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Transcriptome-wide discovery of coding and noncoding RNA-binding proteins.
Huang, Rongbing; Han, Mengting; Meng, Liying; Chen, Xing.
Afiliación
  • Huang R; College of Chemistry and Molecular Engineering, Peking University, 100871 Beijing, China.
  • Han M; Beijing National Laboratory for Molecular Sciences, 100871 Beijing, China.
  • Meng L; College of Chemistry and Molecular Engineering, Peking University, 100871 Beijing, China.
  • Chen X; Beijing National Laboratory for Molecular Sciences, 100871 Beijing, China.
Proc Natl Acad Sci U S A ; 115(17): E3879-E3887, 2018 04 24.
Article en En | MEDLINE | ID: mdl-29636419
Transcriptome-wide identification of RNA-binding proteins (RBPs) is a prerequisite for understanding the posttranscriptional gene regulation networks. However, proteomic profiling of RBPs has been mostly limited to polyadenylated mRNA-binding proteins, leaving RBPs on nonpoly(A) RNAs, including most noncoding RNAs (ncRNAs) and pre-mRNAs, largely undiscovered. Here we present a click chemistry-assisted RNA interactome capture (CARIC) strategy, which enables unbiased identification of RBPs, independent of the polyadenylation state of RNAs. CARIC combines metabolic labeling of RNAs with an alkynyl uridine analog and in vivo RNA-protein photocross-linking, followed by click reaction with azide-biotin, affinity enrichment, and proteomic analysis. Applying CARIC, we identified 597 RBPs in HeLa cells, including 130 previously unknown RBPs. These newly discovered RBPs can likely bind ncRNAs, thus uncovering potential involvement of ncRNAs in processes previously unknown to be ncRNA-related, such as proteasome function and intermediary metabolism. The CARIC strategy should be broadly applicable across various organisms to complete the census of RBPs.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Unión al ARN / Perfilación de la Expresión Génica / Transcriptoma Límite: Humans Idioma: En Revista: Proc Natl Acad Sci U S A Año: 2018 Tipo del documento: Article País de afiliación: China Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Unión al ARN / Perfilación de la Expresión Génica / Transcriptoma Límite: Humans Idioma: En Revista: Proc Natl Acad Sci U S A Año: 2018 Tipo del documento: Article País de afiliación: China Pais de publicación: Estados Unidos