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. 2012 Jul 17;13(7):163.
doi: 10.1186/gb4030.

The greatest catch: big game fishing for mRNA-bound proteins

Christopher R Sibley, Jan Attig, Jernej Ule

The greatest catch: big game fishing for mRNA-bound proteins

Christopher R Sibley et al. Genome Biol. .

Abstract

Purification of proteins cross-linked to mRNAs has identified 800 mRNA-binding proteins and their characteristics.

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Figures

Figure 1
Figure 1
dentification of the mRNA interactome. RNA-binding proteins are covalently cross-linked to RNAs using 254 nM UV-C light [2] or 365 nM UV light in conjunction with photoactivatable ribonucleosides (PARs) such as 4-thiouridine (4SU) [2,3]. The RNA is used as bait in a pull-down with oligo-dT-coated beads and purified under stringent conditions to eliminate contamination from non-cross-linked proteins. (a) Proteins are released by RNAse digestion and analyzed by mass spectrometry. (b) RNAs are blotted onto nitrocellulose, released by proteinase K, and analyzed with high-throughput sequencing. The diagnostic T to C changes in 4SU-labelled RNA identifies cross-link sites. PAR-CLIP, photoactivatable ribonucleoside UV cross-linked immunoprecipitation; X-link, cross-link.
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References

    1. Cooper TA, Wan L, Dreyfuss G. RNA and disease. Cell. 2009;136:777–793. doi: 10.1016/j.cell.2009.02.011. - DOI - PMC - PubMed
    1. Castello A, Fischer B, Eichelbaum K, Horos R, Beckmann BM, Strein C, Davey NE, Humphreys DT, Preiss T, Steinmetz LM, Krijgsveld J, Hentze MW. Insights into RNA biology from an atlas of mammalian mRNA-binding proteins. Cell. 2012;149:1393–1406. doi: 10.1016/j.cell.2012.04.031. - DOI - PubMed
    1. Baltz AG, Munschauer M, Schwanhausser B, Vasile A, Murakawa Y, Schueler M, Youngs N, Penfold-Brown D, Drew K, Milek M, Wyler E, Bonneau R, Selbach M, Dieterich C, Landthaler M. The mRNA-bound proteome and its global occupancy profile on protein-coding transcripts. Mol Cell. 2012;46:674–690. doi: 10.1016/j.molcel.2012.05.021. - DOI - PubMed
    1. Ule J, Jensen KB, Ruggiu M, Mele A, Ule A, Darnell RB. CLIP identifies Nova-regulated RNA networks in the brain. Science. 2003;302:1212–1215. doi: 10.1126/science.1090095. - DOI - PubMed
    1. Hafner M, Landthaler M, Burger L, Khorshid M, Hausser J, Berninger P, Rothballer A, Ascano M, Jungkamp AC, Munschauer M, Ulrich A, Wardle GS, Dewell S, Zavolan M, Tuschl T. Transcriptome-wide identification of RNA-binding protein and microRNA target sites by PAR-CLIP. Cell. 2010;141:129–141. doi: 10.1016/j.cell.2010.03.009. - DOI - PMC - PubMed

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