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Molecular cloning and functional characterization of an ATP-binding cassette transporter OtrC from Streptomyces rimosus
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Molecular cloning and functional characterization of an ATP-binding cassette transporter OtrC from Streptomyces rimosus
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Journal Article
Molecular cloning and functional characterization of an ATP-binding cassette transporter OtrC from Streptomyces rimosus
2012
Overview
Background
The
otrC
gene of
Streptomyces rimosus
was previously annotated as an oxytetracycline (OTC) resistance protein. However, the amino acid sequence analysis of OtrC shows that it is a putative ATP-binding cassette (ABC) transporter with multidrug resistance function. To our knowledge, none of the ABC transporters in
S. rimosus
have yet been characterized. In this study, we aimed to characterize the multidrug exporter function of OtrC and evaluate its relevancy to OTC production.
Results
In order to investigate OtrC’s function,
otrC
is cloned and expressed in
E. coli
The exporter function of OtrC was identified by ATPase activity determination and ethidium bromide efflux assays. Also, the susceptibilities of OtrC-overexpressing cells to several structurally unrelated drugs were compared with those of OtrC-non-expressing cells by minimal inhibitory concentration (MIC) assays, indicating that OtrC functions as a drug exporter with a broad range of drug specificities. The OTC production was enhanced by 1.6-fold in M4018 (
P
= 0.000877) and 1.4-fold in SR16 (
P
= 0.00973) duplication mutants, while it decreased to 80% in disruption mutants (
P
= 0.0182 and 0.0124 in M4018 and SR16, respectively).
Conclusions
The results suggest that OtrC is an ABC transporter with multidrug resistance function, and plays an important role in self-protection by drug efflux mechanisms. This is the first report of such a protein in
S. rimosus
, and
otrC
could be a valuable target for genetic manipulation to improve the production of industrial antibiotics.
Publisher
BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
Subject
/ Adenosine triphosphatase genes
/ Analysis
/ ATP
/ ATP-binding cassette transporter
/ ATP-Binding Cassette Transporters - chemistry
/ ATP-Binding Cassette Transporters - genetics
/ ATP-Binding Cassette Transporters - metabolism
/ Bacterial Proteins - chemistry
/ Bacterial Proteins - genetics
/ Bacterial Proteins - metabolism
/ Biomedical Engineering/Biotechnology
/ Chemistry and Materials Science
/ Cloning
/ Drug resistance in microorganisms
/ E coli
/ Escherichia coli - metabolism
/ Genes
/ Mutation
/ OtrC
/ Oxytetracycline - biosynthesis
/ Plant Breeding/Biotechnology
/ Proteins
/ Recombinant Proteins - biosynthesis
/ Recombinant Proteins - chemistry
