5ITH

TIA-1 RRM2 recognition of target oligonucleotide

Summary for 5ITH

• Entry DOI: 10.2210/pdb5ith/pdb
• Descriptor: Nucleolysin TIA-1 isoform p40, DNA (5'-D(*AP*CP*TP*CP*C*TP*TP*TP*TP*T)-3') (3 entities in total)
• Functional Keywords: 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
• Biological source: Homo sapiens (Human); More
• Cellular location: Cytoplasmic granule : P31483
• Total number of polymer chains: 2
• Total formula weight: 13209.46

Authors

Waris, S.,Wilce, J.A.,Wilce, M.C. (deposition date: 2016-03-16, release date: 2017-02-22, Last modification date: 2023-09-27)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (2.31 Å)