High-throughput AFM analysis reveals unwrapping pathways of H3 and CENP-A nucleosomes
| Autor: | Konrad, Sebastian F., Vanderlinden, Willem, Frederickx, Wout, Brouns, Tine, Menze, Björn H., De Feyter, Steven, Lipfert, Jan, Sub Molecular Biophysics, Soft Condensed Matter and Biophysics |
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| Přispěvatelé: | University of Zurich, Vanderlinden, Willem, Lipfert, Jan, Sub Molecular Biophysics, Soft Condensed Matter and Biophysics |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Physics
0303 health sciences Atomic force microscopy 030302 biochemistry & molecular biology Centromere 610 Medicine & health Computational biology 2500 General Materials Science Chromatin Nucleoprotein Epigenesis Genetic Nucleosomes Histones 03 medical and health sciences Nucleosome General Materials Science Throughput (business) 11493 Department of Quantitative Biomedicine Centromere Protein A 030304 developmental biology |
| Zdroj: | Nanoscale, 13(10), 5435. Royal Society of Chemistry |
| ISSN: | 2040-3364 |
| DOI: | 10.1101/2020.04.09.034090 |
| Popis: | Nucleosomes, the fundamental units of chromatin, regulate readout and expression of eukaryotic genomes. Single-molecule experiments have revealed force-induced nucleosome accessibility, but a high-resolution unwrapping landscape in the absence of external forces is currently lacking. Here, we introduce a high-throughput pipeline for the analysis of nucleosome conformations based on atomic force microscopy and automated, multi-parameter image analysis. Our data set of ∼10 000 nucleosomes reveals multiple unwrapping states corresponding to steps of 5 bp DNA. For canonical H3 nucleosomes, we observe that dissociation from one side impedes unwrapping from the other side, but in contrast to force-induced unwrapping, we find only a weak sequence-dependent asymmetry. Notably, centromeric CENP-A nucleosomes do not unwrap anti-cooperatively, in stark contrast to H3 nucleosomes. Finally, our results reconcile previous conflicting findings about the differences in height between H3 and CENP-A nucleosomes. We expect our approach to enable critical insights into epigenetic regulation of nucleosome structure and stability and to facilitate future high-throughput AFM studies that involve heterogeneous nucleoprotein complexes. ispartof: NANOSCALE vol:13 issue:10 pages:5435-5447 ispartof: location:England status: published |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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