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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2017
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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. |
format | Online Article Text |
id | pubmed-5416816 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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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