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Abstract:

Trypanosoma cruzi, the etiologic agent of Chagas' disease, is a polyamine auxotroph organism because its genome contains neither ornithine decarboxylase (ODC) nor arginine decarboxylase (ADC) genes, presumably lost during evolution. After transformation with a recombinant plasmid bearing the complete coding region of Crithidia fasciculata ODC gene, the transgenic parasites were able to synthesize putrescine and simultaneously became susceptible to α-difluoromethylornithine (DFMO), an irreversible inhibitor of ODC. We have studied the emergence of DFMO-resistant T. cruzi after one-step selection of ODC-transformed parasites cultivated in the presence of high levels of the drug (5 mM). Our results have indicated a duplication of the ODC gene copy number in the drug-resistant cell line. The ODC transcripts and the corresponding translation products showed very significant increases (about 7- and 25-fold, respectively) in DFMO-resistant parasites, while the ODC enzymatic activity was 5 times higher than in drug-sensitive T. cruzi. The unequal increases of ODC protein and enzymatic activity in DFMO-resistant protozoa strongly suggest that in addition to gene amplification and enhanced transcription and translation, the assembly of ODC polypeptide chains into dimeric active enzyme molecules might also contribute to regulate the development of DFMO resistance. © 2007 Elsevier B.V. All rights reserved.

Registro:

Documento: Artículo
Título:Trypanosoma cruzi as a model system to study the expression of exogenous genes coding for polyamine biosynthetic enzymes. Induction of DFMO resistance in transgenic parasites
Autor:Carrillo, C.; González, N.S.; Algranati, I.D.
Filiación:Fundación Instituto Leloir, IIBBA-CONICET, Facultad de Ciencias Exactas y Naturales, C1405BWE Buenos Aires, Argentina
Palabras clave:DFMO resistance; Ornithine decarboxylase; Polyamine biosynthesis; Transgenic parasites; Trypanosoma cruzi; eflornithine; messenger RNA; ornithine decarboxylase; polyamine; article; controlled study; enzyme activity; gene amplification; gene duplication; gene expression; genetic transcription; nonhuman; priority journal; RNA translation; transgenic organism; Trypanosoma cruzi; Animals; Animals, Genetically Modified; Base Sequence; Biogenic Polyamines; Blotting, Northern; Blotting, Southern; DNA Primers; Eflornithine; Gene Expression; Ornithine Decarboxylase; Polymerase Chain Reaction; Trypanosoma cruzi; Crithidia fasciculata; Protozoa; Trypanosoma cruzi
Año:2007
Volumen:1770
Número:12
Página de inicio:1605
Página de fin:1611
DOI: http://dx.doi.org/10.1016/j.bbagen.2007.08.013
Título revista:Biochimica et Biophysica Acta - General Subjects
Título revista abreviado:Biochim. Biophys. Acta Gen. Subj.
ISSN:03044165
CODEN:BBGSB
CAS:eflornithine, 67037-37-0, 70052-12-9; ornithine decarboxylase, 9024-60-6; Biogenic Polyamines; DNA Primers; Eflornithine, 70052-12-9; Ornithine Decarboxylase, EC 4.1.1.17
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03044165_v1770_n12_p1605_Carrillo

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Citas:

---------- APA ----------
Carrillo, C., González, N.S. & Algranati, I.D. (2007) . Trypanosoma cruzi as a model system to study the expression of exogenous genes coding for polyamine biosynthetic enzymes. Induction of DFMO resistance in transgenic parasites. Biochimica et Biophysica Acta - General Subjects, 1770(12), 1605-1611.
http://dx.doi.org/10.1016/j.bbagen.2007.08.013
---------- CHICAGO ----------
Carrillo, C., González, N.S., Algranati, I.D. "Trypanosoma cruzi as a model system to study the expression of exogenous genes coding for polyamine biosynthetic enzymes. Induction of DFMO resistance in transgenic parasites" . Biochimica et Biophysica Acta - General Subjects 1770, no. 12 (2007) : 1605-1611.
http://dx.doi.org/10.1016/j.bbagen.2007.08.013
---------- MLA ----------
Carrillo, C., González, N.S., Algranati, I.D. "Trypanosoma cruzi as a model system to study the expression of exogenous genes coding for polyamine biosynthetic enzymes. Induction of DFMO resistance in transgenic parasites" . Biochimica et Biophysica Acta - General Subjects, vol. 1770, no. 12, 2007, pp. 1605-1611.
http://dx.doi.org/10.1016/j.bbagen.2007.08.013
---------- VANCOUVER ----------
Carrillo, C., González, N.S., Algranati, I.D. Trypanosoma cruzi as a model system to study the expression of exogenous genes coding for polyamine biosynthetic enzymes. Induction of DFMO resistance in transgenic parasites. Biochim. Biophys. Acta Gen. Subj. 2007;1770(12):1605-1611.
http://dx.doi.org/10.1016/j.bbagen.2007.08.013