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 f ound to result in the activation of three different nuclear proteins w hich specifically bind to oligonucleotide containing the NF-kappaB mot if of the human immunodeficiency virus (HIV) gene. These are designate d as NF-kappaB1, -kappaB2, and -kappaB3, according to their electropho retic mobilities (fast, intermediate, and slow, respectively). Immunol ogical and UV cross-linking studies showed that NF-kappaB1 consists of only p50 subunit, whereas both NF-kappaB2 and -kappaB3 contain NF-kap paB p65 subunit and c-Rel. In addition, NF-kappaB2 was also found to c ontain p50 subunit of NF-kappaB. The binding of three types of NF-kapp aB proteins to HIV NF-kappaB motif was effectively inhibited by other NF-kappaB motifs, whose 3' half-site nucleotide sequences are T/A-T-T/ C-CC (HIV, interleukin-6, interferon (INF)-beta, H2-K(b), I-Ealpha(d), and TNF-alpha2 (nucleotide position -510)) and much less effectively by NF-kappaB motifs with 3' half-site sequences of TGCCC (TNF-alpha3, nucleotide position -610), ATCTC (G-CSF), TATTC (FcgammaR), or TCCTT ( TNF-alpha1, nucleotide position -850). Our data also suggested that NF -kappaB1 and -kappaB2 which contain p50 subunit of NF-kappaB bind with the higher preference for NF-kappaB motif of H2-K(b) gene promoter th an that of INF-beta, whereas NF-kappaB3 which does not contain p50 sub unit appears to preferentially bind to NF-kappaB sites of IFN-beta.