Reconstituted TAD-size chromatin fibers feature heterogeneous nucleosome clusters
Large topologically associated domains (TADs) contain irregularly spaced nucleosome clutches, and interactions between such clutches are thought to aid the compaction of these domains. Here, we reconstituted TAD-sized chromatin fibers containing hundreds of nucleosomes on native source human and lam...
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sg-ntu-dr.10356-1654512023-03-27T15:31:57Z Reconstituted TAD-size chromatin fibers feature heterogeneous nucleosome clusters Korolev, Nikolay Zinchenko, Anatoly Soman, Aghil Chen, Qinming Wong, Sook Yi Berezhnoy, Nikolay V. Basak, Rajib van der Maarel, Johan R. C. van Noort, John Nordenskiöld, Lars School of Biological Sciences Science::Biological sciences Histone Chromatin Large topologically associated domains (TADs) contain irregularly spaced nucleosome clutches, and interactions between such clutches are thought to aid the compaction of these domains. Here, we reconstituted TAD-sized chromatin fibers containing hundreds of nucleosomes on native source human and lambda-phage DNA and compared their mechanical properties at the single-molecule level with shorter '601' arrays with various nucleosome repeat lengths. Fluorescent imaging showed increased compaction upon saturation of the DNA with histones and increasing magnesium concentration. Nucleosome clusters and their structural fluctuations were visualized in confined nanochannels. Force spectroscopy revealed not only similar mechanical properties of the TAD-sized fibers as shorter fibers but also large rupture events, consistent with breaking the interactions between distant clutches of nucleosomes. Though the arrays of native human DNA, lambda-phage and '601' DNA featured minor differences in reconstitution yield and nucleosome stability, the fibers' global structural and mechanical properties were similar, including the interactions between nucleosome clutches. These single-molecule experiments quantify the mechanical forces that stabilize large TAD-sized chromatin domains consisting of disordered, dynamically interacting nucleosome clutches and their effect on the condensation of large chromatin domains. Ministry of Education (MOE) Published version This work was supported by the Singapore Ministry of Education (MOE) Academic Research Fund (AcRF) Tier 1 [2018-T1-001-114 to L.N.] and Tier 3 [2012-T3-1-001 to L.N.] grants. This research was also supported by the Singapore Ministry of Education Academic Research Fund Tier 1 Grant R-144-000-451-114 and Tier 2 grant MOE-T2EP50121-0003 to JvdM. 2023-03-27T07:56:10Z 2023-03-27T07:56:10Z 2022 Journal Article Korolev, N., Zinchenko, A., Soman, A., Chen, Q., Wong, S. Y., Berezhnoy, N. V., Basak, R., van der Maarel, J. R. C., van Noort, J. & Nordenskiöld, L. (2022). Reconstituted TAD-size chromatin fibers feature heterogeneous nucleosome clusters. Scientific Reports, 12(1), 15558-. https://dx.doi.org/10.1038/s41598-022-19471-3 2045-2322 https://hdl.handle.net/10356/165451 10.1038/s41598-022-19471-3 36114220 2-s2.0-85138151899 1 12 15558 en 2018-T1-001-114 2012-T3-1-001 Scientific Reports © The Author(s) 2022. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. application/pdf |
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Science::Biological sciences Histone Chromatin Korolev, Nikolay Zinchenko, Anatoly Soman, Aghil Chen, Qinming Wong, Sook Yi Berezhnoy, Nikolay V. Basak, Rajib van der Maarel, Johan R. C. van Noort, John Nordenskiöld, Lars Reconstituted TAD-size chromatin fibers feature heterogeneous nucleosome clusters |
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Large topologically associated domains (TADs) contain irregularly spaced nucleosome clutches, and interactions between such clutches are thought to aid the compaction of these domains. Here, we reconstituted TAD-sized chromatin fibers containing hundreds of nucleosomes on native source human and lambda-phage DNA and compared their mechanical properties at the single-molecule level with shorter '601' arrays with various nucleosome repeat lengths. Fluorescent imaging showed increased compaction upon saturation of the DNA with histones and increasing magnesium concentration. Nucleosome clusters and their structural fluctuations were visualized in confined nanochannels. Force spectroscopy revealed not only similar mechanical properties of the TAD-sized fibers as shorter fibers but also large rupture events, consistent with breaking the interactions between distant clutches of nucleosomes. Though the arrays of native human DNA, lambda-phage and '601' DNA featured minor differences in reconstitution yield and nucleosome stability, the fibers' global structural and mechanical properties were similar, including the interactions between nucleosome clutches. These single-molecule experiments quantify the mechanical forces that stabilize large TAD-sized chromatin domains consisting of disordered, dynamically interacting nucleosome clutches and their effect on the condensation of large chromatin domains. |
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School of Biological Sciences |
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School of Biological Sciences Korolev, Nikolay Zinchenko, Anatoly Soman, Aghil Chen, Qinming Wong, Sook Yi Berezhnoy, Nikolay V. Basak, Rajib van der Maarel, Johan R. C. van Noort, John Nordenskiöld, Lars |
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Article |
author |
Korolev, Nikolay Zinchenko, Anatoly Soman, Aghil Chen, Qinming Wong, Sook Yi Berezhnoy, Nikolay V. Basak, Rajib van der Maarel, Johan R. C. van Noort, John Nordenskiöld, Lars |
author_sort |
Korolev, Nikolay |
title |
Reconstituted TAD-size chromatin fibers feature heterogeneous nucleosome clusters |
title_short |
Reconstituted TAD-size chromatin fibers feature heterogeneous nucleosome clusters |
title_full |
Reconstituted TAD-size chromatin fibers feature heterogeneous nucleosome clusters |
title_fullStr |
Reconstituted TAD-size chromatin fibers feature heterogeneous nucleosome clusters |
title_full_unstemmed |
Reconstituted TAD-size chromatin fibers feature heterogeneous nucleosome clusters |
title_sort |
reconstituted tad-size chromatin fibers feature heterogeneous nucleosome clusters |
publishDate |
2023 |
url |
https://hdl.handle.net/10356/165451 |
_version_ |
1761781698492628992 |