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| Status |
Public on Jan 11, 2017 |
| Title |
In vivo RNA binding sites of the yeast ribosome biogenesis factor Utp23 |
| Organism |
Saccharomyces cerevisiae |
| Experiment type |
Other
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| Summary |
Purpose: Pre-ribosomal RNA is cleaved at defined sites to release the mature ribosomal RNAs, but the functions of many ribosome biogenesis factors 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 the yeast PIN domain protein Utp23 and its targets in a living cell.
Methods: We apply CRAC to an HTP-tagged Utp23 protein (HTP: His6 - TEV cleavage site - two copies of the z-domain of Protein A) in budding yeast. At least two independent experiments were performed and analysed separately. A non-tagged yeast strain was also used as a negative control.
Results: We found that yeast Utp23 UV-crosslinked in vivo to the snR30 snoRNA and to the eukaryotic-specific expansion segment 6 (ES6) in the 18S rRNA.
Conclusion: According to our crosslinking data, Utp23 is perfectly positioned to coordinate release of the snR30 snoRNA from the 18S ES6 region.
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| Overall design |
Examination of targets for RNA-binding ribosome biogenesis factors in yeast cells.
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| Contributor(s) |
Wells GR, Weichmann F, Sloan KE, Colvin D, Watkins NJ, Schneider C |
| Citation(s) |
28082392 |
| Submission date |
Sep 22, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Claudia Schneider |
| E-mail(s) |
Claudia.Schneider@ncl.ac.uk
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| Organization name |
Newcastle University
|
| Street address |
Framlington Place
|
| City |
Newcastle upon Tyne |
| ZIP/Postal code |
NE2 4HH |
| Country |
United Kingdom |
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| Platforms (2) |
| GPL13272 |
Illumina Genome Analyzer IIx (Saccharomyces cerevisiae) |
| GPL13821 |
Illumina HiSeq 2000 (Saccharomyces cerevisiae) |
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| Samples (3) |
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| Relations |
| BioProject |
PRJNA343936 |
| SRA |
SRP090308 |
