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The high kinetic stability of a G-quadruplex limits hnRNP F qRRM3 binding to G-tract RNA
The RNA binding protein heterogeneous nuclear ribonucleoprotein (hnRNP) F is involved in telomeres maintenance and pre-mRNA processing, such as alternative splicing and polyadenylation. It specifically recognizes RNA containing three consecutive guanines (G-tracts) that have the potential to assembl...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3575826/ https://www.ncbi.nlm.nih.gov/pubmed/23275549 http://dx.doi.org/10.1093/nar/gks1289 |
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author | Samatanga, Brighton Dominguez, Cyril Jelesarov, Ilian Allain, Frédéric H.-T. |
author_facet | Samatanga, Brighton Dominguez, Cyril Jelesarov, Ilian Allain, Frédéric H.-T. |
author_sort | Samatanga, Brighton |
collection | PubMed |
description | The RNA binding protein heterogeneous nuclear ribonucleoprotein (hnRNP) F is involved in telomeres maintenance and pre-mRNA processing, such as alternative splicing and polyadenylation. It specifically recognizes RNA containing three consecutive guanines (G-tracts) that have the potential to assemble into G-quadruplexes. We have proposed recently that hnRNP F could regulate alternative splicing by remodeling RNA structures, such as G-quadruplexes. However, the exact mechanism of hnRNP F binding to such RNA sequences remains unknown. Here, we have studied the binding of the third RNA binding domain of hnRNP F [quasi-RNA recognition motif 3 (qRRM3)] to G-tract RNA using isothermal titration calorimetry, circular dichroism and nuclear magnetic resonance spectroscopy. Our results show that qRRM3 binds specifically exclusively to single-stranded G-tracts (ssRNA), in contrast to previous reports stating that the G-quadruplex was recognized as well. Furthermore, we demonstrate that the pre-existent ssRNA/G-quadruplex equilibrium slows down the formation of the protein–ssRNA complex. Based on in vitro transcription assays, we show that the rate of the protein–RNA complex formation is faster than that of the G-quadruplex. We propose a model according to which hnRNP F could bind RNA co-transcriptionally and prevents G-quadruplex formation. |
format | Online Article Text |
id | pubmed-3575826 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-35758262013-02-19 The high kinetic stability of a G-quadruplex limits hnRNP F qRRM3 binding to G-tract RNA Samatanga, Brighton Dominguez, Cyril Jelesarov, Ilian Allain, Frédéric H.-T. Nucleic Acids Res RNA The RNA binding protein heterogeneous nuclear ribonucleoprotein (hnRNP) F is involved in telomeres maintenance and pre-mRNA processing, such as alternative splicing and polyadenylation. It specifically recognizes RNA containing three consecutive guanines (G-tracts) that have the potential to assemble into G-quadruplexes. We have proposed recently that hnRNP F could regulate alternative splicing by remodeling RNA structures, such as G-quadruplexes. However, the exact mechanism of hnRNP F binding to such RNA sequences remains unknown. Here, we have studied the binding of the third RNA binding domain of hnRNP F [quasi-RNA recognition motif 3 (qRRM3)] to G-tract RNA using isothermal titration calorimetry, circular dichroism and nuclear magnetic resonance spectroscopy. Our results show that qRRM3 binds specifically exclusively to single-stranded G-tracts (ssRNA), in contrast to previous reports stating that the G-quadruplex was recognized as well. Furthermore, we demonstrate that the pre-existent ssRNA/G-quadruplex equilibrium slows down the formation of the protein–ssRNA complex. Based on in vitro transcription assays, we show that the rate of the protein–RNA complex formation is faster than that of the G-quadruplex. We propose a model according to which hnRNP F could bind RNA co-transcriptionally and prevents G-quadruplex formation. Oxford University Press 2013-02 2012-12-25 /pmc/articles/PMC3575826/ /pubmed/23275549 http://dx.doi.org/10.1093/nar/gks1289 Text en © The Author(s) 2012. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/3.0/), which permits non-commercial reuse, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com. |
spellingShingle | RNA Samatanga, Brighton Dominguez, Cyril Jelesarov, Ilian Allain, Frédéric H.-T. The high kinetic stability of a G-quadruplex limits hnRNP F qRRM3 binding to G-tract RNA |
title | The high kinetic stability of a G-quadruplex limits hnRNP F qRRM3 binding to G-tract RNA |
title_full | The high kinetic stability of a G-quadruplex limits hnRNP F qRRM3 binding to G-tract RNA |
title_fullStr | The high kinetic stability of a G-quadruplex limits hnRNP F qRRM3 binding to G-tract RNA |
title_full_unstemmed | The high kinetic stability of a G-quadruplex limits hnRNP F qRRM3 binding to G-tract RNA |
title_short | The high kinetic stability of a G-quadruplex limits hnRNP F qRRM3 binding to G-tract RNA |
title_sort | high kinetic stability of a g-quadruplex limits hnrnp f qrrm3 binding to g-tract rna |
topic | RNA |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3575826/ https://www.ncbi.nlm.nih.gov/pubmed/23275549 http://dx.doi.org/10.1093/nar/gks1289 |
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