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Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA

Cohesin is a fundamental protein complex that holds sister chromatids together. Separase protease cleaves a cohesin subunit Rad21/SCC1, causing the release of cohesin from DNA to allow chromosome segregation. To understand the functional organization of cohesin, we employed next-generation whole-gen...

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Autores principales: Xu, Xingya, Kanai, Ryuta, Nakazawa, Norihiko, Wang, Li, Toyoshima, Chikashi, Yanagida, Mitsuhiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6003501/
https://www.ncbi.nlm.nih.gov/pubmed/29735656
http://dx.doi.org/10.1073/pnas.1803564115
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author Xu, Xingya
Kanai, Ryuta
Nakazawa, Norihiko
Wang, Li
Toyoshima, Chikashi
Yanagida, Mitsuhiro
author_facet Xu, Xingya
Kanai, Ryuta
Nakazawa, Norihiko
Wang, Li
Toyoshima, Chikashi
Yanagida, Mitsuhiro
author_sort Xu, Xingya
collection PubMed
description Cohesin is a fundamental protein complex that holds sister chromatids together. Separase protease cleaves a cohesin subunit Rad21/SCC1, causing the release of cohesin from DNA to allow chromosome segregation. To understand the functional organization of cohesin, we employed next-generation whole-genome sequencing and identified numerous extragenic suppressors that overcome either inactive separase/Cut1 or defective cohesin in the fission yeast Schizosaccharomyces pombe. Unexpectedly, Cut1 is dispensable if suppressor mutations cause disorders of interfaces among essential cohesin subunits Psm1/SMC1, Psm3/SMC3, Rad21/SCC1, and Mis4/SCC2, the crystal structures of which suggest physical and functional impairment at the interfaces of Psm1/3 hinge, Psm1 head–Rad21, or Psm3 coiled coil–Rad21. Molecular-dynamics analysis indicates that the intermolecular β-sheets in the cohesin hinge of cut1 suppressor mutants remain intact, but a large mobility change occurs at the coiled coil bound to the hinge. In contrast, suppressors of rad21-K1 occur in either the head ATPase domains or the Psm3 coiled coil that interacts with Rad21. Suppressors of mis4-G1326E reside in the head of Psm3/1 or the intragenic domain of Mis4. These may restore the binding of cohesin to DNA. Evidence is provided that the head and hinge of SMC subunits are proximal, and that they coordinate to form arched coils that can hold or release DNA by altering the angles made by the arched coiled coils. By combining molecular modeling with suppressor sequence analysis, we propose a cohesin structure designated the “hold-and-release” model, which may be considered as an alternative to the prevailing “ring” model.
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spelling pubmed-60035012018-06-18 Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA Xu, Xingya Kanai, Ryuta Nakazawa, Norihiko Wang, Li Toyoshima, Chikashi Yanagida, Mitsuhiro Proc Natl Acad Sci U S A PNAS Plus Cohesin is a fundamental protein complex that holds sister chromatids together. Separase protease cleaves a cohesin subunit Rad21/SCC1, causing the release of cohesin from DNA to allow chromosome segregation. To understand the functional organization of cohesin, we employed next-generation whole-genome sequencing and identified numerous extragenic suppressors that overcome either inactive separase/Cut1 or defective cohesin in the fission yeast Schizosaccharomyces pombe. Unexpectedly, Cut1 is dispensable if suppressor mutations cause disorders of interfaces among essential cohesin subunits Psm1/SMC1, Psm3/SMC3, Rad21/SCC1, and Mis4/SCC2, the crystal structures of which suggest physical and functional impairment at the interfaces of Psm1/3 hinge, Psm1 head–Rad21, or Psm3 coiled coil–Rad21. Molecular-dynamics analysis indicates that the intermolecular β-sheets in the cohesin hinge of cut1 suppressor mutants remain intact, but a large mobility change occurs at the coiled coil bound to the hinge. In contrast, suppressors of rad21-K1 occur in either the head ATPase domains or the Psm3 coiled coil that interacts with Rad21. Suppressors of mis4-G1326E reside in the head of Psm3/1 or the intragenic domain of Mis4. These may restore the binding of cohesin to DNA. Evidence is provided that the head and hinge of SMC subunits are proximal, and that they coordinate to form arched coils that can hold or release DNA by altering the angles made by the arched coiled coils. By combining molecular modeling with suppressor sequence analysis, we propose a cohesin structure designated the “hold-and-release” model, which may be considered as an alternative to the prevailing “ring” model. National Academy of Sciences 2018-05-22 2018-05-07 /pmc/articles/PMC6003501/ /pubmed/29735656 http://dx.doi.org/10.1073/pnas.1803564115 Text en Copyright © 2018 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/ This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle PNAS Plus
Xu, Xingya
Kanai, Ryuta
Nakazawa, Norihiko
Wang, Li
Toyoshima, Chikashi
Yanagida, Mitsuhiro
Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA
title Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA
title_full Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA
title_fullStr Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA
title_full_unstemmed Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA
title_short Suppressor mutation analysis combined with 3D modeling explains cohesin’s capacity to hold and release DNA
title_sort suppressor mutation analysis combined with 3d modeling explains cohesin’s capacity to hold and release dna
topic PNAS Plus
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6003501/
https://www.ncbi.nlm.nih.gov/pubmed/29735656
http://dx.doi.org/10.1073/pnas.1803564115
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