Effect of Oxidative Stress on AhpC Activity and Virulence in katG Ser315 Thr Mycobacterium tuberculosis Mutant
https://doi.org/10.22146/ijbiotech.16369 Ning Rintiswati(1*), Tri Wibawa(2), Widya Asmara(3), Hardyanto Soebono(4)
(1) Doctoral Program of Health and Medical Science, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia Department of Microbiology, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia
(2) Department of Microbiology, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia
(3) Department of Microbiology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia
(4) Department of Dermathology and Venereology, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia
(*) Corresponding Author
Abstract
Mycobacterium tuberculosis strains resistance to INH is mainly caused by the alteration in several genes
encoding the molecular targets. Mutation of katG at codon 315 especially Ser315Thr are responsible for
INH resistance in a large proportion of TB cases. The aim of this study is to evaluate the influence of stress
oxidative on AhpC activity of katG Ser315Thr of M.tuberculosis, and to find out the relation of AhpC and the
virulence of this mutant. The study design was laboratoric experimental, subjects of study were M.tuberculosis
INH resistance strains, and the treatment were serial dose of H2O2. Eighty five M.tuberculosis INH resistant
clinical strain were screened for mutation of katGSer315Thr by PCR/RFLP and characterized on the basis of
phenotypic properties (catalase activity and AhpC activity). AhpC activity of katG Ser315Thr M.tuberculosis
strains in response to oxidative stress condition was evaluated by culturing the strains on liquid culture
medium containing 1mM H2O2.To ascertain role of AhpC in the virulence of katGSer315Thr mutant strains, the
mutants were infected into human macrophages culture, and several indicator of virulence were observed (i.e:
replication competence, and apoptosis induction on human macrophages). The results showed that katG Ser
315Thr were identified in 23 (27,05%) of 85 INH resistance strains, all mutant strains had decrease of catalase
activity. AhpC activity of katG Ser315Thr of M.tuberculosis increased significantly with increase of hydrogen
peroxide dose. In addition , it has been shown that increased AhpC activity related to replication ability of
mutant, and reduction of apoptosis macrophages induction significantly. We conclude that the production
of AhpC of katG Ser315Thr M.tuberculosis induced by oxidative stress. There was a role of AhpC in virulence
of the M.tuberculosis katG Ser315Thr strains by replication capability and macrophages apoptosis.
encoding the molecular targets. Mutation of katG at codon 315 especially Ser315Thr are responsible for
INH resistance in a large proportion of TB cases. The aim of this study is to evaluate the influence of stress
oxidative on AhpC activity of katG Ser315Thr of M.tuberculosis, and to find out the relation of AhpC and the
virulence of this mutant. The study design was laboratoric experimental, subjects of study were M.tuberculosis
INH resistance strains, and the treatment were serial dose of H2O2. Eighty five M.tuberculosis INH resistant
clinical strain were screened for mutation of katGSer315Thr by PCR/RFLP and characterized on the basis of
phenotypic properties (catalase activity and AhpC activity). AhpC activity of katG Ser315Thr M.tuberculosis
strains in response to oxidative stress condition was evaluated by culturing the strains on liquid culture
medium containing 1mM H2O2.To ascertain role of AhpC in the virulence of katGSer315Thr mutant strains, the
mutants were infected into human macrophages culture, and several indicator of virulence were observed (i.e:
replication competence, and apoptosis induction on human macrophages). The results showed that katG Ser
315Thr were identified in 23 (27,05%) of 85 INH resistance strains, all mutant strains had decrease of catalase
activity. AhpC activity of katG Ser315Thr of M.tuberculosis increased significantly with increase of hydrogen
peroxide dose. In addition , it has been shown that increased AhpC activity related to replication ability of
mutant, and reduction of apoptosis macrophages induction significantly. We conclude that the production
of AhpC of katG Ser315Thr M.tuberculosis induced by oxidative stress. There was a role of AhpC in virulence
of the M.tuberculosis katG Ser315Thr strains by replication capability and macrophages apoptosis.
Keywords
katG Ser315Thr Mycobacterium tuberculosis; oxidative stress; AhpC; virulence
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PDF DOI: https://doi.org/10.22146/ijbiotech.16369
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