalpha gene (PROS1). We observed that E(2) repressed the production of mRNA and antigen of PS. A luciferase reporter assay revealed that E(2) down-regulated PROS1 promoter activity and that this E(2)-dependent repression disappeared upon the deletion or mutation of two adjacent GC-rich motifs in the promoter. An electrophoretic mobility shift assay and DNA pulldown assay revealed that the GC-rich motifs were associated with Sp1, Sp3, and ERalpha. In a chromatin immunoprecipitation assay, we found ERalpha-Sp protein-promoter interaction involved in the E(2)-dependent repression of PROS1 transcription. Furthermore, we demonstrated that E(2) treatment recruited RIP140 and the NCoR-SMRT-HDAC3 complex to the PROS1 promoter, which hypoacetylated chromatin. Taken together, this suggested that E(2) might repress PROS1 transcription depending upon ERalpha-Sp1 recruiting transcriptional repressors in HepG2-ERalpha cells and, consequently, that high levels of E(2) leading to reduced levels of plasma PS would be a risk for deep venous thrombosis in pregnant women.
authors.affiliation: Department of Pathophysiological Laboratory Sciences, Nagoya University Graduate School of Medicine, Nagoya 461-8673, Japan.
authors.forename: Atsuo
authors.initials: A
authors.lastname: Suzuki
authors.forename: Naomi
authors.initials: N
authors.lastname: Sanda
authors.forename: Yuhri
authors.initials: Y
authors.lastname: Miyawaki
authors.forename: Yuta
authors.initials: Y
authors.lastname: Fujimori
authors.forename: Takayuki
authors.initials: T
authors.lastname: Yamada
authors.forename: Akira
authors.initials: A
authors.lastname: Takagi
authors.forename: Takashi
authors.initials: T
authors.lastname: Murate
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Saito
authors.forename: Tetsuhito
authors.initials: T
authors.lastname: Kojima
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D001798:Blood Proteins / Q000096:biosynthesis* / Q000235:genetics; D004958:Estradiol / Q000494:pharmacology*; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism*; D004967:Estrogens / Q000494:pharmacology*; D005260:Female; D020862:GC Rich Sequence / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects*; D056945:Hep G2 Cells; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D056971:Nuclear Receptor Co-Repressor 1 / Q000235:genetics / Q000378:metabolism; D000078723:Nuclear Receptor Interacting Protein 1; D011247:Pregnancy; D011250:Pregnancy Complications, Hematologic / Q000097:blood / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D017293:Protein S / Q000235:genetics / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D051705:Sp3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D013927:Thrombosis / Q000097:blood / Q000235:genetics; D000099186:Histone Deacetylase 3
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D001798:Blood Proteins; D004958:Estradiol; D047628:Estrogen Receptor alpha; D004967:Estrogens; D006655:Histone Deacetylases; D046912:Multiprotein Complexes; D009687:Nuclear Proteins; D056971:Nuclear Receptor Co-Repressor 1; D000078723:Nuclear Receptor Interacting Protein 1; D017293:Protein S; D016329:Sp1 Transcription Factor; D051705:Sp3 Transcription Factor; D000099186:Histone Deacetylase 3; C539127:NCOR1 protein, human; C000627127:NRIP1 protein, human; C523390:PROS1 protein, human; C497121:SP3 protein, human