Poster abstracts
Poster number 28 submitted by Guramrit Singh
Chemical crosslinking enhances RNA immunoprecipitation for efficient identification of binding sites of proteins that photo-crosslink poorly with RNA
Robert Patton (Department of Physics, Center for RNA Biology, The Ohio State University, Columbus, OH 43210), Lauren A. Woodward, Justin W. Mabin (Department of Molecular Genetics, Center for RNA Biology, The Ohio State University, Columbus, OH 43210), Ralf Bundschuh (Department of Physics, Division of Hematology, Department of Internal Medicine, Center for RNA Biology, The Ohio State University, Columbus, OH 43210), Guramrit Singh (Department of Molecular Genetics, Center for RNA Biology, The Ohio State University, Columbus, OH 43210)
Abstract:
In all eukaryotic cells, RNA binding proteins (RBPs) regulate RNA activity to control cellular function. To fulfill this function, RBPs often directly contact RNA. The chemical contacts between RNA and many RBPs can be covalently crosslinked upon exposure to ultraviolet (UV) light. This photo-crosslinking property has been exploited to reveal the complement of RBPs that directly bind RNA in eukaryotic cells. The same characteristic underlies the widely used UV cross-linking and immunoprecipitation followed by sequencing (CLIP-Seq) approach to identify binding sites of an RBP. Here we show that the human genome encodes several hundred proteins that act on RNA but UV crosslink poorly with it because they may not be in direct physical contact with RNA. These
Keywords: RNA binding proteins, exon junction complex, RIPiT-Seq