Influence of 3' half-site sequence of NF-kappa B motifs on the binding of lipopolysaccharide-activatable macrophage NF-kappa B proteins.
Lipopolysaccharide treatment of mouse macrophage-like J774 cells was found to result in the activation of three different nuclear proteins which specifically bind to oligonucleotide containing the NF-kappa B motif of the human immunodeficiency virus (HIV) gene. These are designated as NF-kappa B1, -kappa B2, and -kappa B3, according to their electrophoretic mobilities (fast, intermediate, and slow, respectively). Immunological and UV cross-linking studies showed that NF-kappa B1 consists of only p50 subunit, whereas both NF-kappa B2 and -kappa B3 contain NF-kappa B p65 subunit and c-Rel. In addition, NF-kappa B2 was also found to contain p50 subunit of NF-kappa B. The binding of three types of NF-kappa B proteins to HIV NF-kappa B motif was effectively inhibited by other NF-kappa B motifs, whose 3' half-site nucleotide sequences are T/A-T-T/C-CC (HIV, interleukin-6, interferon (INF)-beta, H2-Kb, I-E alpha d, and TNF-alpha 2 (nucleotide position -510)) and much less effectively by NF-kappa B motifs with 3' half-site sequences of TGCCC (TNF-alpha 3, nucleotide position -610), ATCTC (G-CSF), TATTC (Fc gamma R), or TCCTT (TNF-alpha 1, nucleotide position -850). Our data also suggested that NF-kappa B1 and -kappa B2 which contain p50 subunit of NF-kappa B bind with the higher preference for NF-kappa B motif of H2-Kb gene promoter than that of INF-beta, whereas NF-kappa B3 which does not contain p50 subunit appears to preferentially bind to NF-kappa B sites of IFN-beta.
Muroi
M
M
Department of Microbiology, University of Kansas Medical Center, Kansas City 66160-7420.
Muroi
Y
Y
Yamamoto
K
K
Suzuki
T
T
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005260:Female; D015497:HIV-1 / Q000235:genetics; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000302:isolation & purification / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D013379:Substrate Specificity
D008070:Lipopolysaccharides; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides