Identification of proteins within the nuclear factor-kappa B transcriptional complex including estrogen receptor-alpha
American Journal of Obstetrics and Gynecology
NLM Title Abbreviation
Am J Obstet Gynecol
OBJECTIVE: The objective of the study was to determine whether cross-talk occurs between estrogen receptors (ERs) and nuclear factor-kappa-B (NF-kappaB), to assess the functional consequences of such an ER/NF-kappaB interaction, and to identify other unknown regulatory proteins that may participate in the NF-kappaB transcriptional complex. STUDY DESIGN: Electromobility gel shifts, reporter gene assays, and mass spectrometry were used to identify proteins interacting with the NF-kappaB deoxyribonucleic acid (DNA) response element. RESULTS: ER and the p65 subunit of NF-kappaB colocalized on DNA. This interaction was inhibitory for ER transcriptional activity. Sequencing of proteins bound to the NF-kappaB/DNA complex identified DNA-modifying enzymes, scaffolding proteins, chaperones, and elements of the nuclear matrix. CONCLUSION: These studies have identified an inhibitory interaction between estrogen receptors and the p65 subunit of NF-kappaB with implications for estrogen action in pregnancy and cancer. New accessory proteins have also been identified that bind to protein complexes on the NF-kappaB DNA response element.
Cells, Cultured, Electrophoretic Mobility Shift Assay, Endometrial Neoplasms/genetics/metabolism, Estrogen Receptor alpha/genetics/metabolism, Female, Gene Expression Regulation, Genes, Reporter, Humans, NF-kappa B/genetics/metabolism, Polymerase Chain Reaction, Pregnancy, Probability, Protein Binding, Sampling Studies, Sensitivity and Specificity, Signal Transduction, Transcriptional Activation, Trophoblasts/cytology
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