Centromeres are maintained by fastening CENP-A to DNA and directing an arginine anchor-dependent nucleosome transition

• Published in: Nature communications Vol. 8; no. 1; p. 15775
• Main Authors: Guo, Lucie Y., Allu, Praveen Kumar, Zandarashvili, Levani, McKinley, Kara L., Sekulic, Nikolina, Dawicki-McKenna, Jennine M., Fachinetti, Daniele, Logsdon, Glennis A., Jamiolkowski, Ryan M., Cleveland, Don W., Cheeseman, Iain M., Black, Ben E.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 09.06.2017; Nature Publishing Group; Nature Portfolio
• Subjects: 101/58; 45/41; 45/70; 631/57/2272/2276; 631/80/103/90; 82/83; 96/106; 96/109; 96/63; Animals; Arginine - metabolism; Biochemistry; Biophysics; Cell cycle; Centromere - chemistry; Centromere - genetics; Centromere - metabolism; Centromere Protein A - chemistry; Centromere Protein A - genetics; Centromere Protein A - metabolism; Chromosomal Proteins, Non-Histone - genetics; Chromosomal Proteins, Non-Histone - metabolism; Chromosomes; DNA - genetics; DNA - metabolism; Epigenetics; Female; Humanities and Social Sciences; Humans; Male; Medical research; Medicine; Mice; multidisciplinary; Nucleosomes - chemistry; Nucleosomes - genetics; Nucleosomes - metabolism; Protein Binding; Protein Domains; Proteins; Science; Science (multidisciplinary); California; United States > US; Massachusetts; Pennsylvania
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms15775

Maintaining centromere identity relies upon the persistence of the epigenetic mark provided by the histone H3 variant, centromere protein A (CENP-A), but the molecular mechanisms that underlie its remarkable stability remain unclear. Here, we define the contributions of each of the three candidate CENP-A nucleosome-binding domains (two on CENP-C and one on CENP-N) to CENP-A stability using gene replacement and rapid protein degradation. Surprisingly, the most conserved domain, the CENP-C motif, is dispensable. Instead, the stability is conferred by the unfolded central domain of CENP-C and the folded N-terminal domain of CENP-N that becomes rigidified 1,000-fold upon crossbridging CENP-A and its adjacent nucleosomal DNA. Disrupting the ‘arginine anchor’ on CENP-C for the nucleosomal acidic patch disrupts the CENP-A nucleosome structural transition and removes CENP-A nucleosomes from centromeres. CENP-A nucleosome retention at centromeres requires a core centromeric nucleosome complex where CENP-C clamps down a stable nucleosome conformation and CENP-N fastens CENP-A to the DNA. Centromere maintenance depends on the persistence of the histone variant CENP-A at the centromeres. Here, the authors characterize the core centromeric nucleosome complex wherein CENP-C confers a stable CENP-A nucleosome conformation and CENP-N fastens CENP-A to the DNA.