Abstract:
Proliferation of Leishmania mexicana promastigotes in synthetic medium can be blocked by the depletion of intracellular polyamine pools induced by the presence of D,L-α-difluoromethylornithine (DFMO), a specific and irreversible inhibitor of ornithine decarboxylase (ODC). Here we report that DEMO-resistant cell lines growing normally at DFMO levels of 10 mM have been obtained from non-proliferating cultures after a single-step selection in the presence of high concentrations of the drug. The DFMO-resistant promastigotes underwent a morphological transformation into an 'amastigote-like' form after incubation for several hours at gradually increasing temperatures up to 35°C. The uptake of DEMO was not significantly altered in the drug-resistant cell lines but in both cases (promastigote and 'amastigote-like' forms) the ODC specific activity was increased approx. 15-fold over the normal enzymic levels found in the wild-type Leishmania. The enzyme affinities for its substrate and for DFMO gave very similar values in the drug-resistant promastigotes and the wild-type parasites. In contrast, ODC from the 'amastigote-like' Leishmania showed a higher affinity for ornithine and a decreased capacity for the binding of DEMO. An 80-fold amplification of the ODC gene and a corresponding increase in its transcripts have been detected in both DEMO-resistant Leishmania cell lines. The drug-resistant phenotypes with their characteristic morphologies, the increased levels of ODC activity and the amplification of the ODC gene have been stable for at least 6 months in the absence of selective pressure.
Registro:
Documento: |
Artículo
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Título: | α-Difluoromethylornithine-resistant cell lines obtained after one-step selection of Leishmania mexicana promastigote cultures |
Autor: | Sánchez, C.P.; Mucci, J.; González, N.S.; Ochoa, A.; Zakin, M.M.; Algranati, I.D. |
Filiación: | Institute de Investigaciones Bioquímicas Fundación Campomar, Facultad de Ciencias Exactas y Naturales, University of Buenos Aires, Patricias Argentinas 435, 1405 Buenos Aires, Argentina Unité d'Expression des Gènes Eucaryotes, Institut Pasteur, Paris, France Zentrum für Infektionsforschung, Universität Würzburg, Röntgenring 11, D-97070 Würzburg, Germany
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Palabras clave: | antitrypanosomal agent; eflornithine; messenger RNA; ornithine; ornithine decarboxylase; ornithine decarboxylase inhibitor; article; binding affinity; cell culture; cell selection; controlled study; drug resistance; drug uptake; enzyme activity; enzyme inhibitor interaction; enzyme specificity; gene amplification; Leishmania mexicana; nonhuman; priority journal; promastigote; Leishmania mexicana |
Año: | 1997
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Volumen: | 324
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Número: | 3
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Página de inicio: | 847
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Página de fin: | 853
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DOI: |
http://dx.doi.org/10.1042/bj3240847 |
Título revista: | Biochemical Journal
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Título revista abreviado: | BIOCHEM. J.
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ISSN: | 02646021
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CODEN: | BIJOA
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CAS: | eflornithine, 67037-37-0, 70052-12-9; ornithine, 70-26-8, 7006-33-9; ornithine decarboxylase, 9024-60-6
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Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v324_n3_p847_Sanchez |
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Citas:
---------- APA ----------
Sánchez, C.P., Mucci, J., González, N.S., Ochoa, A., Zakin, M.M. & Algranati, I.D.
(1997)
. α-Difluoromethylornithine-resistant cell lines obtained after one-step selection of Leishmania mexicana promastigote cultures. Biochemical Journal, 324(3), 847-853.
http://dx.doi.org/10.1042/bj3240847---------- CHICAGO ----------
Sánchez, C.P., Mucci, J., González, N.S., Ochoa, A., Zakin, M.M., Algranati, I.D.
"α-Difluoromethylornithine-resistant cell lines obtained after one-step selection of Leishmania mexicana promastigote cultures"
. Biochemical Journal 324, no. 3
(1997) : 847-853.
http://dx.doi.org/10.1042/bj3240847---------- MLA ----------
Sánchez, C.P., Mucci, J., González, N.S., Ochoa, A., Zakin, M.M., Algranati, I.D.
"α-Difluoromethylornithine-resistant cell lines obtained after one-step selection of Leishmania mexicana promastigote cultures"
. Biochemical Journal, vol. 324, no. 3, 1997, pp. 847-853.
http://dx.doi.org/10.1042/bj3240847---------- VANCOUVER ----------
Sánchez, C.P., Mucci, J., González, N.S., Ochoa, A., Zakin, M.M., Algranati, I.D. α-Difluoromethylornithine-resistant cell lines obtained after one-step selection of Leishmania mexicana promastigote cultures. BIOCHEM. J. 1997;324(3):847-853.
http://dx.doi.org/10.1042/bj3240847