Visualization of the Nucleolus Using 5' Ethynyl Uridine
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- Keywords
- Click-iT, Confocal Microscopy, Ethynyl Uridine, Nucleolus, Transcription,
- MeSH
- Cell Nucleolus * metabolism MeSH
- Microscopy MeSH
- Ribosomes metabolism MeSH
- RNA * metabolism MeSH
- Uridine metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- RNA * MeSH
- Uridine MeSH
Labeling of the nucleolus in Arabidopsis thaliana can be achieved by incorporation of 5'-ethynyl uridine (EU) into bulk RNA. Although EU does not selectively label the nucleolus, the abundance of ribosomal transcripts results in the predominant accumulation of the signal in the nucleolus. Ethynyl uridine has the advantage of being detected via Click-iT chemistry providing a specific signal and low background. While the protocol presented here employs fluorescent dye and allows visualization of the nucleolus by microscopy, this method can also be used for other downstream applications. Though we tested nucleolar labeling only in A. thaliana, in principle it can be applied to other plant species.
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Li Q, Kim Y, Namm J et al (2006) RNA-selective, live cell imaging probes for studying nuclear structure and function. Chem Biol 13:615–623. https://doi.org/10.1016/j.chembiol.2006.04.007 PubMed DOI
Pendle AF, Clark GP, Boon R et al (2005) Proteomic analysis of the Arabidopsis nucleolus suggests novel nucleolar functions. Mol Biol Cell 16:260–269. https://doi.org/10.1091/mbc.e04-09-0791 PubMed DOI PMC
Dvořáčková M, Fajkus J (2018) Visualization of the nucleolus using ethynyl uridine. Front Plant Sci 9:177. https://doi.org/10.3389/fpls.2018.00177 PubMed DOI PMC
Breinbauer R, Köhn M (2003) Azide-alkyne coupling – a powerful reaction for bioconjugate chemistry. Chembiolchem 4:1147–1149 DOI
Uttamapinant C, Tangpeerachaikul A, Grecian S et al (2012) Fast, cell-compatible click chemistry with copper-chelating azides for biomolecular labeling. Angew Chem Int Ed Engl 51:5852–5856. https://doi.org/10.1002/anie.201108181 PubMed DOI PMC
Kotogány E, Dudits D, Horváth GV, Ayaydin F (2010) A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine. Plant Methods 6:5. https://doi.org/10.1186/1746-4811-6-5 PubMed DOI PMC