Mapping exosome-substrate interactions in vivo by UV cross-linking

Clémentine Delan-Forino, David Tollervey

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract / Description of output

The RNA exosome complex functions in both the accurate processing and rapid degradation of many classes of RNA in eukaryotes and Archaea. Functional and structural analyses indicate that RNA can either be threaded through the central channel of the exosome or more directly access the active sites of the ribonucleases Rrp44 and Rrp6, but in most cases, it remains unclear how many substrates follow each pathway in vivo. Here we describe the method for using an UV cross-linking technique termed CRAC to generate stringent, transcriptome-wide mapping of exosome-substrate interaction sites in vivo and at base-pair resolution.

Original languageEnglish
Title of host publicationThe Eukaryotic RNA Exosome
Subtitle of host publicationMethods and Protocols
PublisherSpringer Nature
Pages105-126
Number of pages22
Volume2062
ISBN (Electronic)978-1-4939-9822-7
ISBN (Print)978-1-4939-9821-0
DOIs
Publication statusPublished - 2020

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherHumana Press
ISSN (Print)1064-3745

Keywords / Materials (for Non-textual outputs)

  • exosome
  • RNA processing
  • RNA degradation
  • Protein-RNA interaction
  • RNA-binding sites
  • UV cross-linking
  • yeast

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