Identification of RNA-Binding Proteins Associated to RNA Structural Elements.
RNA motifs guide the interaction with specific proteins leading to the assembly of ribonucleoprotein complexes that perform key functions in cellular processes. Internal ribosome entry site (IRES) elements are organized in structural domains that determine internal initiation of translation. In this chapter we describe a pull-down assay using streptavidin-aptamer tagged RNAs that combines RNA structure-dependent protein isolation with proteomic analysis to identify novel interactors recognizing RNA structural domains. This approach takes advantage of tRNA-scaffold guided expression, allowing the identification of factors belonging to networks involved in RNA and protein metabolism.
['Fernandez-Chamorro|Javier|J|', 'Francisco-Velilla|Rosario|R|', 'Embarc-Buh|Azman|A|', 'Martinez-Salas|Encarnacion|E|']
D052157:Aptamers, Nucleotide D004591:Electrophoresis, Polyacrylamide Gel D006801:Humans D000067556:Internal Ribosome Entry Sites D013058:Mass Spectrometry D059372:Nucleotide Motifs D014176:Protein Biosynthesis D040901:Proteomics D012313:RNA D012343:RNA, Transfer D016601:RNA-Binding Proteins D019809:Streptavidin D013379:Substrate Specificity
RNA-binding proteins RNA structure Pull-down IRES elements Streptavidin aptamer Translation control tRNA scaffold