Antisense RNA to ahpC, an oxidative stress defence gene involved in isoniazid resistance, indicates that AhpC of Mycobacterium bovis has virulence properties

Theresa Wilson, Geoffrey W. De Lisle, Jovita A. Marcinkeviciene, John S. Blanchard, Desmond M. Collins

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Antisense RNA is a versatile tool for reducing gene expression. It was used to determine if ahpC, a gene that is involved in defence against oxidative stress and isoniazid (INH) resistance, is important for virulence of Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex. Antisense RNA constructs of ahpC were made using different strength promoters in front of a reversed coding sequence of ahpC. These constructs were electroporated into a virulent wild-type M. bovis strain and a moderately virulent INH-resistant M. bovis strain that was catalase/peroxidase-negative. Down-regulation of protein synthesis occurred and this was visualized by immunoblotting. All strains containing antisense RNA were markedly less virulent than their parent strains in guinea pigs. M. bovis with an up-regulated ahpC gene was more resistant to cumene hydroperoxide than its parent strain, which had a wild-type ahpC promoter. These results agree with a model of INH resistance in which overexpression of AhpC compensates in some INH-resistant strains for loss of catalase/peroxidase by maintaining the ability to defend against oxidative stress mediated through organic peroxides. In addition, normal expression of AhpC is crucial for maintaining the virulence of wild-type M. bovis, which has normal catalase/peroxidase levels.

Original languageEnglish (US)
Pages (from-to)2687-2695
Number of pages9
JournalMicrobiology
Volume144
Issue number10
StatePublished - Oct 1998

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Antisense RNA
Isoniazid
Mycobacterium bovis
Virulence
Oxidative Stress
Catalase
Peroxidase
Genes
Peroxides
Mycobacterium tuberculosis
Immunoblotting
Guinea Pigs
Down-Regulation
Gene Expression
Proteins

Keywords

  • AhpC
  • Antisense RNA
  • Mycobacterium tuberculosis complex
  • Oxidative stress
  • Virulence

ASJC Scopus subject areas

  • Microbiology

Cite this

Antisense RNA to ahpC, an oxidative stress defence gene involved in isoniazid resistance, indicates that AhpC of Mycobacterium bovis has virulence properties. / Wilson, Theresa; De Lisle, Geoffrey W.; Marcinkeviciene, Jovita A.; Blanchard, John S.; Collins, Desmond M.

In: Microbiology, Vol. 144, No. 10, 10.1998, p. 2687-2695.

Research output: Contribution to journalArticle

Wilson, Theresa ; De Lisle, Geoffrey W. ; Marcinkeviciene, Jovita A. ; Blanchard, John S. ; Collins, Desmond M. / Antisense RNA to ahpC, an oxidative stress defence gene involved in isoniazid resistance, indicates that AhpC of Mycobacterium bovis has virulence properties. In: Microbiology. 1998 ; Vol. 144, No. 10. pp. 2687-2695.
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