Mechanism of kappa B DNA binding by Rel/NF-kappa B dimers

The DNA binding of three different NF-kappaB dimers, the p50 and p65 homodimers and the p50/p65 heterodimer, has been examined using a combination of gel mobility shift and fluorescence anisotropy assays. The NF-kappaB p50/p65 heterodimer is shown here to bind the kappaB DNA target site of the immunoglobulin kappa enhancer (Ig-kappaB) with an affinity of approximately 10 nm. The p50 and p65 homodimers bind to the same site with roughly 5- and 15-fold lower affinity, respectively. The nature of the binding isotherms indicates a cooperative mode of binding for all three dimers to the DNA targets. We have further characterized the role of pH, salt, and temperature on the formation of the p50/p65 heterodimer-Ig-kappaB complex. The heterodimer binds to the Ig-kappaB DNA target in a pH-dependent manner, with the highest affinity between pH 7.0 and 7.5. A strong salt-dependent interaction between Ig-kappaB and the p50/p65 heterodimer is observed, with optimum binding occurring at monovalent salt concentrations below 75 mm, with binding becoming virtually nonspecific at a salt concentration of 200 mm. Binding of the heterodimer to DNA was unchanged across a temperature range between 4 degrees C and 42 degrees C. The sensitivity to ionic environment and insensitivity to temperature indicate that NF-kappaB p50/p65 heterodimers form complexes with specific DNA in an entropically driven manner.     

Expression of LMP1 in epithelial cells leads to the activation of a select subset of NF-kappa B/Rel family proteins.

This study demonstrates that the Epstein-Barr virus protein LMP1 activates a specific subset of NF-kappa B/Rel proteins in the C33 epithelial cell line. Western immunoblot analysis used to analyze the intracellular distribution and abundance of the proteins present in these complexes demonstrated that levels of the p50 and p52 proteins were significantly elevated in the nuclei of LMP1-expressing cells. The data also suggest that LMP1 facilitates the translocation of p50 to the nucleus and may affect the processing of the p100 and p105 precursor proteins or the stability of p52 and p50.     

Sequence homologies amongE. coli ribosomal proteins: Evidence for evolutionarily related groupings and internal duplications

Summary The complete or partial sequences of 47E. coli ribosomal proteins described in the literature have been examined by computerized search and matching programs. In contrast to results previously reported by other investigators, sequence homologies were uncovered among some of these ribosomal proteins that are well beyond statistical expectations. Moreover, alignments of the most strongly homologous sequences suggested the existence of a network of family groupings. Several of these proteins also exhibit internal homologies, indicating that they have been elongated by a series of tandem duplications.     

Sarcophagid larval proteins: Partial sequence homologies among three cuticle proteins and related structures of drosophilids

Summary Three structural proteins from the larval cuticle ofSarcophaga bullata have been sequenced at the amino terminus for 30–40 residues. We observed a high degree of homology with related proteins ofDrosophila melanogaster, based on the previous findings of M. Snyder, J. Hirsh, and N. Davidson [(1981) Cell 25:165–177].S. bullata protein SC1 had 65% homology withDrosophila isolate CP1, and SC6 showed 49% homology with CPX and 54% with CP2a. The three sarcophagid polypeptides also resembled each other with respect to mapped products of tryptic cleavage. The sites of posttranslational arylation required for puparium formation, namely histidyl and lysyl residues, were asymmetrically distributed in the sarcophagid samples. In SC1 the bulk of the loci of putative crosslinks lay beyond the 43-residue fragment. In SC6 half the histidines fell within the first 25% of the primary chain.     

The inhibitory ankyrin and activator Rel proteins.

The gene families encoding the proteins NF-kappa B, c-Rel and Dorsal, in conjunction with their respective inhibitors l kappa B, pp40, and Cactus, achieve specificity in gene regulation by means of common principles. The related activities of NF-kappa B and Dorsal are mediated by heterodimeric or homodimeric complexes of proteins containing the conserved dimerization and DNA-binding domain termed Rel. The l kappa Bs and Cactus, which share a core series of structural repeats termed ankyrin, inhibit cognate activators through differential interactions with the Rel-homology domain. Together, the inhibitory ankyrin proteins and their cognate Rel dimers probably define specific signalling pathways able to activate specific gene expression. Both gene families include proto-oncogenes, thus broadly implicating Rel/l kappa B in the control of both normal gene expression and the aberrant gene expression that makes cells cancerous.