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Chemical genetic interrogation of natural variation uncovers a molecule that is glycoactivated

Nature Chemical Biology volume 3, pages 716–721 (2007)Cite this article

Abstract

Natural variation in human drug metabolism and target genes can cause pharmacogenetic or interindividual variation in drug sensitivity1,2. We reasoned that natural pharmacogenetic variation in model organisms could be systematically exploited to facilitate the characterization of new small molecules. To test this, we subjected multiple Arabidopsis thaliana accessions to chemical genetic screens and discovered 12 accession-selective hit molecules. As a model for understanding this variation, we characterized natural resistance to hypostatin, a new inhibitor of cell expansion. Map-based cloning identified HYR1, a UDP glycosyltransferase (UGT), as causative for hypostatin resistance. Multiple lines of evidence demonstrate that HYR1 glucosylates hypostatin in vivo to form a bioactive glucoside. Additionally, we delineated a HYR1 substrate motif and used it to identify another molecule modulated by glucosylation. Our results demonstrate that natural variation can be exploited to inform the biology of new small molecules, and that UGT sequence variation affects xenobiotic sensitivity across biological kingdoms.

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Figure 1: Natural variation in small-molecule sensitivity exists between A. thaliana accessions.
Figure 2: The natural hypostatin resistance locus HYR1 is a UGT1A1 homolog.
Figure 3: Glucosylation activates hypostatin.
Figure 4: HYR1 modulates sensitivity to multiple xenobiotics.
Figure 5: An SNP explains most natural variation in hypostatin sensitivity.

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Acknowledgements

This work was supported by a Natural Sciences and Engineering Research Council (NSERC) of Canada Discovery grant, a Canadian Research Chair and a Canadian Foundation for Innovation startup grant to S.R.C., and an NSERC postgraduate fellowship to R.S.P. We thank P. McCourt and P. Roy for discussion and comments on the manuscript; A. Young of the University of Toronto Mass Spectrometry Facility for MS analyses; J. Coleman, R. Sage and the University of Toronto Department of Botany for purchasing compounds; and T. Kwok and P. Roy for tests on C. elegans. Lastly, we thank an anonymous reviewer for alerting us to ref. 20.

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Authors and Affiliations

  1. Department of Cell and Systems Biology, University of Toronto, 25 Willcocks St., Toronto, M5S 3B2, Ontario, Canada

    Yang Zhao, Tszfung F Chow, Rachel S Puckrin & Simon E Alfred

  2. Department of Chemistry, University of California-Riverside, 501 Big Springs Road, Riverside, 92521, California, USA

    Albert K Korir & Cynthia K Larive

  3. Center for Plant Cell Biology and the Department of Botany and Plant Sciences, University of California-Riverside, 2150 Batchelor Hall, Riverside, 92521, California, USA

    Sean R Cutler

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  1. Yang Zhao
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  2. Tszfung F Chow
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Contributions

Y.Z. performed the Chembridge natural variation screen, cloned HYR1 and characterized hypostatin. T.F.C. performed the natural variation screens of 82 novel bioactive compounds, with assistance from R.S.P. and S.E.A. for the molecules chuboxypyr and polarazine. A.K.K. performed and interpreted, and C.K.L. supervised, the hypostatin LC-MS studies. S.R.C. conceived the project, assisted in experimental design and wrote the manuscript.

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Correspondence to Sean R Cutler.

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Supplementary Figures 1–2, Supplementary Tables 1–6 and Supplementary Methods (PDF 328 kb)

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Zhao, Y., Chow, T., Puckrin, R. et al. Chemical genetic interrogation of natural variation uncovers a molecule that is glycoactivated. Nat Chem Biol 3, 716–721 (2007). https://doi.org/10.1038/nchembio.2007.32

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