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Barrier-to-Autointegration Factor Proteome Reveals Chromatin-Regulatory Partners

Nuclear lamin filaments and associated proteins form a nucleoskeletal (“lamina”) network required for transcription, replication, chromatin organization and epigenetic regulation in metazoans. Lamina defects cause human disease (“laminopathies”) and are linked to aging. Barrier-to-autointegration fa...

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Autores principales: Montes de Oca, Rocío, Shoemaker, Christopher J., Gucek, Marjan, Cole, Robert N., Wilson, Katherine L.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2739719/
https://www.ncbi.nlm.nih.gov/pubmed/19759913
http://dx.doi.org/10.1371/journal.pone.0007050
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author Montes de Oca, Rocío
Shoemaker, Christopher J.
Gucek, Marjan
Cole, Robert N.
Wilson, Katherine L.
author_facet Montes de Oca, Rocío
Shoemaker, Christopher J.
Gucek, Marjan
Cole, Robert N.
Wilson, Katherine L.
author_sort Montes de Oca, Rocío
collection PubMed
description Nuclear lamin filaments and associated proteins form a nucleoskeletal (“lamina”) network required for transcription, replication, chromatin organization and epigenetic regulation in metazoans. Lamina defects cause human disease (“laminopathies”) and are linked to aging. Barrier-to-autointegration factor (BAF) is a mobile and essential component of the nuclear lamina that binds directly to histones, lamins and LEM-domain proteins, including the inner nuclear membrane protein emerin, and has roles in chromatin structure, mitosis and gene regulation. To understand BAF's mechanisms of action, BAF associated proteins were affinity-purified from HeLa cell nuclear lysates using BAF-conjugated beads, and identified by tandem mass spectrometry or independently identified and quantified using the iTRAQ method. We recovered A- and B-type lamins and core histones, all known to bind BAF directly, plus four human transcription factors (Requiem, NonO, p15, LEDGF), disease-linked proteins (e.g., Huntingtin, Treacle) and several proteins and enzymes that regulate chromatin. Association with endogenous BAF was independently validated by co-immunoprecipitation from HeLa cells for seven candidates including Requiem, poly(ADP-ribose) polymerase 1 (PARP1), retinoblastoma binding protein 4 (RBBP4), damage-specific DNA binding protein 1 (DDB1) and DDB2. Interestingly, endogenous BAF and emerin each associated with DDB2 and CUL4A in a UV- and time-dependent manner, suggesting BAF and emerin have dynamic roles in genome integrity and might help couple DNA damage responses to the nuclear lamina network. We conclude this proteome is a rich source of candidate partners for BAF and potentially also A- and B-type lamins, which may reveal how chromatin regulation and genome integrity are linked to nuclear structure.
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spelling pubmed-27397192009-09-17 Barrier-to-Autointegration Factor Proteome Reveals Chromatin-Regulatory Partners Montes de Oca, Rocío Shoemaker, Christopher J. Gucek, Marjan Cole, Robert N. Wilson, Katherine L. PLoS One Research Article Nuclear lamin filaments and associated proteins form a nucleoskeletal (“lamina”) network required for transcription, replication, chromatin organization and epigenetic regulation in metazoans. Lamina defects cause human disease (“laminopathies”) and are linked to aging. Barrier-to-autointegration factor (BAF) is a mobile and essential component of the nuclear lamina that binds directly to histones, lamins and LEM-domain proteins, including the inner nuclear membrane protein emerin, and has roles in chromatin structure, mitosis and gene regulation. To understand BAF's mechanisms of action, BAF associated proteins were affinity-purified from HeLa cell nuclear lysates using BAF-conjugated beads, and identified by tandem mass spectrometry or independently identified and quantified using the iTRAQ method. We recovered A- and B-type lamins and core histones, all known to bind BAF directly, plus four human transcription factors (Requiem, NonO, p15, LEDGF), disease-linked proteins (e.g., Huntingtin, Treacle) and several proteins and enzymes that regulate chromatin. Association with endogenous BAF was independently validated by co-immunoprecipitation from HeLa cells for seven candidates including Requiem, poly(ADP-ribose) polymerase 1 (PARP1), retinoblastoma binding protein 4 (RBBP4), damage-specific DNA binding protein 1 (DDB1) and DDB2. Interestingly, endogenous BAF and emerin each associated with DDB2 and CUL4A in a UV- and time-dependent manner, suggesting BAF and emerin have dynamic roles in genome integrity and might help couple DNA damage responses to the nuclear lamina network. We conclude this proteome is a rich source of candidate partners for BAF and potentially also A- and B-type lamins, which may reveal how chromatin regulation and genome integrity are linked to nuclear structure. Public Library of Science 2009-09-16 /pmc/articles/PMC2739719/ /pubmed/19759913 http://dx.doi.org/10.1371/journal.pone.0007050 Text en Montes de Oca et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Montes de Oca, Rocío
Shoemaker, Christopher J.
Gucek, Marjan
Cole, Robert N.
Wilson, Katherine L.
Barrier-to-Autointegration Factor Proteome Reveals Chromatin-Regulatory Partners
title Barrier-to-Autointegration Factor Proteome Reveals Chromatin-Regulatory Partners
title_full Barrier-to-Autointegration Factor Proteome Reveals Chromatin-Regulatory Partners
title_fullStr Barrier-to-Autointegration Factor Proteome Reveals Chromatin-Regulatory Partners
title_full_unstemmed Barrier-to-Autointegration Factor Proteome Reveals Chromatin-Regulatory Partners
title_short Barrier-to-Autointegration Factor Proteome Reveals Chromatin-Regulatory Partners
title_sort barrier-to-autointegration factor proteome reveals chromatin-regulatory partners
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2739719/
https://www.ncbi.nlm.nih.gov/pubmed/19759913
http://dx.doi.org/10.1371/journal.pone.0007050
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