Abstract |
Capreomycin (CAP) is an important second-line drug for multidrug-resistant tuberculosis. To further define the drug resistance mechanism of CAP, a Mycobacterium smegmatis transposon mutant library was constructed using Tn5 transposon for screening isolates with enhanced CAP resistance. A mutant (named C4) with fourfold increased CAP resistance was isolated and characterized. Tn5 was found to be inserted into MSMEG_0841, an annotated pseudogene. However, knockout demonstrated that MSMEG_0841 was not responsible for CAP resistance. We further sequenced the whole genome of C4 and found an A to G substitution in the overlap region between tlyA and ppnK, which leads a stop codon mutation in upstream tlyA and a T2A mutation in downstream ppnK. Mutation in the overlap might confer the dysfuction of both genes. tlyA is a known gene involved in CAP action. Overexpression of ppnK in both Escherichia coli and M. smegmatis confer subtle susceptible to CAP. Taken together, our study found that a novel mutation involved in CAP resistance.
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Authors | Qinglin Du, Quanxin Long, Jinxiao Mao, Tiwei Fu, Xiangke Duan, Jianping Xie |
Journal | IUBMB life
(IUBMB Life)
Vol. 66
Issue 6
Pg. 405-14
(Jun 2014)
ISSN: 1521-6551 [Electronic] England |
PMID | 24890219
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | © 2014 International Union of Biochemistry and Molecular Biology. |
Chemical References |
- Bacterial Proteins
- Oligonucleotides
- TlyA protein, Mycobacterium tuberculosis
- Capreomycin
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Topics |
- Bacterial Proteins
(genetics)
- Base Sequence
- Capreomycin
- Drug Resistance, Bacterial
(genetics)
- Gene Knockout Techniques
- Genome, Bacterial
(genetics)
- Microscopy, Electron, Scanning
- Molecular Sequence Data
- Mutation, Missense
(genetics)
- Mycobacterium
(genetics)
- Oligonucleotides
(genetics)
- Real-Time Polymerase Chain Reaction
- Reverse Transcriptase Polymerase Chain Reaction
- Sequence Analysis, DNA
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