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Sci Rep ; 6: 31313, 2016 08 11.
Artículo en Inglés | MEDLINE | ID: mdl-27510448

RESUMEN

Circular RNAs (circRNAs) constitute a new class of noncoding RNAs in higher eukaryotes generated from pre-mRNAs by alternative splicing. Here we investigated in mammalian cells the association of circRNAs with proteins. Using glycerol gradient centrifugation, we characterized in cell lysates circRNA-protein complexes (circRNPs) of distinct sizes. By polysome-gradient fractionation we found no evidence for efficient translation of a set of abundant circRNAs in HeLa cells. To identify circRNPs with a specific protein component, we focused on IMP3 (IGF2BP3, insulin-like growth factor 2 binding protein 3), a known tumor marker and RNA-binding protein. Combining RNA-seq analysis of IMP3-co-immunoprecipitated RNA and filtering for circular-junction reads identified a set of IMP3-associated circRNAs, which were validated and characterized. In sum, our data suggest that specific circRNP families exist defined by a common protein component. In addition, this provides a general approach to identify circRNPs with a given protein component.


Asunto(s)
Proteínas de Unión al ARN/metabolismo , ARN/metabolismo , Análisis de Secuencia de ARN/métodos , Biología Computacional , Células HeLa , Humanos , Inmunoprecipitación , Unión Proteica , ARN/genética , ARN Circular
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