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Generation and characterization of thiol-deficient Mycobacterium tuberculosis mutants
C Sao Emani1,2, M J Williams1, P D Van Helden1
1NRF/DST Centre of Excellence for Biomedical Tuberculosis Research; South African Medical Research Council Centre for Tuberculosis Research; Division of Molecular Biology and Human Genetics, Faculty of Medicine and Health Sciences, Stellenbosch University, Tygerberg 8000, Cape Town, South Africa.
Scientific Data
|September 26, 2018
Summary
Mycothiol (MSH) and ergothioneine (ERG) protect against oxidative stress. This study characterizes M.tb mutants deficient in these thiols, identifying compounds with enhanced activity against them.
Area of Science:
- Microbiology
- Biochemistry
- Drug Discovery
Background:
- Mycothiol (MSH) and ergothioneine (ERG) are crucial thiols for mycobacterial defense against oxidative stress, with potential for mutual compensation.
- Gamma-glutamylcysteine (GGC) also exhibits detoxification properties and is an intermediate in ergothioneine biosynthesis.
- The specific roles of enzymes EgtA-EgtE in ergothioneine production and MshA in mycothiol biosynthesis were previously unclear.
Purpose of the Study:
- To generate and characterize Mycobacterium tuberculosis (M.tb) mutants lacking key enzymes in MSH and ERG biosynthesis.
- To investigate the phenotypic consequences of thiol deficiency in M.tb.
- To identify compounds with differential activity against thiol-deficient M.tb mutants.
Main Methods:
- Generation and phenotypic characterization of M.tb mutants with deletions in genes encoding ergothioneine (EgtA-EgtE) and mycothiol (MshA) biosynthesis enzymes.
- High-throughput screening (HTS) of off-patent drugs and natural compounds against wild-type and thiol-deficient M.tb strains.
- Further investigation into the mode of action of identified compounds.
Main Results:
- Successfully generated and characterized thiol-deficient M.tb mutants.
- Identified several compounds exhibiting enhanced activity against thiol-deficient mutants compared to the wild-type strain.
- Preliminary data on the mode of action of these compounds were obtained.
Conclusions:
- The study provides raw data on thiol biosynthesis mutants in M.tb, offering insights into their physiology.
- Identified compounds represent potential leads for novel anti-mycobacterial strategies targeting thiol-deficient strains.
- Further research is warranted to elucidate the precise mechanisms of identified compounds.
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