Abstract
BackgroundMycobacterium tuberculosis resides inside host macrophages during infection and adapts to resilient stresses generated by the host immune system. As a response, M. tuberculosis codes for short-chain dehydrogenases/reductases (SDRs). These SDRs are nicotinamide adenine dinucleotide-reliant oxidoreductases involved in cell homeostasis. The precise function of oxidoreductases in bacteria especially M. tuberculosis were not fully explored. This study aimed to know the detail functional role of one of the oxidoreductase Rv0148 in M. tuberculosis.ResultsIn silico analysis revealed that Rv0148 interacts with Htdy (Rv3389) and the protein interactions were confirmed using far western blot. Gene knockout mutant of Rv0148 in M. tuberculosis was constructed by specialized transduction. Macrophage cell line infection with this knockout mutant showed increased expression of pro-inflammatory cytokines. This knockout mutant is sensitive to oxidative, nitrogen, redox and electron transport inhibitor stress agents. Drug susceptibility testing of the deletion mutant showed resistance to first-line drugs such as streptomycin and ethambutol and second-line aminoglycosides such as amikacin and kanamycin. Based on interactorme analysis for Rv0148 using STRING database, we identified 220 most probable interacting partners for Htdy protein. In the Rv0148 knockout mutants, high expression of htdy was observed and we hypothesize that this would have perturbed the interactome thus resulting in drug resistance. Finally, we propose that Rv0148 and Htdy are functionally interconnected and involved in drug resistance and cell homeostasis of M. tuberculosis.ConclusionsOur study suggests that Rv0148 plays a significant role in various functional aspects such as intermediatory metabolism, stress, homeostasis and also in drug resistance.
Highlights
Mycobacterium tuberculosis resides inside host macrophages during infection and adapts to resilient stresses generated by the host immune system
By in silico approach we identified that Rv0148 mainly interacts with hydroxyl acyl thioester dehydratase Htdy, a protein interaction which was confirmed by far western blotting (WB) and pull-down assay
Sequence homologues of Rv0148 To identify the sequence homologues of Rv0148 in other organisms, blast search was performed against the nonreductant database in NCBI; 104 sequence homologues were selected and the sequences were reconfirmed based on short dehydrogenase domain in these sequences
Summary
Mycobacterium tuberculosis resides inside host macrophages during infection and adapts to resilient stresses generated by the host immune system. M. tuberculosis codes for short-chain dehydrogenases/ reductases (SDRs). These SDRs are nicotinamide adenine dinucleotide-reliant oxidoreductases involved in cell homeostasis. The precise function of oxidoreductases in bacteria especially M. tuberculosis were not fully explored. This study aimed to know the detail functional role of one of the oxidoreductase Rv0148 in M. tuberculosis. Tuberculosis (TB) remains one of the world’s leading cause of death every year among infectious diseases [1]. Due to the absence of oxyR in M. tuberculosis, peroxidase stress is managed by alkylhydroperoxidase reductase (AhpC) by detoxifying the peroxides [12]. Though the components involved in the oxidative stress of M. tuberculosis have been identified, the functional importance remains unclear. Gene interaction and knockout studies thoroughly predict the functional interconnection between the genes and their role in the metabolism of bacteria
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