Perturbation of Chromatin Structure Globally Affects Localization and Recruitment of Splicing Factors

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Chromatin structure is an important factor in the functional coupling between transcription and mRNA processing, not only by regulating alternative splicing events, but also by contributing to exon recognition during constitutive splicing. We observed that depolarization of neuroblastoma cell membra...

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Autores principales:	Schor, I.E., Llères, D., Risso, G.J., Pawellek, A., Ule, J., Lamond, A.I., Kornblihtt, A.R.
Formato:	Artículo publishedVersion
Lenguaje:	Inglés
Publicado:	2012
Materias:	enhanced green fluorescent protein heterochromatin protein 1 heterochromatin protein 1alpha histone H3 histone H4 messenger RNA nuclear protein protein SRSF1 protein SRSF2 trichostatin A unclassified drug untranslated RNA article bioaccumulation cell nucleus cellular distribution chromatin structure controlled study gene control gene function histone acetylation human human cell MALAT1 gene molecular imaging molecular model NCAM gene Neat1 gene protein determination protein function protein RNA binding regulatory mechanism RNA analysis RNA gene RNA splicing spliceosome structure analysis Acetylation Alternative Splicing Animals Cell Line Cell Nucleus Chromatin Histones Humans Hydroxamic Acids Membrane Potentials Models, Biological Protein Binding Protein Transport Ribonucleoproteins RNA Precursors RNA Splice Sites RNA Splicing RNA, Long Untranslated
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_19326203_v7_n11_p_Schor
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paperaa:paper_19326203_v7_n11_p_Schor
record_format	dspace
spelling	paperaa:paper_19326203_v7_n11_p_Schor2023-06-12T16:51:34Z Perturbation of Chromatin Structure Globally Affects Localization and Recruitment of Splicing Factors PLoS ONE 2012;7(11) Schor, I.E. Llères, D. Risso, G.J. Pawellek, A. Ule, J. Lamond, A.I. Kornblihtt, A.R. enhanced green fluorescent protein heterochromatin protein 1 heterochromatin protein 1alpha histone H3 histone H4 messenger RNA nuclear protein protein SRSF1 protein SRSF2 trichostatin A unclassified drug untranslated RNA article bioaccumulation cell nucleus cellular distribution chromatin structure controlled study gene control gene function histone acetylation human human cell MALAT1 gene molecular imaging molecular model NCAM gene Neat1 gene protein determination protein function protein RNA binding regulatory mechanism RNA analysis RNA gene RNA splicing spliceosome structure analysis Acetylation Alternative Splicing Animals Cell Line Cell Nucleus Chromatin Histones Humans Hydroxamic Acids Membrane Potentials Models, Biological Protein Binding Protein Transport Ribonucleoproteins RNA Precursors RNA Splice Sites RNA Splicing RNA, Long Untranslated Chromatin structure is an important factor in the functional coupling between transcription and mRNA processing, not only by regulating alternative splicing events, but also by contributing to exon recognition during constitutive splicing. We observed that depolarization of neuroblastoma cell membrane potential, which triggers general histone acetylation and regulates alternative splicing, causes a concentration of SR proteins in nuclear speckles. This prompted us to analyze the effect of chromatin structure on splicing factor distribution and dynamics. Here, we show that induction of histone hyper-acetylation results in the accumulation in speckles of multiple splicing factors in different cell types. In addition, a similar effect is observed after depletion of the heterochromatic protein HP1α, associated with repressive chromatin. We used advanced imaging approaches to analyze in detail both the structural organization of the speckle compartment and nuclear distribution of splicing factors, as well as studying direct interactions between splicing factors and their association with chromatin in vivo. The results support a model where perturbation of normal chromatin structure decreases the recruitment efficiency of splicing factors to nascent RNAs, thus causing their accumulation in speckles, which buffer the amount of free molecules in the nucleoplasm. To test this, we analyzed the recruitment of the general splicing factor U2AF65 to nascent RNAs by iCLIP technique, as a way to monitor early spliceosome assembly. We demonstrate that indeed histone hyper-acetylation decreases recruitment of U2AF65 to bulk 3′ splice sites, coincident with the change in its localization. In addition, prior to the maximum accumulation in speckles, ~20% of genes already show a tendency to decreased binding, while U2AF65 seems to increase its binding to the speckle-located ncRNA MALAT1. All together, the combined imaging and biochemical approaches support a model where chromatin structure is essential for efficient co-transcriptional recruitment of general and regulatory splicing factors to pre-mRNA. © 2012 Schor et al. Fil:Schor, I.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Risso, G.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Kornblihtt, A.R. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2012 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_19326203_v7_n11_p_Schor
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language	Inglés
orig_language_str_mv	eng
topic	enhanced green fluorescent protein heterochromatin protein 1 heterochromatin protein 1alpha histone H3 histone H4 messenger RNA nuclear protein protein SRSF1 protein SRSF2 trichostatin A unclassified drug untranslated RNA article bioaccumulation cell nucleus cellular distribution chromatin structure controlled study gene control gene function histone acetylation human human cell MALAT1 gene molecular imaging molecular model NCAM gene Neat1 gene protein determination protein function protein RNA binding regulatory mechanism RNA analysis RNA gene RNA splicing spliceosome structure analysis Acetylation Alternative Splicing Animals Cell Line Cell Nucleus Chromatin Histones Humans Hydroxamic Acids Membrane Potentials Models, Biological Protein Binding Protein Transport Ribonucleoproteins RNA Precursors RNA Splice Sites RNA Splicing RNA, Long Untranslated
