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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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 |
work_keys_str_mv |
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