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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...

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Autores principales: Samatanga, Brighton, Dominguez, Cyril, Jelesarov, Ilian, Allain, Frédéric H.-T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2013
Materias:
RNA
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.
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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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