Transcriptomics

Dataset Information

238

The PIN domain endonuclease Utp24 cleaves pre-ribosomal RNA at two coupled sites in yeast and humans


ABSTRACT: Purpose: Pre-ribosomal RNA is cleaved at defined sites, but many endonucleases involved in 18S rRNA release are not known. We apply an in vivo cross-linking technique coupled with deep sequencing (CRAC) that captures transcriptome-wide interactions between a yeast candidate pre-rRNA endonuclease (Utp24) and its targets in a living cell. Methods: We apply CRAC to an HTP-tagged Utp24 protein (HTP: His6 - TEV cleavage site - two copies of the z-domain of Protein A). At least two independent experiments were performed and analyzed separately. Results: We found that yeast Utp24 UV-crosslinked in vivo to the U3 snoRNA and all (pre-)rRNA elements that form the central pseudoknot in the 18S rRNA. The pseudoknot is an evolutionarily highly conserved structure that is required to ensure pre-rRNA processing at three cleavage sites (A0, A1 and A2) and still present in the mature rRNA. According to our crosslinking data, the endonuclease Utp24 is placed in close proximity to site A1 at the 5'-end of the 18S rRNA. Conclusion: Our study strongly supports the hypothesis that Utp24 cleaves pre-rRNA at sites A1 and A2. Examination of targets for pre-rRNA endonucleases in yeast cells.

ORGANISM(S): Saccharomyces cerevisiae  

SUBMITTER: Franziska Weichmann   David Colvin  Claudia Schneider  David Tollervey  Katherine E Sloan  Nicholas J Watkins  Graeme R Wells  Grzegorz Kudla 

PROVIDER: E-GEOD-75991 | ArrayExpress | 2016-04-01

SECONDARY ACCESSION(S): GSE75991SRP067353PRJNA305911

REPOSITORIES: GEO, ArrayExpress, ENA

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