S-adenosylmethionine decarboxylase from Leishmania donovani: Molecular, genetic, and biochemical characterization of null mutants and overproducers

Sigrid C. Roberts, Jerry Scott, Judith E. Gasteier, Yuqui Jiang, Benjamin Brooks, Armando Jardim, Nicola Carter, Olle Heby, Buddy Ullman

Research output: Contribution to journalArticle

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Abstract

The polyamine biosynthetic enzyme, S-adenosylmethionine decarboxylase (ADOMETDC) has been advanced as a potential target for antiparasitic chemotherapy. To investigate the importance of this protein in a model parasite, the gene encoding ADOMETDC has been cloned and sequenced from Leishmania donovani. The Δadometdc null mutants were created in the insect vector form of the parasite by double targeted gene replacement. The Δadometdc strains were incapable of growth in medium without polyamines; however, auxotrophy could be rescued by spermidine but not by putrescine, spermine, or methylthioadenosine. Incubation of Δadometdc parasites in medium lacking polyamines resulted in a drastic increase of putrescine and glutathione levels with a concomitant decrease in the amounts of spermidine and the spermidine-containing thiol trypanothione. Parasites transfected with an episomal ADOMETDC construct were created in both wild type and Δadometdc parasites. ADOMETDC overexpression abrogated polyamine auxotrophy in the Δadometdc L. donovani. In addition, ADOMETDC overproduction in wild type parasites alleviated the toxic effects of 5′-(((Z)-4-amino-2-butenyl)methylamino)-5′-deoxyadenosine (MDL 73811), but not pentamidine, berenil, or methyl-glyoxyl bis(guanylhydrazone), all inhibitors of ADOMETDC activities in vitro. The molecular, biochemical, and genetic characterization of ADOMETDC establishes that it is essential in L. donovani promastigotes and a potential target for therapeutic validation.

Original languageEnglish (US)
Pages (from-to)5902-5909
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number8
DOIs
StatePublished - Feb 22 2002

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Adenosylmethionine Decarboxylase
Leishmania donovani
Molecular Biology
Parasites
Polyamines
Spermidine
Putrescine
Insect Vectors
Pentamidine
Antiparasitic Agents
Gene encoding
Chemotherapy
Spermine
Poisons
Sulfhydryl Compounds
Genes
Glutathione
Drug Therapy
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

S-adenosylmethionine decarboxylase from Leishmania donovani : Molecular, genetic, and biochemical characterization of null mutants and overproducers. / Roberts, Sigrid C.; Scott, Jerry; Gasteier, Judith E.; Jiang, Yuqui; Brooks, Benjamin; Jardim, Armando; Carter, Nicola; Heby, Olle; Ullman, Buddy.

In: Journal of Biological Chemistry, Vol. 277, No. 8, 22.02.2002, p. 5902-5909.

Research output: Contribution to journalArticle

Roberts, SC, Scott, J, Gasteier, JE, Jiang, Y, Brooks, B, Jardim, A, Carter, N, Heby, O & Ullman, B 2002, 'S-adenosylmethionine decarboxylase from Leishmania donovani: Molecular, genetic, and biochemical characterization of null mutants and overproducers', Journal of Biological Chemistry, vol. 277, no. 8, pp. 5902-5909. https://doi.org/10.1074/jbc.M110118200
Roberts, Sigrid C. ; Scott, Jerry ; Gasteier, Judith E. ; Jiang, Yuqui ; Brooks, Benjamin ; Jardim, Armando ; Carter, Nicola ; Heby, Olle ; Ullman, Buddy. / S-adenosylmethionine decarboxylase from Leishmania donovani : Molecular, genetic, and biochemical characterization of null mutants and overproducers. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 8. pp. 5902-5909.
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