Disordered regions and folded modules in CAF-1 promote histone deposition in Schizosaccharomyces pombe

Genome and epigenome integrity in eukaryotes depends on the proper coupling of histone deposition with DNA synthesis. This process relies on the evolutionary conserved histone chaperone CAF-1 for which the links between structure and functions are still a puzzle. While studies of the Saccharomyces c...

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Main Authors: Fouad Ouasti, Maxime Audin, Karine Fréon, Jean-Pierre Quivy, Mehdi Tachekort, Elizabeth Cesard, Aurélien Thureau, Virginie Ropars, Paloma Fernández Varela, Gwenaelle Moal, Ibrahim Soumana Adamou, Aleksandra Uryga, Pierre Legrand, Jessica Andreani, Raphaël Guerois, Geneviève Almouzni, Sarah Lambert, Francoise Ochsenbein
Format: Article
Language:English
Published: eLife Sciences Publications Ltd 2024-02-01
Series:eLife
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Online Access:https://elifesciences.org/articles/91461
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author Fouad Ouasti
Maxime Audin
Karine Fréon
Jean-Pierre Quivy
Mehdi Tachekort
Elizabeth Cesard
Aurélien Thureau
Virginie Ropars
Paloma Fernández Varela
Gwenaelle Moal
Ibrahim Soumana Adamou
Aleksandra Uryga
Pierre Legrand
Jessica Andreani
Raphaël Guerois
Geneviève Almouzni
Sarah Lambert
Francoise Ochsenbein
author_facet Fouad Ouasti
Maxime Audin
Karine Fréon
Jean-Pierre Quivy
Mehdi Tachekort
Elizabeth Cesard
Aurélien Thureau
Virginie Ropars
Paloma Fernández Varela
Gwenaelle Moal
Ibrahim Soumana Adamou
Aleksandra Uryga
Pierre Legrand
Jessica Andreani
Raphaël Guerois
Geneviève Almouzni
Sarah Lambert
Francoise Ochsenbein
author_sort Fouad Ouasti
collection DOAJ
description Genome and epigenome integrity in eukaryotes depends on the proper coupling of histone deposition with DNA synthesis. This process relies on the evolutionary conserved histone chaperone CAF-1 for which the links between structure and functions are still a puzzle. While studies of the Saccharomyces cerevisiae CAF-1 complex enabled to propose a model for the histone deposition mechanism, we still lack a framework to demonstrate its generality and in particular, how its interaction with the polymerase accessory factor PCNA is operating. Here, we reconstituted a complete SpCAF-1 from fission yeast. We characterized its dynamic structure using NMR, SAXS and molecular modeling together with in vitro and in vivo functional studies on rationally designed interaction mutants. Importantly, we identify the unfolded nature of the acidic domain which folds up when binding to histones. We also show how the long KER helix mediates DNA binding and stimulates SpCAF-1 association with PCNA. Our study highlights how the organization of CAF-1 comprising both disordered regions and folded modules enables the dynamics of multiple interactions to promote synthesis-coupled histone deposition essential for its DNA replication, heterochromatin maintenance, and genome stability functions.
