Protective role of the Mycobacterium smegmatis IdeR against reactive oxygen species and isoniazid toxicity.
OBJECTIVE
To understand the mechanism by which IdeR is necessary for maintaining wild type levels of KatG and SodA enzyme activity and normal isoniazid (INH) resistance.


METHODS
To identify the step(s) of SodA and KatG function that were affected by the ideR mutation, quantitative western immunoassays and ribonucleic acid (RNA) hybridizations were performed. To see if the increased INH sensitivity of the ideR mutant was caused by lower SodA activity, the Mycobacterium smegmatis sod gene was inactivated.


RESULTS
The levels of KatG and SodA mRNA and protein in the M. smegmatis IdeR mutant are decreased to approximately 20-40% of those observed in the wild type parent strain. This is quantitatively similar to the decrease in KatG and SodA enzyme activities originally observed in the ideR strain. The M. smegmatis sodA mutant was slightly more sensitive to INH, compared to the wild type strain and was more resistant than the ideR mutant.


CONCLUSIONS
IdeR is necessary for full expression of the M. smegmatis katG and sodA genes. It is not yet known whether this protein acts directly at the gene level. The lower levels of SodA contribute slightly to the increased susceptibility to INH of the ideR mutant, but cannot explain the magnitude of the INH sensitivity observed when IdeR is not present. These data suggest that IdeR is a regulator of the cellular stress response, as it has a protective role in cells facing environmental stresses, such as increased levels of reactive oxygen species and INH toxic intermediates. These conclusions do not necessarily apply to IdeR's role in M. tuberculosis physiology, since we have not inactivated its gene in this pathogen.
Dussurget
O
O
Department of Microbiology, Public Health Research Institute, New York, NY 10016, USA.
Rodriguez
M
M
Smith
I
I
D000995:Antitubercular Agents; D001426:Bacterial Proteins; D015153:Blotting, Western; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005798:Genes, Bacterial; D007538:Isoniazid; D009154:Mutation; D020102:Mycobacterium smegmatis / Q000201:enzymology / Q000235:genetics*; D010544:Peroxidases / Q000032:analysis; D054464:Peroxiredoxins; D017382:Reactive Oxygen Species; D013482:Superoxide Dismutase / Q000032:analysis
D000995:Antitubercular Agents; D001426:Bacterial Proteins; D017382:Reactive Oxygen Species; D010544:Peroxidases; D054464:Peroxiredoxins; C076136:catalase HPI; D013482:Superoxide Dismutase; D007538:Isoniazid