TIA-1 RRM23 binding and recognition of target oligonucleotides

Saboora Waris, Sofia M. Garcia-Maurino, Andrew Sivakumaran, Simone A. Beckham, Fionna E. Loughlin, Myriam Gorospe, Irene Diaz-Moreno, Matthew C.J. Wilce, Jacqueline A. Wilce

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6 Citations (Scopus)

Abstract

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 CUrich 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 °A 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.

Original languageEnglish
Pages (from-to)4944-4957
Number of pages14
JournalNucleic Acids Research
Volume45
Issue number8
DOIs
Publication statusPublished - 5 May 2017

Keywords

  • transient ischemic attack
  • dna
  • oligonucleotides
  • rna

Cite this

Waris, S., Garcia-Maurino, S. M., Sivakumaran, A., Beckham, S. A., Loughlin, F. E., Gorospe, M., Diaz-Moreno, I., Wilce, M. C. J., & Wilce, J. A. (2017). TIA-1 RRM23 binding and recognition of target oligonucleotides. Nucleic Acids Research, 45(8), 4944-4957. https://doi.org/10.1093/nar/gkx102