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Infect Immun. Apr 1997; 65(4): 1395–1401.
PMCID: PMC175145

Effects of overexpression of the alkyl hydroperoxide reductase AhpC on the virulence and isoniazid resistance of Mycobacterium tuberculosis.

Abstract

Mutations to the regulatory region of the ahpC gene, resulting in overproduction of alkyl hydroperoxide reductase, were encountered frequently in a large collection of isoniazid (INH)-resistant clinical isolates of Mycobacterium tuberculosis but not in INH-susceptible strains. Overexpression of ahpC did not seem to be important for INH resistance, however, as most of these strains were already defective for catalase-peroxidase, KatG, the enzyme required for activation of INH. Transformation of the INH-susceptible reference strain, M. tuberculosis H37Rv, with plasmids bearing the ahpC genes of M. tuberculosis or M. leprae did not result in a significant increase in the MIC. Two highly INH-resistant mutants of H37Rv, BH3 and BH8, were isolated in vitro and shown to produce no or little KatG activity and, in the case of BH3, to overproduce alkyl hydroperoxide reductase as the result of an ahpC regulatory mutation that was also found in some clinical isolates. The virulence of H37Rv, BH3, and BH8 was studied intensively in three mouse models: fully immunocompetent BALB/c and Black 6 mice, BALB/c major histocompatibility complex class II-knockout mice with abnormally low levels of CD4 T cells and athymic mice producing no cellular immune response. The results indicated that M. tuberculosis strains producing catalase-peroxidase were considerably more virulent in immunocompetent mice than the isogenic KatG-deficient mutants but that loss of catalase-peroxidase was less important when immunodeficient mice, unable to produce activated macrophages, were infected. Restoration of virulence was not seen in an INH-resistant M. tuberculosis strain that overexpressed ahpC, and this finding was confirmed by experiments performed with appropriate M. bovis strains in guinea pigs. Thus, in contrast to catalase-peroxidase, alkyl hydroperoxide reductase does not appear to act as a virulence factor in rodent infections or to play a direct role in INH resistance, although it may be important in maintaining peroxide homeostasis of the organism when KatG activity is low or absent.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Banerjee A, Dubnau E, Quemard A, Balasubramanian V, Um KS, Wilson T, Collins D, de Lisle G, Jacobs WR., Jr inhA, a gene encoding a target for isoniazid and ethionamide in Mycobacterium tuberculosis. Science. 1994 Jan 14;263(5144):227–230. [PubMed]
  • Barletta RG, Kim DD, Snapper SB, Bloom BR, Jacobs WR., Jr Identification of expression signals of the mycobacteriophages Bxb1, L1 and TM4 using the Escherichia-Mycobacterium shuttle plasmids pYUB75 and pYUB76 designed to create translational fusions to the lacZ gene. J Gen Microbiol. 1992 Jan;138(1):23–30. [PubMed]
  • Cole ST. Mycobacterium tuberculosis: drug-resistance mechanisms. Trends Microbiol. 1994 Oct;2(10):411–415. [PubMed]
  • Cosgrove D, Gray D, Dierich A, Kaufman J, Lemeur M, Benoist C, Mathis D. Mice lacking MHC class II molecules. Cell. 1991 Sep 6;66(5):1051–1066. [PubMed]
  • Deretic V, Philipp W, Dhandayuthapani S, Mudd MH, Curcic R, Garbe T, Heym B, Via LE, Cole ST. Mycobacterium tuberculosis is a natural mutant with an inactivated oxidative-stress regulatory gene: implications for sensitivity to isoniazid. Mol Microbiol. 1995 Sep;17(5):889–900. [PubMed]
  • Dessen A, Quémard A, Blanchard JS, Jacobs WR, Jr, Sacchettini JC. Crystal structure and function of the isoniazid target of Mycobacterium tuberculosis. Science. 1995 Mar 17;267(5204):1638–1641. [PubMed]
  • Dhandayuthapani S, Zhang Y, Mudd MH, Deretic V. Oxidative stress response and its role in sensitivity to isoniazid in mycobacteria: characterization and inducibility of ahpC by peroxides in Mycobacterium smegmatis and lack of expression in M. aurum and M. tuberculosis. J Bacteriol. 1996 Jun;178(12):3641–3649. [PMC free article] [PubMed]
  • Dooley SW, Jarvis WR, Martone WJ, Snider DE., Jr Multidrug-resistant tuberculosis. Ann Intern Med. 1992 Aug 1;117(3):257–259. [PubMed]
  • Farr SB, Kogoma T. Oxidative stress responses in Escherichia coli and Salmonella typhimurium. Microbiol Rev. 1991 Dec;55(4):561–585. [PMC free article] [PubMed]
  • Heym B, Alzari PM, Honoré N, Cole ST. Missense mutations in the catalase-peroxidase gene, katG, are associated with isoniazid resistance in Mycobacterium tuberculosis. Mol Microbiol. 1995 Jan;15(2):235–245. [PubMed]
  • Heym B, Cole ST. Isolation and characterization of isoniazid-resistant mutants of Mycobacterium smegmatis and M. aurum. Res Microbiol. 1992 Sep;143(7):721–730. [PubMed]
