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

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 co...

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Autores principales:	Carrillo, C., González, N.S., Algranati, I.D.
Formato:	JOUR
Materias:	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 Animals Animals, Genetically Modified Base Sequence Biogenic Polyamines Blotting, Northern Blotting, Southern DNA Primers Eflornithine Gene Expression Ornithine Decarboxylase Polymerase Chain Reaction Crithidia fasciculata Protozoa
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_03044165_v1770_n12_p1605_Carrillo
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_03044165_v1770_n12_p1605_Carrillo
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spelling	todo:paper_03044165_v1770_n12_p1605_Carrillo2023-10-03T15:20:43Z 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 Carrillo, C. González, N.S. Algranati, I.D. 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 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. Fil:Carrillo, C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:González, N.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Algranati, I.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_03044165_v1770_n12_p1605_Carrillo
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	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
spellingShingle	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 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
topic_facet	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
description	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.
format	JOUR
author	Carrillo, C. González, N.S. Algranati, I.D.
author_facet	Carrillo, C. González, N.S. Algranati, I.D.
author_sort	Carrillo, C.
title	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
title_short	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
title_full	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
title_fullStr	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
title_full_unstemmed	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
title_sort	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
url	http://hdl.handle.net/20.500.12110/paper_03044165_v1770_n12_p1605_Carrillo
work_keys_str_mv	AT carrilloc trypanosomacruziasamodelsystemtostudytheexpressionofexogenousgenescodingforpolyaminebiosyntheticenzymesinductionofdfmoresistanceintransgenicparasites AT gonzalezns trypanosomacruziasamodelsystemtostudytheexpressionofexogenousgenescodingforpolyaminebiosyntheticenzymesinductionofdfmoresistanceintransgenicparasites AT algranatiid trypanosomacruziasamodelsystemtostudytheexpressionofexogenousgenescodingforpolyaminebiosyntheticenzymesinductionofdfmoresistanceintransgenicparasites
_version_	1782030797065158656

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

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