Activation of the transcription factor NF-KB in GH3 pituitary cells.
Since several genes expressed in the pituitary can bind the transcription factor NF-KB, its presence and regulation was examined in the GH3 pituitary cell line. An electrophoretic mobility shift assay using nuclear extracts and an oligonucleotide probe corresponding to the Ig KB binding site was employed to identify activated NF-KB. One complex possessed properties characteristic of NF-KB: co-migration with an NF-KB complex and binding specificity restricted to NF-KB binding DNA sequences. Antibodies to the NF-KB subunits NFKB1p50 (p50) and RelA (p65) interacted with the extract-DNA complex. Activation of NF-KB in GH3 cells was increased by PMA or the cytokine tumor necrosis factor alpha. A synergy between PMA and TNF or a calcium mobilizing agent was seen in NF-KB activation. Further TNF activation was enhanced by TRH. These observations indicate the presence of NF-KB in GH3 cells and demonstrate its activation by hormones/second messengers that act on pituitary cells.
Grandison
L
L
Department of Physiology and Biophysics, Robert Wood Johnson Medical School-UMDNJ, Piscataway 08854-5635.
Nolan
G P
GP
Pfaff
D W
DW
D001483:Base Sequence; D001665:Binding Sites; D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D002467:Cell Nucleus / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism*; D015345:Oligonucleotide Probes; D010902:Pituitary Gland / Q000378:metabolism* / Q000648:ultrastructure; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D016328:NF-kappa B; D015345:Oligonucleotide Probes; D014409:Tumor Necrosis Factor-alpha; D000001:Calcimycin; D013973:Thyrotropin-Releasing Hormone; D004247:DNA; D013755:Tetradecanoylphorbol Acetate