ArticleAnalytical and bioanalytical chemistry2017
Novel ribonuclease activity of cusativin from Cucumis sativus for mapping nucleoside modifications in RNA.
Article in Analytical and bioanalytical chemistry, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 40 citations in OpenAlex.
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- Human 28S rRNA analysed by state-of-the-art oligonucleotide mass spectrometry: benchmarking current capabilities and a call to action for MS-Seq.bioRxiv : the preprint server for biology · 2026Article
- tRNA modifications: greasing the wheels of translation and beyond.RNA biology · 2025Review
- Analysis of RNA and Its Modifications.Annual review of analytical chemistry (Palo Alto, Calif.) · 2024Review
- Tyrosine transfer RNA levels and modifications during blood-feeding and vitellogenesis in the mosquito,bioRxiv : the preprint server for biology · 2023Article
- Direct sequencing of totalRNA (New York, N.Y.) · 2023Article
- Challenges and emerging trends in liquid chromatography-based analyses of mRNA pharmaceuticals.Journal of pharmaceutical and biomedical analysis · 2023Review
- Human RNase 4 improves mRNA sequence characterization by LC-MS/MS.Nucleic acids research · 2022Article
- RNA Cleavage Properties of Nucleobase-Specific RNase MC1 and Cusativin Are Determined by the Dinucleotide-Binding Interactions in the Enzyme-Active Site.International journal of molecular sciences · 2022Article
- Identification and mapping of post-transcriptional modifications on the HIV-1 antisense transcriptRNA (New York, N.Y.) · 2022Article
- The nature of the modification at position 37 of tRNAPhe correlates with acquired taxol resistance.Nucleic acids research · 2021Article
- Locating chemical modifications in RNA sequences through ribonucleases and LC-MS based analysis.Methods in enzymology · 2021Article
- Characterization of UVA-Induced Alterations to Transfer RNA Sequences.Biomolecules · 2020Article
- Naturally occurring modified ribonucleosides.Wiley interdisciplinary reviews. RNA · 2020Review
- 2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications.Journal of visualized experiments : JoVE · 2020Article
- Improved RNA modification mapping of cellular non-coding RNAs using C- and U-specific RNases.The Analyst · 2020Article
- A general LC-MS-based RNA sequencing method for direct analysis of multiple-base modifications in RNA mixtures.Nucleic acids research · 2019Article
- tRNA Modification Profiles and Codon-Decoding Strategies in Methanocaldococcus jannaschii.Journal of bacteriology · 2019Article
- Improved application of RNAModMapper - An RNA modification mapping software tool - For analysis of liquid chromatography tandem mass spectrometry (LC-MS/MS) data.Methods (San Diego, Calif.) · 2019Article
- Detection of ribonucleoside modifications by liquid chromatography coupled with mass spectrometry.Biochimica et biophysica acta. Gene regulatory mechanisms · 2019Review
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
A recombinant ribonuclease, cusativin, was characterized for its cytidine-specific cleavage ability of RNA to map chemical modifications. Following purification of native cusativin protein as described before (Rojo et al. Planta 194:328, 17), partial amino acid sequencing was carried out to identify the corresponding protein coding gene in cucumber genome. Cloning and heterologous expression of the identified gene in Escherichia coli resulted in successful production of active protein as a C-terminal His-tag fusion protein. The ribonuclease activity and cleavage specificity of the fusion protein were confirmed with a variety of tRNA isoacceptors and total tRNA. Characterization of cusativin digestion products by ion-pairing reverse-phase liquid chromatography coupled with mass spectrometry (IP-RP-LC-MS) analysis revealed cleavage of CpA, CpG, and CpU phosphodiester bonds at the 3'-terminus of cytidine under optimal digestion conditions. Ribose methylation or acetylation of cytosine inhibited RNA cleavage. The CpC phosphodiester bond was also resistant to cusativin-mediated RNA cleavage; a feature to our knowledge has not been reported for other nucleobase-specific ribonucleases. Here, we demonstrate the analytical utility of such a novel feature for obtaining high-sequence coverage and accurate mapping of modified residues in substrate RNAs. Graphical abstract Cytidine-specific novel ribonuclease activity of cusativin.
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