Heterodimerization and transcriptional activation in vitro by NF‐κB proteins

Abstract

The NF-kappa B family of transcription proteins represents multiple DNA binding, rel related polypeptides that contribute to regulation of genes involved in immune responsiveness and inflammation, as well as activation of the HIV long terminal repeat. In this study multiple NF-kappa B related polypeptides ranging from 85 to 45 kDa were examined for their capacity to interact with the PRDII regulatory element of interferon beta and were shown to possess distinct intrinsic DNA binding affinities for this NF-kappa B site and form multiple DNA binding homo- and heterodimer complexes in co-renaturation experiments. Furthermore, using DNA templates containing two copies of the PRDII domain linked to the rabbit beta globin gene, the purified polypeptides specifically stimulated NF-kappa B dependent transcription in an in vitro reconstitution assay as heterodimers but not as p50 homodimers. These experiments emphasize the role of NF-kappa B dimerization as a distinct level of transcriptional control that may permit functional diversification of a limited number of regulatory proteins.

Characterization of a functional NF-kappa B site in the human interleukin 1 beta promoter: evidence for a positive autoregulatory loop.

“…Cell extracts were prepared essentially as described previously (34,35). A 5-to 15-p,g sample of WCE was preincubated with 5 p,g of poly(dI-dC) in nuclear dialysis buffer (14) for 10 min at 4°C. AACGTGGGAAA-3' (NF-KB oligonucleotide).…”

Viral induction of the human beta interferon promoter: modulation of transcription by NF-kappa B/rel proteins and interferon regulatory factors

“…The second positive regulatory domain of IFN-1, PRDII (-64 to -55), is a decamer sequence (5'-GGGAAATTCC-3') that serves as a recognition site for the NF-KB/rel family of transcription factors (10,17,31,41,72). Base mutations or deletions which alter the PRDII domain cause a loss of virus inducibility, suggesting a critical role for this element in virus induction (23,24,71).…”

Cell-penetrating interactomic inhibition of nuclear factor-kappa B in a mouse model of postoperative cognitive dysfunction

“…Inhibitory IκB binds to NF-κB as an inactive cytoplasmic complex and controls the DNA-binding activity of NF-κB by translocating it to the nucleus 32 , 35 . However, translocated nuclear NF-κB participates in the activation of immune cells and controls genes that are associated with inflammatory cytokines, chemokines, cell adhesion molecules, and major histocompatibility complex proteins 14 , 15 , 36 .…”