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bioRxiv · 10.1101/2024.02.13.580208

The Interaction of NF-κB Transcription Factor with Centromeric Chromatin

Filliaux, S. B.·Bertelsen, C.·Baughman, H. E. R.·Komives, E. A. A.·Lyubchenko, Y. L.
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Abstract

Centromeric chromatin is a subset of chromatin structure and governs chromosome segregation. The centromere is composed of both CENP-A nucleosomes (CENP-Anuc) and H3 nucleosomes (H3nuc) and is enriched with alpha-satellite (-sat) DNA repeats. These CENP-Anuc have a different structure than H3nuc, decreasing the base pairs (bp) of wrapped DNA from 147 bp for H3nuc to 121 bp for CENP-Anuc. All these factors can contribute to centromere function. We investigated the interaction of H3nuc and CENP-Anuc with NF-{kappa}B, 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-{kappa}B. We found that NF-{kappa}B unravels H3nuc, removing more than 20 bp of DNA, and that NF-{kappa}B binds to the nucleosomal core. Similar results were obtained for the truncated variant of NF-{kappa}B comprised only of the Rel Homology domain and missing the transcription activation domain (TAD), suggesting the RelA TAD is not critical in unraveling H3nuc. By contrast, NF-{kappa}B did not bind to or unravel CENP- Anuc. These findings with different affinities for two types of nucleosomes to NF-{kappa}B may have implications for understanding the mechanisms of gene expression in bulk and centromere chromatin.

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Filliaux, S. B., Bertelsen, C., Baughman, H. E. R., Komives, E. A. A., Lyubchenko, Y. L.. 2024-02-16. The Interaction of NF-κB Transcription Factor with Centromeric Chromatin. https://doi.org/10.1101/2024.02.13.580208

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