Item Type: | Article |
---|---|
Title: | The mRNA-bound proteome and its global occupancy profile on protein-coding transcripts |
Creators Name: | Baltz, A.G. and Munschauer, M. and Schwanhaeusser, B. and Vasile, A. and Murakawa, Y. and Schueler, M. and Youngs, N. and Penfold-Brown, D. and Drew, K. and Milek, M. and Wyler, E. and Bonneau, R. and Selbach, M. and Dieterich, C. and Landthaler, M. |
Abstract: | Protein-RNA interactions are fundamental to core biological processes, such as mRNA splicing, localization, degradation, and translation. We developed a photoreactive nucleotide-enhanced UV crosslinking and oligo(dT) purification approach to identify the mRNA-bound proteome using quantitative proteomics and to display the protein occupancy on mRNA transcripts by next-generation sequencing. Application to a human embryonic kidney cell line identified close to 800 proteins. To our knowledge, nearly one-third were not previously annotated as RNA binding, and about 15% were not predictable by computational methods to interact with RNA. Protein occupancy profiling provides a transcriptome-wide catalog of potential cis-regulatory regions on mammalian mRNAs and showed that large stretches in 3' UTRs can be contacted by the mRNA-bound proteome, with numerous putative binding sites in regions harboring disease-associated nucleotide polymorphisms. Our observations indicate the presence of a large number of mRNA binders with diverse molecular functions participating in combinatorial posttranscriptional gene-expression networks. |
Keywords: | Binding Sites, Cell Line, Mass Spectrometry, Messenger RNA, Proteomics, RNA-Binding Proteins, RNA Sequence Analysis |
Source: | Molecular Cell |
ISSN: | 1097-2765 |
Publisher: | Cell Press |
Volume: | 46 |
Number: | 5 |
Page Range: | 674-690 |
Date: | 8 June 2012 |
Official Publication: | https://doi.org/10.1016/j.molcel.2012.05.021 |
PubMed: | View item in PubMed |
Repository Staff Only: item control page