Abstract：
:Protein-RNA networks are ubiquitous and central in biological control. We present an approach termed RNA Tagging that enables the user to identify protein-RNA interactions in vivo by analyzing purified cellular RNA, without protein purification or cross-linking. An RNA-binding protein of interest is fused to an enzyme that adds uridines to the end of RNA. RNA targets bound by the chimeric protein in vivo are covalently marked with uridines and subsequently identified from extracted RNA via high-throughput sequencing. We used this approach to identify hundreds of RNAs bound by a Saccharomyces cerevisiae PUF protein, Puf3p. The results showed that although RNA-binding proteins productively bind specific RNAs to control their function, they also 'sample' RNAs without exerting a regulatory effect. We used the method to uncover hundreds of new and likely regulated targets for a protein without canonical RNA-binding domains, Bfr1p. RNA Tagging is well suited to detect and analyze protein-RNA networks in vivo.
journal_name
Nat Methodsjournal_title
Nature methodsauthors
Lapointe CP,Wilinski D,Saunders HA,Wickens Mdoi
10.1038/nmeth.3651subject
Has Abstractpub_date
2015-12-01 00:00:00pages
1163-70issue
12eissn
1548-7091issn
1548-7105pii
nmeth.3651journal_volume
12pub_type
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