Identification of novel proteins binding the AU-rich element of ?-prothymosin mRNA through the selection of open reading frames (RIDome).

ABSTRACT: 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 ?-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.

SUBMITTER: Patrucco L 

PROVIDER: S-EPMC4829324 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature