2024-03-28T08:53:45Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1500242021-12-28T15:57:48Zcom_10261_38com_10261_5col_10261_291
00925njm 22002777a 4500
dc
Waris, Saboora
author
García-Mauriño, Sofía M.
author
Sivakumaran, Andrew
author
Beckham, Simone A.
author
Loughlin, Fionna E.
author
Gorospe, Myriam
author
Díaz-Moreno, Irene
author
Wilce, Matthew C.J.
author
Wilce, Jacqueline A.
author
2017
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.
Nucleic Acids Research, 45(8):4944-4957 (2017)
http://hdl.handle.net/10261/150024
10.1093/nar/gkx102
28184449
TIA-1 RRM23 binding and recognition of target oligonucleotides