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Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation

TDP-43 is a nuclear protein involved in many aspects of RNA metabolism. To ensure cellular viability, its expression levels within cells must be tightly regulated. We have previously demonstrated that TDP-43 autoregulation occurs through the activation of a normally silent intron in its 3′-UTR seque...

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Autores principales: Bembich, Sara, Herzog, Jeremias S., De Conti, Laura, Stuani, Cristiana, Avendaño-Vázquez, S. Eréndira, Buratti, Emanuele, Baralle, Marco, Baralle, Francisco E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2014
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3950720/
https://www.ncbi.nlm.nih.gov/pubmed/24369426
http://dx.doi.org/10.1093/nar/gkt1343
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author Bembich, Sara
Herzog, Jeremias S.
De Conti, Laura
Stuani, Cristiana
Avendaño-Vázquez, S. Eréndira
Buratti, Emanuele
Baralle, Marco
Baralle, Francisco E.
author_facet Bembich, Sara
Herzog, Jeremias S.
De Conti, Laura
Stuani, Cristiana
Avendaño-Vázquez, S. Eréndira
Buratti, Emanuele
Baralle, Marco
Baralle, Francisco E.
author_sort Bembich, Sara
collection PubMed
description TDP-43 is a nuclear protein involved in many aspects of RNA metabolism. To ensure cellular viability, its expression levels within cells must be tightly regulated. We have previously demonstrated that TDP-43 autoregulation occurs through the activation of a normally silent intron in its 3′-UTR sequence that results in the use of alternative polyadenylation sites. In this work, we analyse which is the dominant event in autoregulation: the recognition of the splice sites of 3′-UTR intron 7 or the intrinsic quality of the alternative polyadenylation sites. A panel of minigene constructs was tested for autoregulation functionality, protein production and subcellular messenger RNA localization. Our data clearly indicate that constitutive spliceosome complex formation across intron 7 does not lead to high protein production but, on the contrary, to lower TDP-43 messenger RNA and protein levels. This is due to altered nucleocytoplasmic distribution of the RNA that is mostly retained in the nucleus and degraded. This study provides a novel in-depth characterization of how RNA binding proteins can autoregulate their own levels within cells, an essential regulatory process in maintaining cellular viability.
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spelling pubmed-39507202014-03-12 Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation Bembich, Sara Herzog, Jeremias S. De Conti, Laura Stuani, Cristiana Avendaño-Vázquez, S. Eréndira Buratti, Emanuele Baralle, Marco Baralle, Francisco E. Nucleic Acids Res TDP-43 is a nuclear protein involved in many aspects of RNA metabolism. To ensure cellular viability, its expression levels within cells must be tightly regulated. We have previously demonstrated that TDP-43 autoregulation occurs through the activation of a normally silent intron in its 3′-UTR sequence that results in the use of alternative polyadenylation sites. In this work, we analyse which is the dominant event in autoregulation: the recognition of the splice sites of 3′-UTR intron 7 or the intrinsic quality of the alternative polyadenylation sites. A panel of minigene constructs was tested for autoregulation functionality, protein production and subcellular messenger RNA localization. Our data clearly indicate that constitutive spliceosome complex formation across intron 7 does not lead to high protein production but, on the contrary, to lower TDP-43 messenger RNA and protein levels. This is due to altered nucleocytoplasmic distribution of the RNA that is mostly retained in the nucleus and degraded. This study provides a novel in-depth characterization of how RNA binding proteins can autoregulate their own levels within cells, an essential regulatory process in maintaining cellular viability. Oxford University Press 2014-03 2013-12-24 /pmc/articles/PMC3950720/ /pubmed/24369426 http://dx.doi.org/10.1093/nar/gkt1343 Text en © The Author(s) 2013. Published by Oxford University Press. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Bembich, Sara
Herzog, Jeremias S.
De Conti, Laura
Stuani, Cristiana
Avendaño-Vázquez, S. Eréndira
Buratti, Emanuele
Baralle, Marco
Baralle, Francisco E.
Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation
title Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation
title_full Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation
title_fullStr Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation
title_full_unstemmed Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation
title_short Predominance of spliceosomal complex formation over polyadenylation site selection in TDP-43 autoregulation
title_sort predominance of spliceosomal complex formation over polyadenylation site selection in tdp-43 autoregulation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3950720/
https://www.ncbi.nlm.nih.gov/pubmed/24369426
http://dx.doi.org/10.1093/nar/gkt1343
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