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Cross-linking, ligation, and sequencing of hybrids reveals RNA–RNA interactions in yeast
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Many protein-protein and protein-nucleic acid interactions have been experimentally characterized, whereas RNA-RNA interactions have generally only been predicted computationally. Here, we describe a high-throughput method to identify intramolecular and intermolecular RNA-RNA interactions experimentally by cross-linking, ligation, and sequencing of hybrids (CLASH). As validation, we identified 39 known target sites for box C/D modification-guide small nucleolar RNAs (snoRNAs) on the yeast pre-rRNA. Novel snoRNA-rRNA hybrids were recovered between snR4-5S and U14-25S. These are supported by native electrophoresis and consistent with previously unexplained data. The U3 snoRNA was found to be associated with sequences close to the 3' side of the central pseudoknot in 18S rRNA, supporting a role in formation of this structure. Applying CLASH to the yeast U2 spliceosomal snRNA led to a revised predicted secondary structure, featuring alternative folding of the 3' domain and long-range contacts between the 3' and 5' domains. CLASH should allow transcriptome-wide analyses of RNA-RNA interactions in many organisms.
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“early CLASH publications showed the utility of the chimeric analysis approach in mapping snoRNA interactions in yeast (Kudla et al., 2011) and more recently was utilized to generate an extensive catalog of putative snoRNA interactions and reveal novel targets of orphan snoRNAs (Gumienny et al., 2017).”
From Scalable and deep profiling of mRNA targets for individual microRNAs with chimeric eCLIP (2022) · cites this paper as Kudla et al., 2011
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Metadata
- Authors
- Kudla, G., Granneman, S., Hahn, D., Beggs, J. D., & Tollervey, D.
- Affiliations
- Wellcome Centre for Cell Biology (GB)
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2011-05-24
- Volume
- 108
- Issue
- 24
- Pages
- 10010-10015
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC3116431
- OpenAlex
- W2117165052
- Cited by
- 308
- References
- 39
- Citations by year
- 2026: 12 · 2025: 16 · 2024: 17 · 2023: 23 · 2022: 22 · 2021: 22 · 2020: 25 · 2019: 16 · 2018: 37 · 2017: 25 · 2016: 32 · 2015: 22 · 2014: 12 · 2013: 16 · 2012: 9
- FWCI
- 5.07
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Edinburgh Research Explorer (University of Edinburgh) (submittedVersion, other-oa)
- Funders
- Wellcome Trust
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
- Keywords
- RNA-RNA interactions, CLASH, box C/D snoRNA, small nucleolar RNA, pre-rRNA, U3 snoRNA, U2 snRNA, spliceosome, RNA secondary structure, Saccharomyces cerevisiae
- MeSH terms
- Base Sequence, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, RNA, Fungal, RNA, Ribosomal, RNA, Ribosomal, 18S, RNA, Small Nuclear, Saccharomyces cerevisiae, Blotting, Northern, Spliceosomes, Sequence Analysis, DNA, DNA, Complementary, RNA, Small Nucleolar
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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