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1.
RNA Biol ; 12(12): 1289-300, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26512911

RESUMEN

We describe here a platform for high-throughput protein expression and interaction analysis aimed at identifying the RNA-interacting domainome. This approach combines the selection of a phage library displaying "filtered" open reading frames with next-generation DNA sequencing. The method was validated using an RNA bait corresponding to the AU-rich element of α-prothymosin, an RNA motif that promotes mRNA stability and translation through its interaction with the RNA-binding protein ELAVL1. With this strategy, we not only confirmed known RNA-binding proteins that specifically interact with the target RNA (such as ELAVL1/HuR and RBM38) but also identified proteins not previously known to be ARE-binding (R3HDM2 and RALY). We propose this technology as a novel approach for studying the RNA-binding proteome.


Asunto(s)
Elementos Ricos en Adenilato y Uridilato/genética , Sistemas de Lectura Abierta/genética , Dominios y Motivos de Interacción de Proteínas/genética , Precursores de Proteínas/genética , Proteínas de Unión al ARN/metabolismo , Timosina/análogos & derivados , Células HEK293 , Humanos , Unión Proteica , Precursores de Proteínas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reproducibilidad de los Resultados , Timosina/genética , Timosina/metabolismo
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