Evidence that trypanothione reductase is an essential enzyme in Leishmania by targeted replacement of the tryA gene locus.

Abstract:

:Trypanothione reductase (TR), a flavoprotein oxidoreductase central to the unique thiol-redox system that operates in trypanosomatid protozoa, has been proposed as a potential target for the chemotherapy of trypanosomatid infections. In this study, targeted gene replacement was used to obtain evidence that TR is an essential cellular component and that its physiological function is crucial for parasite survival. Precise replacement of the Leishmania donovani tryA gene encoding TR was only possible upon simultaneous expression of the tryA coding region from an episome; in its absence, attempted removal of the last tryA allele invariably led to the generation of an extra copy of tryA, seemingly as a result of selective chromosomal polysomy. Partial replacement mutants were drastically affected in their ability to survive inside cytokine-activated macrophages in a murine model of Leishmania infection. As no compensatory mechanism for the partial loss of TR activity was observed in these mutants and as it was not possible to obtain viable Leishmania devoid of TR catalytic activity, specific inhibitors of this enzyme are likely to be useful anti-leishmanial agents for chemotherapeutic use.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Tovar J,Wilkinson S,Mottram JC,Fairlamb AH

doi

10.1046/j.1365-2958.1998.00968.x

subject

Has Abstract

pub_date

1998-07-01 00:00:00

pages

653-60

issue

2

eissn

0950-382X

issn

1365-2958

journal_volume

29

pub_type

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