spellingShingle	enhanced green fluorescent protein heterochromatin protein 1 heterochromatin protein 1alpha histone H3 histone H4 messenger RNA nuclear protein protein SRSF1 protein SRSF2 trichostatin A unclassified drug untranslated RNA article bioaccumulation cell nucleus cellular distribution chromatin structure controlled study gene control gene function histone acetylation human human cell MALAT1 gene molecular imaging molecular model NCAM gene Neat1 gene protein determination protein function protein RNA binding regulatory mechanism RNA analysis RNA gene RNA splicing spliceosome structure analysis Acetylation Alternative Splicing Animals Cell Line Cell Nucleus Chromatin Histones Humans Hydroxamic Acids Membrane Potentials Models, Biological Protein Binding Protein Transport Ribonucleoproteins RNA Precursors RNA Splice Sites RNA Splicing RNA, Long Untranslated Schor, I.E. Llères, D. Risso, G.J. Pawellek, A. Ule, J. Lamond, A.I. Kornblihtt, A.R. Perturbation of Chromatin Structure Globally Affects Localization and Recruitment of Splicing Factors
topic_facet	enhanced green fluorescent protein heterochromatin protein 1 heterochromatin protein 1alpha histone H3 histone H4 messenger RNA nuclear protein protein SRSF1 protein SRSF2 trichostatin A unclassified drug untranslated RNA article bioaccumulation cell nucleus cellular distribution chromatin structure controlled study gene control gene function histone acetylation human human cell MALAT1 gene molecular imaging molecular model NCAM gene Neat1 gene protein determination protein function protein RNA binding regulatory mechanism RNA analysis RNA gene RNA splicing spliceosome structure analysis Acetylation Alternative Splicing Animals Cell Line Cell Nucleus Chromatin Histones Humans Hydroxamic Acids Membrane Potentials Models, Biological Protein Binding Protein Transport Ribonucleoproteins RNA Precursors RNA Splice Sites RNA Splicing RNA, Long Untranslated
description	Chromatin structure is an important factor in the functional coupling between transcription and mRNA processing, not only by regulating alternative splicing events, but also by contributing to exon recognition during constitutive splicing. We observed that depolarization of neuroblastoma cell membrane potential, which triggers general histone acetylation and regulates alternative splicing, causes a concentration of SR proteins in nuclear speckles. This prompted us to analyze the effect of chromatin structure on splicing factor distribution and dynamics. Here, we show that induction of histone hyper-acetylation results in the accumulation in speckles of multiple splicing factors in different cell types. In addition, a similar effect is observed after depletion of the heterochromatic protein HP1α, associated with repressive chromatin. We used advanced imaging approaches to analyze in detail both the structural organization of the speckle compartment and nuclear distribution of splicing factors, as well as studying direct interactions between splicing factors and their association with chromatin in vivo. The results support a model where perturbation of normal chromatin structure decreases the recruitment efficiency of splicing factors to nascent RNAs, thus causing their accumulation in speckles, which buffer the amount of free molecules in the nucleoplasm. To test this, we analyzed the recruitment of the general splicing factor U2AF65 to nascent RNAs by iCLIP technique, as a way to monitor early spliceosome assembly. We demonstrate that indeed histone hyper-acetylation decreases recruitment of U2AF65 to bulk 3′ splice sites, coincident with the change in its localization. In addition, prior to the maximum accumulation in speckles, ~20% of genes already show a tendency to decreased binding, while U2AF65 seems to increase its binding to the speckle-located ncRNA MALAT1. All together, the combined imaging and biochemical approaches support a model where chromatin structure is essential for efficient co-transcriptional recruitment of general and regulatory splicing factors to pre-mRNA. © 2012 Schor et al.
format	Artículo Artículo publishedVersion
author	Schor, I.E. Llères, D. Risso, G.J. Pawellek, A. Ule, J. Lamond, A.I. Kornblihtt, A.R.
author_facet	Schor, I.E. Llères, D. Risso, G.J. Pawellek, A. Ule, J. Lamond, A.I. Kornblihtt, A.R.
author_sort	Schor, I.E.
title	Perturbation of Chromatin Structure Globally Affects Localization and Recruitment of Splicing Factors
title_short	Perturbation of Chromatin Structure Globally Affects Localization and Recruitment of Splicing Factors
title_full	Perturbation of Chromatin Structure Globally Affects Localization and Recruitment of Splicing Factors
title_fullStr	Perturbation of Chromatin Structure Globally Affects Localization and Recruitment of Splicing Factors
title_full_unstemmed	Perturbation of Chromatin Structure Globally Affects Localization and Recruitment of Splicing Factors
title_sort	perturbation of chromatin structure globally affects localization and recruitment of splicing factors
publishDate	2012
url	http://hdl.handle.net/20.500.12110/paper_19326203_v7_n11_p_Schor
work_keys_str_mv	AT schorie perturbationofchromatinstructuregloballyaffectslocalizationandrecruitmentofsplicingfactors AT lleresd perturbationofchromatinstructuregloballyaffectslocalizationandrecruitmentofsplicingfactors AT rissogj perturbationofchromatinstructuregloballyaffectslocalizationandrecruitmentofsplicingfactors AT pawelleka perturbationofchromatinstructuregloballyaffectslocalizationandrecruitmentofsplicingfactors AT ulej perturbationofchromatinstructuregloballyaffectslocalizationandrecruitmentofsplicingfactors AT lamondai perturbationofchromatinstructuregloballyaffectslocalizationandrecruitmentofsplicingfactors AT kornblihttar perturbationofchromatinstructuregloballyaffectslocalizationandrecruitmentofsplicingfactors
_version_	1769810354265653248

Perturbation of Chromatin Structure Globally Affects Localization and Recruitment of Splicing Factors

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