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spelling doaj-art-e4eaadc73ce04cfaa4c47752fee0fda92025-08-20T02:26:20ZengeLife Sciences Publications LtdeLife2050-084X2024-02-011210.7554/eLife.91461Disordered regions and folded modules in CAF-1 promote histone deposition in Schizosaccharomyces pombeFouad Ouasti0Maxime Audin1Karine Fréon2https://orcid.org/0000-0001-7853-078XJean-Pierre Quivy3https://orcid.org/0000-0001-6557-7204Mehdi Tachekort4Elizabeth Cesard5Aurélien Thureau6Virginie Ropars7Paloma Fernández Varela8Gwenaelle Moal9Ibrahim Soumana Adamou10Aleksandra Uryga11Pierre Legrand12Jessica Andreani13https://orcid.org/0000-0003-4435-9093Raphaël Guerois14https://orcid.org/0000-0001-5294-2858Geneviève Almouzni15Sarah Lambert16https://orcid.org/0000-0002-1403-3204Francoise Ochsenbein17https://orcid.org/0000-0002-9027-4384Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceInstitut Curie, PSL Research University, CNRS UMR 3348, INSERM U1278, Université Paris-Saclay, Equipe labellisée Ligue contre le Cancer, Orsay, FranceInstitut Curie, PSL Research University, CNRS, Sorbonne Université,CNRS UMR3664, Nuclear Dynamics Unit, Équipe Labellisée Ligue contre le Cancer, Paris, FranceUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceSynchrotron SOLEIL, HelioBio group, l'Orme des Merisiers, Saint-Aubin, FranceUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceInstitut Curie, PSL Research University, CNRS UMR 3348, INSERM U1278, Université Paris-Saclay, Equipe labellisée Ligue contre le Cancer, Orsay, FranceInstitut Curie, PSL Research University, CNRS UMR 3348, INSERM U1278, Université Paris-Saclay, Equipe labellisée Ligue contre le Cancer, Orsay, FranceSynchrotron SOLEIL, HelioBio group, l'Orme des Merisiers, Saint-Aubin, FranceUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceInstitut Curie, PSL Research University, CNRS, Sorbonne Université,CNRS UMR3664, Nuclear Dynamics Unit, Équipe Labellisée Ligue contre le Cancer, Paris, FranceInstitut Curie, PSL Research University, CNRS UMR 3348, INSERM U1278, Université Paris-Saclay, Equipe labellisée Ligue contre le Cancer, Orsay, FranceUniversité Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Institute Joliot, Gif-sur-Yvette, FranceGenome and epigenome integrity in eukaryotes depends on the proper coupling of histone deposition with DNA synthesis. This process relies on the evolutionary conserved histone chaperone CAF-1 for which the links between structure and functions are still a puzzle. While studies of the Saccharomyces cerevisiae CAF-1 complex enabled to propose a model for the histone deposition mechanism, we still lack a framework to demonstrate its generality and in particular, how its interaction with the polymerase accessory factor PCNA is operating. Here, we reconstituted a complete SpCAF-1 from fission yeast. We characterized its dynamic structure using NMR, SAXS and molecular modeling together with in vitro and in vivo functional studies on rationally designed interaction mutants. Importantly, we identify the unfolded nature of the acidic domain which folds up when binding to histones. We also show how the long KER helix mediates DNA binding and stimulates SpCAF-1 association with PCNA. Our study highlights how the organization of CAF-1 comprising both disordered regions and folded modules enables the dynamics of multiple interactions to promote synthesis-coupled histone deposition essential for its DNA replication, heterochromatin maintenance, and genome stability functions.https://elifesciences.org/articles/91461histone chaperoneNucleosome assemblyepigeneticgenome integritynmrsaxs
spellingShingle Fouad Ouasti
Maxime Audin
Karine Fréon
Jean-Pierre Quivy
Mehdi Tachekort
Elizabeth Cesard
Aurélien Thureau
Virginie Ropars
Paloma Fernández Varela
Gwenaelle Moal
Ibrahim Soumana Adamou
Aleksandra Uryga
Pierre Legrand
Jessica Andreani
Raphaël Guerois
Geneviève Almouzni
Sarah Lambert
Francoise Ochsenbein
Disordered regions and folded modules in CAF-1 promote histone deposition in Schizosaccharomyces pombe
eLife
histone chaperone
Nucleosome assembly
epigenetic
genome integrity
nmr
saxs
title Disordered regions and folded modules in CAF-1 promote histone deposition in Schizosaccharomyces pombe
title_full Disordered regions and folded modules in CAF-1 promote histone deposition in Schizosaccharomyces pombe
title_fullStr Disordered regions and folded modules in CAF-1 promote histone deposition in Schizosaccharomyces pombe
title_full_unstemmed Disordered regions and folded modules in CAF-1 promote histone deposition in Schizosaccharomyces pombe
title_short Disordered regions and folded modules in CAF-1 promote histone deposition in Schizosaccharomyces pombe
title_sort disordered regions and folded modules in caf 1 promote histone deposition in schizosaccharomyces pombe
topic histone chaperone
Nucleosome assembly
epigenetic
genome integrity
nmr
saxs
url https://elifesciences.org/articles/91461
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