  • Heym B, Honoré N, Truffot-Pernot C, Banerjee A, Schurra C, Jacobs WR, Jr, van Embden JD, Grosset JH, Cole ST. Implications of multidrug resistance for the future of short-course chemotherapy of tuberculosis: a molecular study. Lancet. 1994 Jul 30;344(8918):293–298. [PubMed]
  • Heym B, Zhang Y, Poulet S, Young D, Cole ST. Characterization of the katG gene encoding a catalase-peroxidase required for the isoniazid susceptibility of Mycobacterium tuberculosis. J Bacteriol. 1993 Jul;175(13):4255–4259. [PMC free article] [PubMed]
  • Kapur V, Li LL, Hamrick MR, Plikaytis BB, Shinnick TM, Telenti A, Jacobs WR, Jr, Banerjee A, Cole S, Yuen KY, et al. Rapid Mycobacterium species assignment and unambiguous identification of mutations associated with antimicrobial resistance in Mycobacterium tuberculosis by automated DNA sequencing. Arch Pathol Lab Med. 1995 Feb;119(2):131–138. [PubMed]
  • Levin ME, Hatfull GF. Mycobacterium smegmatis RNA polymerase: DNA supercoiling, action of rifampicin and mechanism of rifampicin resistance. Mol Microbiol. 1993 Apr;8(2):277–285. [PubMed]
  • Loewen PC, Switala J, Smolenski M, Triggs-Raine BL. Molecular characterization of three mutations in katG affecting the activity of hydroperoxidase I of Escherichia coli. Biochem Cell Biol. 1990 Jul-Aug;68(7-8):1037–1044. [PubMed]
  • MIDDLEBROOK G, COHN ML. Some observations on the pathogenicity of isoniazid-resistant variants of tubercle bacilli. Science. 1953 Sep 11;118(3063):297–299. [PubMed]
  • Morris S, Bai GH, Suffys P, Portillo-Gomez L, Fairchok M, Rouse D. Molecular mechanisms of multiple drug resistance in clinical isolates of Mycobacterium tuberculosis. J Infect Dis. 1995 Apr;171(4):954–960. [PubMed]
  • Musser JM, Kapur V, Williams DL, Kreiswirth BN, van Soolingen D, van Embden JD. Characterization of the catalase-peroxidase gene (katG) and inhA locus in isoniazid-resistant and -susceptible strains of Mycobacterium tuberculosis by automated DNA sequencing: restricted array of mutations associated with drug resistance. J Infect Dis. 1996 Jan;173(1):196–202. [PubMed]
  • Nathan CF, Root RK. Hydrogen peroxide release from mouse peritoneal macrophages: dependence on sequential activation and triggering. J Exp Med. 1977 Dec 1;146(6):1648–1662. [PMC free article] [PubMed]
  • Ordway DJ, Sonnenberg MG, Donahue SA, Belisle JT, Orme IM. Drug-resistant strains of Mycobacterium tuberculosis exhibit a range of virulence for mice. Infect Immun. 1995 Feb;63(2):741–743. [PMC free article] [PubMed]
  • Orme IM. The kinetics of emergence and loss of mediator T lymphocytes acquired in response to infection with Mycobacterium tuberculosis. J Immunol. 1987 Jan 1;138(1):293–298. [PubMed]
  • Rosner JL. Susceptibilities of oxyR regulon mutants of Escherichia coli and Salmonella typhimurium to isoniazid. Antimicrob Agents Chemother. 1993 Oct;37(10):2251–2253. [PMC free article] [PubMed]
  • Rosner JL, Storz G. Effects of peroxides on susceptibilities of Escherichia coli and Mycobacterium smegmatis to isoniazid. Antimicrob Agents Chemother. 1994 Aug;38(8):1829–1833. [PMC free article] [PubMed]
  • Rouse DA, Li Z, Bai GH, Morris SL. Characterization of the katG and inhA genes of isoniazid-resistant clinical isolates of Mycobacterium tuberculosis. Antimicrob Agents Chemother. 1995 Nov;39(11):2472–2477. [PMC free article] [PubMed]
  • Sherman DR, Mdluli K, Hickey MJ, Arain TM, Morris SL, Barry CE, 3rd, Stover CK. Compensatory ahpC gene expression in isoniazid-resistant Mycobacterium tuberculosis. Science. 1996 Jun 14;272(5268):1641–1643. [PubMed]
  • Sherman DR, Sabo PJ, Hickey MJ, Arain TM, Mahairas GG, Yuan Y, Barry CE, 3rd, Stover CK. Disparate responses to oxidative stress in saprophytic and pathogenic mycobacteria. Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6625–6629. [PMC free article] [PubMed]
  • Snapper SB, Melton RE, Mustafa S, Kieser T, Jacobs WR., Jr Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis. Mol Microbiol. 1990 Nov;4(11):1911–1919. [PubMed]
  • Storz G, Tartaglia LA, Ames BN. Transcriptional regulator of oxidative stress-inducible genes: direct activation by oxidation. Science. 1990 Apr 13;248(4952):189–194. [PubMed]
  • Telenti A, Honoré N, Bernasconi C, March J, Ortega A, Heym B, Takiff HE, Cole ST. Genotypic assessment of isoniazid and rifampin resistance in Mycobacterium tuberculosis: a blind study at reference laboratory level. J Clin Microbiol. 1997 Mar;35(3):719–723. [PMC free article] [PubMed]
  • Wilson TM, Collins DM. ahpC, a gene involved in isoniazid resistance of the Mycobacterium tuberculosis complex. Mol Microbiol. 1996 Mar;19(5):1025–1034. [PubMed]
  • Wilson TM, de Lisle GW, Collins DM. Effect of inhA and katG on isoniazid resistance and virulence of Mycobacterium bovis. Mol Microbiol. 1995 Mar;15(6):1009–1015. [PubMed]
  • Zhang Y, Heym B, Allen B, Young D, Cole S. The catalase-peroxidase gene and isoniazid resistance of Mycobacterium tuberculosis. Nature. 1992 Aug 13;358(6387):591–593. [PubMed]

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