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:	Flawiá, Mirtha María, Alonso, Guillermo Daniel
Publicado:	2013
Materias:	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 Vero Cells Mammalia Trypanosomatidae
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19326203_v8_n6_p_VilchezLarrea http://hdl.handle.net/20.500.12110/paper_19326203_v8_n6_p_VilchezLarrea
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_19326203_v8_n6_p_VilchezLarrea
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spelling	paper:paper_19326203_v8_n6_p_VilchezLarrea2023-06-08T16:31:16Z Host Cell Poly(ADP-Ribose) Glycohydrolase Is Crucial for Trypanosoma cruzi Infection Cycle Flawiá, Mirtha María Alonso, Guillermo Daniel 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 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19326203_v8_n6_p_VilchezLarrea 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)
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 Flawiá, Mirtha María Alonso, Guillermo Daniel 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.
author	Flawiá, Mirtha María Alonso, Guillermo Daniel
author_facet	Flawiá, Mirtha María Alonso, Guillermo Daniel
author_sort	Flawiá, Mirtha María
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	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19326203_v8_n6_p_VilchezLarrea http://hdl.handle.net/20.500.12110/paper_19326203_v8_n6_p_VilchezLarrea
work_keys_str_mv	AT flawiamirthamaria hostcellpolyadpriboseglycohydrolaseiscrucialfortrypanosomacruziinfectioncycle AT alonsoguillermodaniel hostcellpolyadpriboseglycohydrolaseiscrucialfortrypanosomacruziinfectioncycle
_version_	1768544706887680000

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

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