Journal
NUCLEIC ACIDS RESEARCH
Volume 45, Issue 8, Pages 4944-4957Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkx102
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Funding
- National Health and Medical Research Council [APP1105801]
- National Health and Medical Research Senior Research Fellowship [FP1079611]
- NIA-IRP, NIH
- Andalusian Government [P11-CVI-7216, BIO198]
- Spanish Ministry of Science and Innovation [BFU2015-71017-P]
- Monash University
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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 angstrom 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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