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Guiding functions of the C-terminal domain of topoisomerase IIα advance mitotic chromosome assembly
Topoisomerase II (topo II) is one of the six proteins essential for mitotic chromatid reconstitution in vitro. It is not fully understood, however, mechanistically how this enzyme regulates this process. In an attempt to further refine the reconstitution assay, we have found that chromosomal binding...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8131626/ https://www.ncbi.nlm.nih.gov/pubmed/34006877 http://dx.doi.org/10.1038/s41467-021-23205-w |
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author | Shintomi, Keishi Hirano, Tatsuya |
author_facet | Shintomi, Keishi Hirano, Tatsuya |
author_sort | Shintomi, Keishi |
collection | PubMed |
description | Topoisomerase II (topo II) is one of the six proteins essential for mitotic chromatid reconstitution in vitro. It is not fully understood, however, mechanistically how this enzyme regulates this process. In an attempt to further refine the reconstitution assay, we have found that chromosomal binding of Xenopus laevis topo IIα is sensitive to buffer conditions and depends on its C-terminal domain (CTD). Enzymological assays using circular DNA substrates supports the idea that topo IIα first resolves inter-chromatid entanglements to drive individualization and then generates intra-chromatid entanglements to promote thickening. Importantly, only the latter process requires the CTD. By using frog egg extracts, we also show that the CTD contributes to proper formation of nucleosome-depleted chromatids by competing with a linker histone for non-nucleosomal DNA. Our results demonstrate that topo IIα utilizes its CTD to deliver the enzymatic core to crowded environments created during mitotic chromatid assembly, thereby fine-tuning this process. |
format | Online Article Text |
id | pubmed-8131626 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-81316262021-05-24 Guiding functions of the C-terminal domain of topoisomerase IIα advance mitotic chromosome assembly Shintomi, Keishi Hirano, Tatsuya Nat Commun Article Topoisomerase II (topo II) is one of the six proteins essential for mitotic chromatid reconstitution in vitro. It is not fully understood, however, mechanistically how this enzyme regulates this process. In an attempt to further refine the reconstitution assay, we have found that chromosomal binding of Xenopus laevis topo IIα is sensitive to buffer conditions and depends on its C-terminal domain (CTD). Enzymological assays using circular DNA substrates supports the idea that topo IIα first resolves inter-chromatid entanglements to drive individualization and then generates intra-chromatid entanglements to promote thickening. Importantly, only the latter process requires the CTD. By using frog egg extracts, we also show that the CTD contributes to proper formation of nucleosome-depleted chromatids by competing with a linker histone for non-nucleosomal DNA. Our results demonstrate that topo IIα utilizes its CTD to deliver the enzymatic core to crowded environments created during mitotic chromatid assembly, thereby fine-tuning this process. Nature Publishing Group UK 2021-05-18 /pmc/articles/PMC8131626/ /pubmed/34006877 http://dx.doi.org/10.1038/s41467-021-23205-w Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Shintomi, Keishi Hirano, Tatsuya Guiding functions of the C-terminal domain of topoisomerase IIα advance mitotic chromosome assembly |
title | Guiding functions of the C-terminal domain of topoisomerase IIα advance mitotic chromosome assembly |
title_full | Guiding functions of the C-terminal domain of topoisomerase IIα advance mitotic chromosome assembly |
title_fullStr | Guiding functions of the C-terminal domain of topoisomerase IIα advance mitotic chromosome assembly |
title_full_unstemmed | Guiding functions of the C-terminal domain of topoisomerase IIα advance mitotic chromosome assembly |
title_short | Guiding functions of the C-terminal domain of topoisomerase IIα advance mitotic chromosome assembly |
title_sort | guiding functions of the c-terminal domain of topoisomerase iiα advance mitotic chromosome assembly |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8131626/ https://www.ncbi.nlm.nih.gov/pubmed/34006877 http://dx.doi.org/10.1038/s41467-021-23205-w |
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