An Inducible System for Silencing Establishment Reveals a Stepwise Mechanism in Which Anchoring at the Nuclear Periphery Precedes Heterochromatin Formation

In eukaryotic cells, silent chromatin is mainly found at the nuclear periphery forming subnuclear compartments that favor silencing establishment. Here, we set up an inducible system to monitor silencing establishment at an ectopic locus in relation with its subnuclear localization in budding yeast....

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Autores principales:	Isabelle Loïodice, Mickael Garnier, Ivaylo Nikolov, Angela Taddei
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	chromatin heterochromatin nuclear organization gene regulation gene silencing FROS Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/fc8e847ee24e4172ae27a15b8d00532b
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spelling	oai:doaj.org-article:fc8e847ee24e4172ae27a15b8d00532b2021-11-25T17:07:23ZAn Inducible System for Silencing Establishment Reveals a Stepwise Mechanism in Which Anchoring at the Nuclear Periphery Precedes Heterochromatin Formation10.3390/cells101128102073-4409https://doaj.org/article/fc8e847ee24e4172ae27a15b8d00532b2021-10-01T00:00:00Zhttps://www.mdpi.com/2073-4409/10/11/2810https://doaj.org/toc/2073-4409In eukaryotic cells, silent chromatin is mainly found at the nuclear periphery forming subnuclear compartments that favor silencing establishment. Here, we set up an inducible system to monitor silencing establishment at an ectopic locus in relation with its subnuclear localization in budding yeast. We previously showed that introducing LacI bound <i>lacO</i> arrays in proximity to gene flanked by <i>HML</i> silencers favors the recruitment of the yeast silencing complex SIR at this locus, leading to its silencing and anchoring at the nuclear periphery. Using an inducible version of this system, we show that silencing establishment is a stepwise process occurring over several cell cycles, with the progressive recruitment of the SIR complex. In contrast, we observed a rapid, SIR-independent perinuclear anchoring, induced by the high amount of LacI binding at the <i>lacO</i> array leading to nucleosome eviction at this array and to the phosphorylation of H2A in the neighboring nucleosomes by Mec1 kinase. While the initial phosphorylation of H2A (H2A-P) and perinuclear anchoring are independent of the SIR complex, its latter recruitment stabilizes H2A-P and reinforces the perinuclear anchoring. Finally, we showed that Sir3 spreading stabilizes nucleosomes and limits the access of specific DNA-binding protein to DNA.Isabelle LoïodiceMickael GarnierIvaylo NikolovAngela TaddeiMDPI AGarticlechromatinheterochromatinnuclear organizationgene regulationgene silencingFROSBiology (General)QH301-705.5ENCells, Vol 10, Iss 2810, p 2810 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	chromatin heterochromatin nuclear organization gene regulation gene silencing FROS Biology (General) QH301-705.5
spellingShingle	chromatin heterochromatin nuclear organization gene regulation gene silencing FROS Biology (General) QH301-705.5 Isabelle Loïodice Mickael Garnier Ivaylo Nikolov Angela Taddei An Inducible System for Silencing Establishment Reveals a Stepwise Mechanism in Which Anchoring at the Nuclear Periphery Precedes Heterochromatin Formation
description	In eukaryotic cells, silent chromatin is mainly found at the nuclear periphery forming subnuclear compartments that favor silencing establishment. Here, we set up an inducible system to monitor silencing establishment at an ectopic locus in relation with its subnuclear localization in budding yeast. We previously showed that introducing LacI bound <i>lacO</i> arrays in proximity to gene flanked by <i>HML</i> silencers favors the recruitment of the yeast silencing complex SIR at this locus, leading to its silencing and anchoring at the nuclear periphery. Using an inducible version of this system, we show that silencing establishment is a stepwise process occurring over several cell cycles, with the progressive recruitment of the SIR complex. In contrast, we observed a rapid, SIR-independent perinuclear anchoring, induced by the high amount of LacI binding at the <i>lacO</i> array leading to nucleosome eviction at this array and to the phosphorylation of H2A in the neighboring nucleosomes by Mec1 kinase. While the initial phosphorylation of H2A (H2A-P) and perinuclear anchoring are independent of the SIR complex, its latter recruitment stabilizes H2A-P and reinforces the perinuclear anchoring. Finally, we showed that Sir3 spreading stabilizes nucleosomes and limits the access of specific DNA-binding protein to DNA.
format	article
author	Isabelle Loïodice Mickael Garnier Ivaylo Nikolov Angela Taddei
author_facet	Isabelle Loïodice Mickael Garnier Ivaylo Nikolov Angela Taddei
author_sort	Isabelle Loïodice
title	An Inducible System for Silencing Establishment Reveals a Stepwise Mechanism in Which Anchoring at the Nuclear Periphery Precedes Heterochromatin Formation
title_short	An Inducible System for Silencing Establishment Reveals a Stepwise Mechanism in Which Anchoring at the Nuclear Periphery Precedes Heterochromatin Formation
title_full	An Inducible System for Silencing Establishment Reveals a Stepwise Mechanism in Which Anchoring at the Nuclear Periphery Precedes Heterochromatin Formation
title_fullStr	An Inducible System for Silencing Establishment Reveals a Stepwise Mechanism in Which Anchoring at the Nuclear Periphery Precedes Heterochromatin Formation
title_full_unstemmed	An Inducible System for Silencing Establishment Reveals a Stepwise Mechanism in Which Anchoring at the Nuclear Periphery Precedes Heterochromatin Formation
title_sort	inducible system for silencing establishment reveals a stepwise mechanism in which anchoring at the nuclear periphery precedes heterochromatin formation
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/fc8e847ee24e4172ae27a15b8d00532b
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An Inducible System for Silencing Establishment Reveals a Stepwise Mechanism in Which Anchoring at the Nuclear Periphery Precedes Heterochromatin Formation

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