Inhibition of ERalpha-mediated trans-activation of human coagulation factor XII gene by heteromeric transcription factor NF-Y.
Human coagulation factor XII promoter contains an estrogen response element that mediates ligand-activated ERalpha induction of coagulation factor XII gene expression. The 3'-half of coagulation factor XII-estrogen response element overlaps a putative CCAAT box, the widespread regulatory element specifically recognized by the heteromeric transcription factor NF-Y. Transient cotransfection of NF-Y and ERalpha results in strong inhibition of estrogen stimulation of coagulation factor XII promoter activity. NF-Y antagonism is primarily exerted by the NF-YA subunit and does not require binding to the CCAAT element, as NF-YA mutants with impaired DNA binding capacity retain the ability to inhibit ERalpha trans-activation. EMSAs with increasing concentrations of recombinant NF-Y do not detect the formation of NF-Y-DNA complexes or show impairment of ERalpha binding to estrogen response element. Immunoprecipitation of whole cell extracts with anti-ERalpha antibody reveals an in vivo association between the two transcription factors, which is abolished by deletion of the NF-YA carboxyl-terminus. In functional experiments with sequential NF-YA deletion mutants the HAP2-homology region appears essential in eliciting NF-YA antagonistic activity. In conclusion, our results demonstrate that heteromeric transcription factor NF-Y inhibits estrogen induction of coagulation factor XII promoter in a DNA binding-independent fashion and suggest a novel role for NF-Y as a partner for the ERalpha transcription complex.
Farsetti
A
A
Istituto di Neurobiologia e Medicina Molecolare, Consiglio Nazionale delle Ricerche, Rome, Italy.
Narducci
M
M
Moretti
F
F
Nanni
S
S
Mantovani
R
R
Sacchi
A
A
Pontecorvi
A
A
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence / Q000235:genetics; D023081:CCAAT-Binding Factor / Q000235:genetics / Q000494:pharmacology / Q000502:physiology*; D004247:DNA / Q000378:metabolism; D004958:Estradiol / Q000494:pharmacology; D047628:Estrogen Receptor alpha; D005174:Factor XII / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000187:drug effects; D020033:Protein Isoforms / Q000235:genetics / Q000494:pharmacology; D017434:Protein Structure, Tertiary / Q000235:genetics; D011960:Receptors, Estrogen / Q000502:physiology*; D014158:Transcription, Genetic / Q000187:drug effects; D015533:Transcriptional Activation / Q000502:physiology*; D014407:Tumor Cells, Cultured
D023081:CCAAT-Binding Factor; D047628:Estrogen Receptor alpha; D020033:Protein Isoforms; D011960:Receptors, Estrogen; D004958:Estradiol; D005174:Factor XII; D004247:DNA