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TIA-1 RRM23 binding and recognition of target oligonucleotides

TIA-1 (T-cell restricted intracellular antigen-1) is an RNA-binding protein involved in splicing and translational repression. It mainly interacts with RNA via its second and third RNA recognition motifs (RRMs), with specificity for U-rich sequences directed by RRM2. It has recently been shown that...

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Autores principales: Waris, Saboora, García-Mauriño, Sofía M., Sivakumaran, Andrew, Beckham, Simone A., Loughlin, Fionna E., Gorospe, Myriam, Díaz-Moreno, Irene, Wilce, Matthew C.J., Wilce, Jacqueline A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5416816/
https://www.ncbi.nlm.nih.gov/pubmed/28184449
http://dx.doi.org/10.1093/nar/gkx102
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author Waris, Saboora
García-Mauriño, Sofía M.
Sivakumaran, Andrew
Beckham, Simone A.
Loughlin, Fionna E.
Gorospe, Myriam
Díaz-Moreno, Irene
Wilce, Matthew C.J.
Wilce, Jacqueline A.
author_facet Waris, Saboora
García-Mauriño, Sofía M.
Sivakumaran, Andrew
Beckham, Simone A.
Loughlin, Fionna E.
Gorospe, Myriam
Díaz-Moreno, Irene
Wilce, Matthew C.J.
Wilce, Jacqueline A.
author_sort Waris, Saboora
collection PubMed
description TIA-1 (T-cell restricted intracellular antigen-1) is an RNA-binding protein involved in splicing and translational repression. It mainly interacts with RNA via its second and third RNA recognition motifs (RRMs), with specificity for U-rich sequences directed by RRM2. It has recently been shown that RRM3 also contributes to binding, with preferential binding for C-rich sequences. Here we designed UC-rich and CU-rich 10-nt sequences for engagement of both RRM2 and RRM3 and demonstrated that the TIA-1 RRM23 construct preferentially binds the UC-rich RNA ligand (5΄-UUUUUACUCC-3΄). Interestingly, this binding depends on the presence of Lys274 that is C-terminal to RRM3 and binding to equivalent DNA sequences occurs with similar affinity. Small-angle X-ray scattering was used to demonstrate that, upon complex formation with target RNA or DNA, TIA-1 RRM23 adopts a compact structure, showing that both RRMs engage with the target 10-nt sequences to form the complex. We also report the crystal structure of TIA-1 RRM2 in complex with DNA to 2.3 Å resolution providing the first atomic resolution structure of any TIA protein RRM in complex with oligonucleotide. Together our data support a specific mode of TIA-1 RRM23 interaction with target oligonucleotides consistent with the role of TIA-1 in binding RNA to regulate gene expression.
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spelling pubmed-54168162017-05-05 TIA-1 RRM23 binding and recognition of target oligonucleotides Waris, Saboora García-Mauriño, Sofía M. Sivakumaran, Andrew Beckham, Simone A. Loughlin, Fionna E. Gorospe, Myriam Díaz-Moreno, Irene Wilce, Matthew C.J. Wilce, Jacqueline A. Nucleic Acids Res Structural Biology TIA-1 (T-cell restricted intracellular antigen-1) is an RNA-binding protein involved in splicing and translational repression. It mainly interacts with RNA via its second and third RNA recognition motifs (RRMs), with specificity for U-rich sequences directed by RRM2. It has recently been shown that RRM3 also contributes to binding, with preferential binding for C-rich sequences. Here we designed UC-rich and CU-rich 10-nt sequences for engagement of both RRM2 and RRM3 and demonstrated that the TIA-1 RRM23 construct preferentially binds the UC-rich RNA ligand (5΄-UUUUUACUCC-3΄). Interestingly, this binding depends on the presence of Lys274 that is C-terminal to RRM3 and binding to equivalent DNA sequences occurs with similar affinity. Small-angle X-ray scattering was used to demonstrate that, upon complex formation with target RNA or DNA, TIA-1 RRM23 adopts a compact structure, showing that both RRMs engage with the target 10-nt sequences to form the complex. We also report the crystal structure of TIA-1 RRM2 in complex with DNA to 2.3 Å resolution providing the first atomic resolution structure of any TIA protein RRM in complex with oligonucleotide. Together our data support a specific mode of TIA-1 RRM23 interaction with target oligonucleotides consistent with the role of TIA-1 in binding RNA to regulate gene expression. Oxford University Press 2017-05-05 2017-02-10 /pmc/articles/PMC5416816/ /pubmed/28184449 http://dx.doi.org/10.1093/nar/gkx102 Text en © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Structural Biology
Waris, Saboora
García-Mauriño, Sofía M.
Sivakumaran, Andrew
Beckham, Simone A.
Loughlin, Fionna E.
Gorospe, Myriam
Díaz-Moreno, Irene
Wilce, Matthew C.J.
Wilce, Jacqueline A.
TIA-1 RRM23 binding and recognition of target oligonucleotides
title TIA-1 RRM23 binding and recognition of target oligonucleotides
title_full TIA-1 RRM23 binding and recognition of target oligonucleotides
title_fullStr TIA-1 RRM23 binding and recognition of target oligonucleotides
title_full_unstemmed TIA-1 RRM23 binding and recognition of target oligonucleotides
title_short TIA-1 RRM23 binding and recognition of target oligonucleotides
title_sort tia-1 rrm23 binding and recognition of target oligonucleotides
topic Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5416816/
https://www.ncbi.nlm.nih.gov/pubmed/28184449
http://dx.doi.org/10.1093/nar/gkx102
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