Artículo
The binding of TIA-1 to RNA C-rich sequences is driven by its C-terminal RRM domain
Autor/es | Aroca Aguilar, Ángeles
Cruz Gallardo, Isabel Díaz Moreno, Irene Angulo Álvarez, Jesús Gunzburg, Menachen J. Sivakumaran, Andrew Yoon, Je-Hyun |
Departamento | Universidad de Sevilla. Departamento de Bioquímica Vegetal y Biología Molecular |
Fecha de publicación | 2014 |
Fecha de depósito | 2018-02-08 |
Publicado en |
|
Resumen | T-cell intracellular antigen-1 (TIA-1) is a key DNA/RNA binding protein that regulates translation by sequestering target mRNAs in stress granules (SG) in response to stress conditions. TIA-1 possesses three RNA recognition ... T-cell intracellular antigen-1 (TIA-1) is a key DNA/RNA binding protein that regulates translation by sequestering target mRNAs in stress granules (SG) in response to stress conditions. TIA-1 possesses three RNA recognition motifs (RRM) along with a glutamine-rich domain, with the central domains (RRM2 and RRM3) acting as RNA binding platforms. While the RRM2 domain, which displays high affinity for U-rich RNA sequences, is primarily responsible for interaction with RNA, the contribution of RRM3 to bind RNA as well as the target RNA sequences that it binds preferentially are still unknown. Here we combined nuclear magnetic resonance (NMR) and surface plasmon resonance (SPR) techniques to elucidate the sequence specificity of TIA-1 RRM3. With a novel approach using saturation transfer difference NMR (STD-NMR) to quantify protein-nucleic acids interactions, we demonstrate that isolated RRM3 binds to both C- and U-rich stretches with micromolar affinity. In combination with RRM2 and in the context of full-length TIA-1, RRM3 significantly enhanced the binding to RNA, particularly to cytosine-rich RNA oligos, as assessed by biotinylated RNA pull-down analysis. Our findings provide new insight into the role of RRM3 in regulating TIA-1 binding to C-rich stretches, that are abundant at the 5' TOPs (5' terminal oligopyrimidine tracts) of mRNAs whose translation is repressed under stress situations |
Agencias financiadoras | Junta de Andalucía |
Identificador del proyecto | P07-CVI-02896
P11-CVI-7216 BIO198 |
Cita | Aroca Aguilar, Á., Cruz Gallardo, I., Díaz Moreno, I., Angulo Álvarez, J., Gunzburg, M.J., Sivakumaran, A. y Yoon, J. (2014). The binding of TIA-1 to RNA C-rich sequences is driven by its C-terminal RRM domain. RNA Biology, 11 (6), 766-776. |
Ficheros | Tamaño | Formato | Ver | Descripción |
---|---|---|---|---|
csic302014RNABIOL0065R (2).pdf | 277.4Kb | [PDF] | Ver/ | |