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.
|Titolo:||Identification of novel proteins binding the AU-rich element of α-prothymosin mRNA through the selection of open reading frames (RIDome)|
|Autori:||Patrucco, L; Peano, C; Chiesa, A; Guida, F; Luisi, I; Boria, I; Mignone, F; De Bellis, G; Zucchelli, S; Gustincich, S; Santoro, C; Sblattero, D; Cotella, D|
|Data di pubblicazione:||2015|
|Digital Object Identifier (DOI):||10.1080/15476286.2015.1107702|
|Appare nelle tipologie:||1.1 Journal article|