Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle

Trypanosoma cruzi, etiological agent of Chagas' disease, has a complex life cycle which involves the invasion of mammalian host cells, differentiation and intracellular replication. Here we report the first insights into the biological role of a poly(ADP-ribose) glycohydrolase in a trypanosomat...

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Autores principales: Vilchez Larrea, S.C., Schlesinger, M., Kevorkian, M.L., Flawiá, M.M., Alonso, G.D., Fernández Villamil, S.H.
Formato: Artículo publishedVersion
Lenguaje:Inglés
Publicado: 2013
Materias:
DNA
RNA
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_19326203_v8_n6_p_VilchezLarrea
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spelling paperaa:paper_19326203_v8_n6_p_VilchezLarrea2023-06-12T16:51:48Z Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle PLoS ONE 2013;8(6) Vilchez Larrea, S.C. Schlesinger, M. Kevorkian, M.L. Flawiá, M.M. Alonso, G.D. Fernández Villamil, S.H. antibody cell enzyme DNA glycosidase inhibitor hydroxyurea poly(adenosine diphosphate ribose) poly(adenosine diphosphate ribose) glycohydrolase RNA unclassified drug article catalysis cell cycle progression cell nucleus Chagas disease concentration (parameters) DNA damage electron microscopy enzyme localization epimastigote gene gene expression genetic analysis genetic conservation growth rate host cell human human cell immunofluorescence test in vitro study kinetoplastid life cycle stage Kinetoplastida nonhuman nucleotide sequence RNA interference TcPARG gene Trypanosoma cruzi trypomastigote Vero cell Western blotting Adenosine Diphosphate Animals Blotting, Northern Blotting, Southern Blotting, Western Catalysis Cell Cycle Cell Line, Tumor Cercopithecus aethiops Chagas Disease Fluorescent Antibody Technique, Indirect Glycoside Hydrolases Humans Hydroxyurea Life Cycle Stages Microscopy, Electron Pyrrolidines Trypanosoma cruzi Vero Cells Mammalia Trypanosoma cruzi Trypanosomatidae Trypanosoma cruzi, etiological agent of Chagas' disease, has a complex life cycle which involves the invasion of mammalian host cells, differentiation and intracellular replication. Here we report the first insights into the biological role of a poly(ADP-ribose) glycohydrolase in a trypanosomatid (TcPARG). In silico analysis of the TcPARG gene pointed out the conservation of key residues involved in the catalytic process and, by Western blot, we demonstrated that it is expressed in a life stage-dependant manner. Indirect immunofluorescence assays and electron microscopy using an anti-TcPARG antibody showed that this enzyme is localized in the nucleus independently of the presence of DNA damage or cell cycle stage. The addition of poly(ADP-ribose) glycohydrolase inhibitors ADP-HPD (adenosine diphosphate (hydroxymethyl) pyrrolidinediol) or DEA (6,9-diamino-2-ethoxyacridine lactate monohydrate) to the culture media, both at a 1 μM concentration, reduced in vitro epimastigote growth by 35% and 37% respectively, when compared to control cultures. We also showed that ADP-HPD 1 μM can lead to an alteration in the progression of the cell cycle in hydroxyurea synchronized cultures of T. cruzi epimastigotes. Outstandingly, here we demonstrate that the lack of poly(ADP-ribose) glycohydrolase activity in Vero and A549 host cells, achieved by chemical inhibition or iRNA, produces the reduction of the percentage of infected cells as well as the number of amastigotes per cell and trypomastigotes released, leading to a nearly complete abrogation of the infection process. We conclude that both, T. cruzi and the host, poly(ADP-ribose) glycohydrolase activities are important players in the life cycle of Trypanosoma cruzi, emerging as a promising therapeutic target for the treatment of Chagas' disease. © 2013 Vilchez Larrea et al. Fil:Flawiá, M.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Alonso, G.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2013 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_19326203_v8_n6_p_VilchezLarrea
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language Inglés
orig_language_str_mv eng
topic antibody
cell enzyme
DNA
glycosidase inhibitor
hydroxyurea
poly(adenosine diphosphate ribose)
poly(adenosine diphosphate ribose) glycohydrolase
RNA
unclassified drug
article
catalysis
cell cycle progression
cell nucleus
Chagas disease
concentration (parameters)
DNA damage
electron microscopy
enzyme localization
epimastigote
gene
gene expression
genetic analysis
genetic conservation
growth rate
host cell
human
human cell
immunofluorescence test
in vitro study
kinetoplastid life cycle stage
Kinetoplastida
nonhuman
nucleotide sequence
RNA interference
TcPARG gene
Trypanosoma cruzi
trypomastigote
Vero cell
Western blotting
Adenosine Diphosphate
Animals
Blotting, Northern
Blotting, Southern
Blotting, Western
Catalysis
Cell Cycle
Cell Line, Tumor
Cercopithecus aethiops
Chagas Disease
Fluorescent Antibody Technique, Indirect
Glycoside Hydrolases
Humans
Hydroxyurea
Life Cycle Stages
Microscopy, Electron
Pyrrolidines
Trypanosoma cruzi
Vero Cells
Mammalia
Trypanosoma cruzi
Trypanosomatidae
spellingShingle antibody
cell enzyme
DNA
glycosidase inhibitor
hydroxyurea
poly(adenosine diphosphate ribose)
poly(adenosine diphosphate ribose) glycohydrolase
RNA
unclassified drug
article
catalysis
cell cycle progression
cell nucleus
Chagas disease
concentration (parameters)
DNA damage
electron microscopy
enzyme localization
epimastigote
gene
gene expression
genetic analysis
genetic conservation
growth rate
host cell
human
human cell
immunofluorescence test
in vitro study
kinetoplastid life cycle stage
Kinetoplastida
nonhuman
nucleotide sequence
RNA interference
TcPARG gene
Trypanosoma cruzi
trypomastigote
Vero cell
Western blotting
Adenosine Diphosphate
Animals
Blotting, Northern
Blotting, Southern
Blotting, Western
Catalysis
Cell Cycle
Cell Line, Tumor
Cercopithecus aethiops
Chagas Disease
Fluorescent Antibody Technique, Indirect
Glycoside Hydrolases
Humans
Hydroxyurea
Life Cycle Stages
Microscopy, Electron
Pyrrolidines
Trypanosoma cruzi
Vero Cells
Mammalia
Trypanosoma cruzi
Trypanosomatidae
Vilchez Larrea, S.C.
