Rapid and systematic analysis of the RNA recognition specificities of RNA-binding proteins.
Nat Biotechnol, 2009/7;27(7):667-70.
Ray D[1], Kazan H, Chan ET, Peña Castillo L, Chaudhry S, Talukder S, Blencowe BJ, Morris Q, Hughes TR
Affiliations
PMID: 19561594DOI: 10.1038/nbt.1550
Impact factor: 68.164
Abstract
Metazoan genomes encode hundreds of RNA-binding proteins (RBPs) but RNA-binding preferences for relatively few RBPs have been well defined. Current techniques for determining RNA targets, including in vitro selection and RNA co-immunoprecipitation, require significant time and labor investment. Here we introduce RNAcompete, a method for the systematic analysis of RNA binding specificities that uses a single binding reaction to determine the relative preferences of RBPs for short RNAs that contain a complete range of k-mers in structured and unstructured RNA contexts. We tested RNAcompete by analyzing nine diverse RBPs (HuR, Vts1, FUSIP1, PTB, U1A, SF2/ASF, SLM2, RBM4 and YB1). RNAcompete identified expected and previously unknown RNA binding preferences. Using in vitro and in vivo binding data, we demonstrate that preferences for individual 7-mers identified by RNAcompete are a more accurate representation of binding activity than are conventional motif models. We anticipate that RNAcompete will be a valuable tool for the study of RNA-protein interactions.
MeSH terms
Animals; Base Sequence; Binding Sites; Databases, Nucleic Acid; Genome; Molecular Sequence Data; Oligonucleotide Array Sequence Analysis; RNA; RNA-Binding Proteins; ROC Curve; Substrate Specificity
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