Abstract
Background. Uropathogenic Escherichia coli (UPEC), a leading cause of urinary tract and invasive infections worldwide, is rapidly acquiring multidrug resistance, hastening the need for selective new anti-infective agents. Here we demonstrate the molecular target of DU011, our previously discovered potent, nontoxic, small-molecule inhibitor of UPEC polysaccharide capsule biogenesis and virulence. Methods. Real-time polymerase chain reaction analysis and a target-overexpression drug-suppressor screen were used to localize the putative inhibitor target. A thermal shift assay quantified interactions between the target protein and the inhibitor, and a novel DNase protection assay measured chemical inhibition of protein-DNA interactions. Virulence of a regulatory target mutant was assessed in a murine sepsis model. Results. MprA, a MarR family transcriptional repressor, was identified as the putative target of the DU011 inhibitor. Thermal shift measurements indicated the formation of a stable DU011-MprA complex, and DU011 abrogated MprA binding to its DNA promoter site. Knockout of mprA had effects similar to that of DU011 treatment of wild-type bacteria: a loss of encapsulation and complete attenuation in a murine sepsis model, without any negative change in antibiotic resistance. Conclusions. MprA regulates UPEC polysaccharide encapsulation, is essential for UPEC virulence, and can be targeted without inducing antibiotic resistance.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1330-1339 |
| Number of pages | 10 |
| Journal | Journal of Infectious Diseases |
| Volume | 213 |
| Issue number | 8 |
| DOIs | |
| State | Published - Apr 15 2016 |
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Keywords
- E. Coli
- multidrug efflux pumps
- polysaccharide capsule
- small-molecule capsule inhibitor
ASJC Scopus subject areas
- Immunology and Allergy
- Infectious Diseases
Cite this
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Threading the Needle : Small-Molecule Targeting of a Xenobiotic Receptor to Ablate Escherichia coli Polysaccharide Capsule Expression Without Altering Antibiotic Resistance. / Arshad, Mehreen; Goller, Carlos C.; Pilla, Danielle; Schoenen, Frank J.; Seed, Patrick C.
In: Journal of Infectious Diseases, Vol. 213, No. 8, 15.04.2016, p. 1330-1339.Research output: Contribution to journal › Article
TY - JOUR
T1 - Threading the Needle
T2 - Small-Molecule Targeting of a Xenobiotic Receptor to Ablate Escherichia coli Polysaccharide Capsule Expression Without Altering Antibiotic Resistance
AU - Arshad, Mehreen
AU - Goller, Carlos C.
AU - Pilla, Danielle
AU - Schoenen, Frank J.
AU - Seed, Patrick C.
PY - 2016/4/15
Y1 - 2016/4/15
N2 - Background. Uropathogenic Escherichia coli (UPEC), a leading cause of urinary tract and invasive infections worldwide, is rapidly acquiring multidrug resistance, hastening the need for selective new anti-infective agents. Here we demonstrate the molecular target of DU011, our previously discovered potent, nontoxic, small-molecule inhibitor of UPEC polysaccharide capsule biogenesis and virulence. Methods. Real-time polymerase chain reaction analysis and a target-overexpression drug-suppressor screen were used to localize the putative inhibitor target. A thermal shift assay quantified interactions between the target protein and the inhibitor, and a novel DNase protection assay measured chemical inhibition of protein-DNA interactions. Virulence of a regulatory target mutant was assessed in a murine sepsis model. Results. MprA, a MarR family transcriptional repressor, was identified as the putative target of the DU011 inhibitor. Thermal shift measurements indicated the formation of a stable DU011-MprA complex, and DU011 abrogated MprA binding to its DNA promoter site. Knockout of mprA had effects similar to that of DU011 treatment of wild-type bacteria: a loss of encapsulation and complete attenuation in a murine sepsis model, without any negative change in antibiotic resistance. Conclusions. MprA regulates UPEC polysaccharide encapsulation, is essential for UPEC virulence, and can be targeted without inducing antibiotic resistance.
AB - Background. Uropathogenic Escherichia coli (UPEC), a leading cause of urinary tract and invasive infections worldwide, is rapidly acquiring multidrug resistance, hastening the need for selective new anti-infective agents. Here we demonstrate the molecular target of DU011, our previously discovered potent, nontoxic, small-molecule inhibitor of UPEC polysaccharide capsule biogenesis and virulence. Methods. Real-time polymerase chain reaction analysis and a target-overexpression drug-suppressor screen were used to localize the putative inhibitor target. A thermal shift assay quantified interactions between the target protein and the inhibitor, and a novel DNase protection assay measured chemical inhibition of protein-DNA interactions. Virulence of a regulatory target mutant was assessed in a murine sepsis model. Results. MprA, a MarR family transcriptional repressor, was identified as the putative target of the DU011 inhibitor. Thermal shift measurements indicated the formation of a stable DU011-MprA complex, and DU011 abrogated MprA binding to its DNA promoter site. Knockout of mprA had effects similar to that of DU011 treatment of wild-type bacteria: a loss of encapsulation and complete attenuation in a murine sepsis model, without any negative change in antibiotic resistance. Conclusions. MprA regulates UPEC polysaccharide encapsulation, is essential for UPEC virulence, and can be targeted without inducing antibiotic resistance.
KW - E. Coli
KW - multidrug efflux pumps
KW - polysaccharide capsule
KW - small-molecule capsule inhibitor
UR - http://www.scopus.com/inward/record.url?scp=84965128664&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84965128664&partnerID=8YFLogxK
U2 - 10.1093/infdis/jiv584
DO - 10.1093/infdis/jiv584
M3 - Article
C2 - 26671885
AN - SCOPUS:84965128664
VL - 213
SP - 1330
EP - 1339
JO - Journal of Infectious Diseases
JF - Journal of Infectious Diseases
SN - 0022-1899
IS - 8
ER -