NF-kappa B protein purification from bovine spleen: nucleotide stimulation and binding site specificity.
The activity of the enhancer for the kappa immunoglobulin light chain gene critically depends on the presence in the nucleus of the NF-kappa B protein. We purified NF-kappa B over 50,000-fold and identified two protein species, 42 and 44 kDa, that could be eluted and renatured from a sodium dodecyl sulfate/polyacrylamide gel to give specific DNA-binding activity. Binding of the purified bovine NF-kappa B as well as that from human and murine B- or T-lymphoid cell extracts was dramatically stimulated by nucleoside triphosphates. This effect distinguished NF-kappa B from a related factor, H2-TF1. Purified NF-kappa B interacted efficiently with regulatory sequences that function during either B- or T-lymphocyte activation, including the human immunodeficiency virus enhancer and a NF-kappa B binding site we detected in the interleukin 2 enhancer.
Lenardo
M J
MJ
Whitehead Institute for Biomedical Research, Cambridge, MA 02142.
Kuang
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A
Gifford
A
A
Baltimore
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D
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D001483:Base Sequence; D002417:Cattle; D002467:Cell Nucleus / Q000032:analysis; D002845:Chromatography; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism; D017886:Durapatite; D006801:Humans; D006882:Hydroxyapatites; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016328:NF-kappa B; D012265:Ribonucleotides / Q000494:pharmacology; D013154:Spleen / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology; D014157:Transcription Factors / Q000302:isolation & purification* / Q000378:metabolism
D004268:DNA-Binding Proteins; D006882:Hydroxyapatites; D016328:NF-kappa B; D012265:Ribonucleotides; D014157:Transcription Factors; D004247:DNA; D017886:Durapatite