Urdaneta, Erika C. and Vieira-Vieira, Carlos H. and Hick, Timon and Wessels, Hans-Herrmann and Figini, Davide and Moschall, Rebecca and Medenbach, Jan and Ohler, Uwe and Granneman, Sander and Selbach, Matthias and Beckmann, Benedikt M. (2019) Purification of cross-linked RNA-protein complexes by phenol-toluol extraction. NATURE COMMUNICATIONS, 10: 990. ISSN 2041-1723,
Full text not available from this repository. (Request a copy)Abstract
Recent methodological advances allowed the identification of an increasing number of RNA-binding proteins (RBPs) and their RNA-binding sites. Most of those methods rely, however, on capturing proteins associated to polyadenylated RNAs which neglects RBPs bound to non-adenylate RNA classes (tRNA, rRNA, pre-mRNA) as well as the vast majority of species that lack poly-A tails in their mRNAs (including all archea and bacteria). We have developed the Phenol Toluol extraction (PTex) protocol that does not rely on a specific RNA sequence or motif for isolation of cross-linked ribonucleoproteins (RNPs), but rather purifies them based entirely on their physicochemical properties. PTex captures RBPs that bind to RNA as short as 30 nt, RNPs directly from animal tissue and can be used to simplify complex workflows such as PAR-CLIP. Finally, we provide a global RNA-bound proteome of human HEK293 cells and the bacterium Salmonella Typhimurium.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | BINDING PROTEIN; MESSENGER-RNA; FUNCTIONAL-ANALYSIS; EXOSOME; LINKING; CLIP; IDENTIFICATION; REVEALS; SITES; DNA; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 16 Apr 2020 11:10 |
| Last Modified: | 16 Apr 2020 11:10 |
| URI: | https://pred.uni-regensburg.de/id/eprint/27479 |
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