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LC/MS analysis of cellular RNA reveals NAD-linked RNA

Nature Chemical Biology volume 5, pages 879–881 (2009)Cite this article

Abstract

We developed a general method to detect cellular small molecule–RNA conjugates that does not rely on the reactivity of the small molecule. This technique revealed NAD-linked RNA in Escherichia coli and Streptomyces venezuelae. Subsequent characterization showed that NAD is a 5′ modification of RNA, cannot be installed in vitro through aberrant transcriptional initiation, is only found among smaller cellular RNAs and is present at a surprisingly high abundance of ∼3,000 copies per cell.

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Figure 1: Discovery of a small molecule–linked nucleotide of [M-H]− m/z = 540.0533 from E. coli and S. venezuelae RNA.
Figure 2: Characterization of NAD-RNA.

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Acknowledgements

This work was supported by the Howard Hughes Medical Institute and the US National Institute of General Medical Sciences, National Institutes of Health (R01GM065865). Y.G.C. and W.E.K. gratefully acknowledge US National Science Foundation Graduate Research Fellowships.

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Author notes

  1. Y Grace Chen and Walter E Kowtoniuk: These authors contributed equally to this work.

Authors and Affiliations

  1. Howard Hughes Medical Institute and the Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA

    Y Grace Chen, Walter E Kowtoniuk, Isha Agarwal, Yinghua Shen & David R Liu

Authors
  1. Y Grace Chen
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  2. Walter E Kowtoniuk
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  3. Isha Agarwal
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  4. Yinghua Shen
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  5. David R Liu
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Contributions

Y.G.C., W.E.K. and D.R.L. designed research; Y.G.C., W.E.K., I.A. and Y.S. performed research; Y.G.C., W.E.K. and I.A. contributed new reagents and analytic tools; Y.G.C., W.E.K., I.A., Y.S. and D.R.L. analyzed data; and Y.G.C., W.E.K., Y.S. and D.R.L. wrote the paper.

Corresponding author

Correspondence to David R Liu.

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Chen, Y., Kowtoniuk, W., Agarwal, I. et al. LC/MS analysis of cellular RNA reveals NAD-linked RNA. Nat Chem Biol 5, 879–881 (2009). https://doi.org/10.1038/nchembio.235

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