|
KMID : 0545120200300070982
|
|
Journal of Microbiology and Biotechnology
2020 Volume.30 No. 7 p.982 ~ p.995
|
|
|
Cloning and Functional Characterization of Putative Escherichia coli ABC Multidrug Efflux Transporter YddA
|
|
Feng Zhenyue
Liu Defu
Liu Ziwen
Liang Yimin
Wang Yanhong
Liu Qingpeng
Liu Zhenhua
Zang Zhongjing
Cui Yudong
|
|
|
Abstract
|
|
|
|
A putative multidrug efflux gene, yddA, was cloned from the Escherichia coli K-12 strain. A drugsensitive strain of E. coli missing the main multidrug efflux pump AcrB was constructed as a host and the yddA gene was knocked out in wild-type (WT) and drug-sensitive E. coliΔacrB to study the yddA function. Sensitivity to different substrates of WT E.coli, E. coliΔyddA, E. coliΔacrB and E. coliΔacrBΔyddA strains was compared with minimal inhibitory concentration (MIC) assays and fluorescence tests. MIC assay and fluorescence test results showed that YddA protein was a multidrug efflux pump that exported multiple substrates. Three inhibitors, ortho-vanadate, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and reserpine, were used in fluorescence tests. Ortho-vanadate and reserpine significantly inhibited the efflux and increased accumulation of ethidium bromide and norfloxacin, while CCCP had no significant effect on YddA-regulated efflux. The results indicated that YddA relies on energy released from ATP hydrolysis to transfer the substrates and YddA is an ABC-type multidrug exporter. Functional study of unknown ATP-binding cassette (ABC) superfamily transporters in the model organism E. coli is conducive to discovering new multidrug resistance-reversal targets and providing references for studying other ABC proteins of unknown function.
|
|
|
KEYWORD
|
|
|
Multidrug efflux gene, yddA, Escherichia coli K-12, ATP-binding cassette, multidrug exporter
|
|
|
FullTexts / Linksout information
|
|
|
|
|
Listed journal information
|
|
  
|