Genomic Deoxyxylulose Phosphate Reductoisomerase (DXR) Mutations Conferring Resistance to the Antimalarial Drug Fosmidomycin in E. coli

Carrie Eckert, Gur Pines, Eun Oh, Marcelo Bassalo, Alaksh Choudhury, Andrew Garst, Reilly Fankhauser, Ryan Gill

Research output: Contribution to journalArticlepeer-review

10 Scopus Citations

Abstract

Sequence to activity mapping technologies are rapidly developing, enabling the generation and isolation of mutations conferring novel phenotypes. Here we used the CRISPR enabled trackable genome engineering (CREATE) technology to investigate the inhibition of the essential ispC gene in its native genomic context in Escherichia coli. We created a full saturation library of 33 sites proximal to the ligand binding pocket and challenged this library with the antimalarial drug fosmidomycin, which targets the ispC gene product, DXR. This selection is especially challenging since it is relatively weak in E. coli, with multiple naturally occurring pathways for resistance. We identified several previously unreported mutations that confer fosmidomycin resistance, in highly conserved sites that also exist in pathogens including the malaria-inducing Plasmodium falciparum. This approach may have implications for the isolation of resistance-conferring mutations and may affect the design of future generations of fosmidomycin-based drugs.
Original languageAmerican English
Pages (from-to)2824-2832
Number of pages9
JournalACS Synthetic Biology
Volume7
Issue number12
DOIs
StatePublished - 2018

NREL Publication Number

  • NREL/JA-2700-73080

Keywords

  • acquired resistance
  • CRISPR/Cas9
  • deoxyxylulose phosphate reductoisomerase
  • fosmidomycin
  • malaria
  • sequence to activity mapping

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