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.

Identification of novel proteins binding the AU-rich element of α-prothymosin mRNA through the selection of open reading frames (RIDome) / Patrucco, L; Peano, C; Chiesa, A; Guida, Filomena; Luisi, I; Boria, I; Mignone, F; De Bellis, G; Zucchelli, Silvia; Gustincich, Stefano; Santoro, C; Sblattero, D; Cotella, D.. - In: RNA BIOLOGY. - ISSN 1547-6286. - 12:12(2015), pp. 1289-1300. [10.1080/15476286.2015.1107702]

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

Guida, Filomena;Zucchelli, Silvia;Gustincich, Stefano;
2015-01-01

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 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.
2015
12
12
1289
1300
https://www.ncbi.nlm.nih.gov/pubmed/?term=Identification+of+novel+proteins+binding+the+AU-rich+element+of+%CE%B1-prothymosin+mRNA+through+the+selection+of+open
Patrucco, L; Peano, C; Chiesa, A; Guida, Filomena; Luisi, I; Boria, I; Mignone, F; De Bellis, G; Zucchelli, Silvia; Gustincich, Stefano; Santoro, C; S...espandi
File in questo prodotto:
File Dimensione Formato  
Patrucco 2015 RNA Biol.pdf

non disponibili

Descrizione: postprint
Tipologia: Documento in Post-print
Licenza: Non specificato
Dimensione 3.05 MB
Formato Adobe PDF
3.05 MB Adobe PDF   Visualizza/Apri   Richiedi una copia

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.11767/14402
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 4
  • ???jsp.display-item.citation.isi??? 5
social impact