5ITH

TIA-1 RRM2 recognition of target oligonucleotide

5ITH の概要

• エントリーDOI: 10.2210/pdb5ith/pdb
• 分子名称: Nucleolysin TIA-1 isoform p40, DNA (5'-D(*AP*CP*TP*CP*C*TP*TP*TP*TP*T)-3') (3 entities in total)
• 機能のキーワード: rrm, rna/dna binding domain, poly u binding preference, aromatic base stacking interactions, dna binding protein-dna complex, rna binding protein-dna complex, rna binding protein/dna
• 由来する生物種: Homo sapiens (Human); 詳細
• 細胞内の位置: Cytoplasmic granule : P31483
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 13209.46

構造登録者

Waris, S.,Wilce, J.A.,Wilce, M.C. (登録日: 2016-03-16, 公開日: 2017-02-22, 最終更新日: 2023-09-27)

主引用文献

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.
TIA-1 RRM23 binding and recognition of target oligonucleotides.
Nucleic Acids Res., 45:4944-4957, 2017

PubMed 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 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.

PubMed: 28184449
DOI: 10.1093/nar/gkx102

実験手法

X-RAY DIFFRACTION (2.31 Å)