Genomic Deoxyxylulose
Phosphate Reductoisomerase (DXR)
Mutations Conferring Resistance to the Antimalarial Drug Fosmidomycin
in <i>E. coli</i>
Gur Pines
Eun Joong Oh
Marcelo C. Bassalo
Alaksh Choudhury
Andrew D. Garst
Reilly G. Fankhauser
Carrie A. Eckert
Ryan T. Gill
10.1021/acssynbio.8b00219.s001
https://acs.figshare.com/articles/journal_contribution/Genomic_Deoxyxylulose_Phosphate_Reductoisomerase_DXR_Mutations_Conferring_Resistance_to_the_Antimalarial_Drug_Fosmidomycin_in_i_E_coli_i_/7436303
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 <i>ispC</i> gene in its native genomic context in <i>Escherichia
coli</i>. 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 <i>ispC</i> gene product, DXR. This selection is especially challenging since
it is relatively weak in <i>E. coli</i>, 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 <i>Plasmodium falciparum</i>. This approach
may have implications for the isolation of resistance-conferring mutations
and may affect the design of future generations of fosmidomycin-based
drugs.
2018-11-21 00:00:00
Antimalarial Drug Fosmidomycin
E . coli Sequence
E . coli
site
technology
ispC gene product
CREATE
Mutations Conferring Resistance
generation
Genomic Deoxyxylulose Phosphate Reductoisomerase
mutation
resistance
ligand binding pocket
activity mapping technologies
trackable genome engineering
CRISPR
. coli
antimalarial drug fosmidomycin
isolation
DXR
malaria-inducing Plasmodium falciparum