Multicopy suppressors for novel antibacterial compounds reveal targets and drug efflux susceptibility

被引:70
作者
Li, XM
Zolli-Juran, M
Cechetto, JD
Daigle, DM
Wright, GD
Brown, ED
机构
[1] McMaster Univ, Dept Biochem, Hamilton, ON L8N 3Z5, Canada
[2] McMaster Univ, Antimicrobial Res Ctr, Hamilton, ON L8N 3Z5, Canada
来源
CHEMISTRY & BIOLOGY | 2004年 / 11卷 / 10期
关键词
D O I
10.1016/j.chembiol.2004.08.014
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Gene dosage has frequently been exploited to select for genetic interactions between a particular mutant and clones from a random genomic library at high copy. We report here the first use of multicopy suppression as a forward genetic method to determine cellular targets and potential resistance mechanisms for novel antibacterial compounds identified through high-throughput screening. A screen of 8640 small molecules for growth inhibition of a hyperpermeable strain of Escherichia coli led to the identification of 49 leads for suppressor selection from clones harboring an E. coli genomic library. The majority of suppressors were found to encode the multidrug efflux pump AcrB, indicating that those compounds were substrates for efflux. Two leads, which produced clones containing the gene folA, encoding dihydrofolate reductase (DHFR), proved to target DHFR in vivo and were competitive inhibitors in vitro.
引用
收藏
页码:1423 / 1430
页数:8
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