Abstract
Dynemicin A is a member of a subfamily of enediyne antitumour antibiotics characterized by a 10-membered carbocycle fused to an anthraquinone, both of polyketide origin. Sequencing of the dynemicin biosynthetic gene cluster in Micromonospora chersina previously identified an enediyne polyketide synthase (PKS), but no anthraquinone PKS, suggesting gene(s) for biosynthesis of the latter were distant from the core dynemicin cluster. To identify these gene(s), we sequenced and analysed the genome of M. chersina. Sequencing produced a short list of putative PKS candidates, yet CRISPR-Cas9 mutants of each locus retained dynemicin production. Subsequently, deletion of two cytochromes P450 in the dynemicin cluster suggested that the dynemicin enediyne PKS, DynE8, may biosynthesize the anthraquinone. Together with 18O-labelling studies, we now present evidence that DynE8 produces the core scaffolds of both the enediyne and anthraquinone, and provide a working model to account for their formation from the programmed octaketide of the enediyne PKS.
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Acknowledgements
The authors acknowledge the Experimental and Computational Genomics Core at The Johns Hopkins University for sequencing and assembling the M. chersina genome, A. Majumdar, I.P. Mortimer, B. Steinberg, F.B. d'Andrea and C.L. Malbon for NMR, UPLC-ESI-MS, mechanical, synthesis and coding help, respectively, and A.L. Eller for performing preliminary experiments. The authors thank the following for donations: H. Ikeda for E. coli GM2929 hsdS::Tn10 (pUB307::Tn7), Y. Tong, T. Weber and S.Y. Lee for pCRISPR-Cas9 and R.F. Li for obtaining these items, and the Agricultural Research Service for M. chersina NRRL B-24756, J. Stuart at Omega Protein Corporation for fish meal and C. Fischer and J.C. Vederas for use of the O2 fermentation system. This work was supported by National Institutes of Health grants R01 ES001670 and T32 GM080189.
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D.R.C. and C.A.T. designed the experiments, analysed the data and wrote the manuscript. D.R.C. performed experiments.
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Cohen, D., Townsend, C. A dual role for a polyketide synthase in dynemicin enediyne and anthraquinone biosynthesis. Nature Chem 10, 231–236 (2018). https://doi.org/10.1038/nchem.2876
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DOI: https://doi.org/10.1038/nchem.2876
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