RESUMEN

The human prototypical SR protein SRSF1 is an oncoprotein that contains two RRMs and plays a pivotal role in RNA metabolism. We determined the structure of the RRM1 bound to RNA and found that the domain binds preferentially to a CN motif (N is for any nucleotide). Based on this solution structure, we engineered a protein containing a single glutamate to asparagine mutation (E87N), which gains the ability to bind to uridines and thereby activates SMN exon7 inclusion, a strategy that is used to cure spinal muscular atrophy. Finally, we revealed that the flexible inter-RRM linker of SRSF1 allows RRM1 to bind RNA on both sides of RRM2 binding site. Besides revealing an unexpected bimodal mode of interaction of SRSF1 with RNA, which will be of interest to design new therapeutic strategies, this study brings a new perspective on the mode of action of SRSF1 in cells.

Asunto(s)

Motivo de Reconocimiento de ARN/genética , Sitios de Empalme de ARN/genética , Empalme del ARN , Factores de Empalme Serina-Arginina/metabolismo , Proteína 1 para la Supervivencia de la Neurona Motora/genética , Sustitución de Aminoácidos , Asparagina/genética , Biología Computacional , Exones/genética , Ácido Glutámico/genética , Células HEK293 , Humanos , Simulación de Dinámica Molecular , Atrofia Muscular Espinal/genética , Atrofia Muscular Espinal/terapia , Resonancia Magnética Nuclear Biomolecular , Ingeniería de Proteínas , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/ultraestructura , Factores de Empalme Serina-Arginina/genética , Factores de Empalme Serina-Arginina/aislamiento & purificación , Factores de Empalme Serina-Arginina/ultraestructura , Uridina/metabolismo