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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...

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Autores principales: Shintomi, Keishi, Hirano, Tatsuya
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
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.
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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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