The Interaction of NF-κB Transcription Factor with Centromeric Chromatin.

Centromeric chromatin is a subset of chromatin structure and governs chromosome segregation. The centromere is composed of both CENP-A nucleosomes (CENP-A ) and H3 nucleosomes (H3 ) and is enriched with alpha-satellite (α-sat) DNA repeats. These CENP-A have a different structure than H3 , decreasing the base pairs (bp) of wrapped DNA from 147 bp for H3 to 121 bp for CENP-A . All these factors can contribute to centromere function. We investigated the interaction of H3 and CENP-A with NF-κB, a crucial transcription factor in regulating immune response and inflammation. We utilized Atomic Force Microscopy (AFM) to characterize complexes of both types of nucleosomes with NF-κB. We found that NF-κB unravels H3 , removing more than 20 bp of DNA, and that NF-κB binds to the nucleosomal core. Similar results were obtained for the truncated variant of NF-κB comprised only of the Rel Homology domain and missing the transcription activation domain (TAD), suggesting the RelA TAD is not critical in unraveling H3 . By contrast, NF-κB did not bind to or unravel CENP- A . These findings with different affinities for two types of nucleosomes to NF-κB may have implications for understanding the mechanisms of gene expression in bulk and centromere chromatin.