Schlesinger, M.
Kevorkian, M.L.
Flawiá, M.M.
Alonso, G.D.
Fernández Villamil, S.H.
Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle
topic_facet antibody
cell enzyme
DNA
glycosidase inhibitor
hydroxyurea
poly(adenosine diphosphate ribose)
poly(adenosine diphosphate ribose) glycohydrolase
RNA
unclassified drug
article
catalysis
cell cycle progression
cell nucleus
Chagas disease
concentration (parameters)
DNA damage
electron microscopy
enzyme localization
epimastigote
gene
gene expression
genetic analysis
genetic conservation
growth rate
host cell
human
human cell
immunofluorescence test
in vitro study
kinetoplastid life cycle stage
Kinetoplastida
nonhuman
nucleotide sequence
RNA interference
TcPARG gene
Trypanosoma cruzi
trypomastigote
Vero cell
Western blotting
Adenosine Diphosphate
Animals
Blotting, Northern
Blotting, Southern
Blotting, Western
Catalysis
Cell Cycle
Cell Line, Tumor
Cercopithecus aethiops
Chagas Disease
Fluorescent Antibody Technique, Indirect
Glycoside Hydrolases
Humans
Hydroxyurea
Life Cycle Stages
Microscopy, Electron
Pyrrolidines
Trypanosoma cruzi
Vero Cells
Mammalia
Trypanosoma cruzi
Trypanosomatidae
description Trypanosoma cruzi, etiological agent of Chagas' disease, has a complex life cycle which involves the invasion of mammalian host cells, differentiation and intracellular replication. Here we report the first insights into the biological role of a poly(ADP-ribose) glycohydrolase in a trypanosomatid (TcPARG). In silico analysis of the TcPARG gene pointed out the conservation of key residues involved in the catalytic process and, by Western blot, we demonstrated that it is expressed in a life stage-dependant manner. Indirect immunofluorescence assays and electron microscopy using an anti-TcPARG antibody showed that this enzyme is localized in the nucleus independently of the presence of DNA damage or cell cycle stage. The addition of poly(ADP-ribose) glycohydrolase inhibitors ADP-HPD (adenosine diphosphate (hydroxymethyl) pyrrolidinediol) or DEA (6,9-diamino-2-ethoxyacridine lactate monohydrate) to the culture media, both at a 1 μM concentration, reduced in vitro epimastigote growth by 35% and 37% respectively, when compared to control cultures. We also showed that ADP-HPD 1 μM can lead to an alteration in the progression of the cell cycle in hydroxyurea synchronized cultures of T. cruzi epimastigotes. Outstandingly, here we demonstrate that the lack of poly(ADP-ribose) glycohydrolase activity in Vero and A549 host cells, achieved by chemical inhibition or iRNA, produces the reduction of the percentage of infected cells as well as the number of amastigotes per cell and trypomastigotes released, leading to a nearly complete abrogation of the infection process. We conclude that both, T. cruzi and the host, poly(ADP-ribose) glycohydrolase activities are important players in the life cycle of Trypanosoma cruzi, emerging as a promising therapeutic target for the treatment of Chagas' disease. © 2013 Vilchez Larrea et al.
format Artículo
Artículo
publishedVersion
author Vilchez Larrea, S.C.
Schlesinger, M.
Kevorkian, M.L.
Flawiá, M.M.
Alonso, G.D.
Fernández Villamil, S.H.
author_facet Vilchez Larrea, S.C.
Schlesinger, M.
Kevorkian, M.L.
Flawiá, M.M.
Alonso, G.D.
Fernández Villamil, S.H.
author_sort Vilchez Larrea, S.C.
title Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle
title_short Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle
title_full Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle
title_fullStr Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle
title_full_unstemmed Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle
title_sort host cell poly(adp-ribose) glycohydrolase is crucial for trypanosoma cruzi infection cycle
publishDate 2013
url http://hdl.handle.net/20.500.12110/paper_19326203_v8_n6_p_VilchezLarrea
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