TcPARP: A DNA damage-dependent poly(ADP-ribose) polymerase from Trypanosoma cruzi
Poly(ADP-ribose) polymerase (PARP) is a nuclear enzyme present in most eukaryotes and has been involved in processes such as DNA repair and gene expression. The poly(ADP-ribose) polymer (PAR) is mainly catabolised by poly(ADP-ribose) glycohydrolase. Here, we describe the cloning and characterisation...
Guardado en:
| Publicado: |
2008
|
|---|---|
| Materias: | |
| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00207519_v38_n3-4_p277_FernandezVillamil http://hdl.handle.net/20.500.12110/paper_00207519_v38_n3-4_p277_FernandezVillamil |
| Aporte de: |
| id |
paper:paper_00207519_v38_n3-4_p277_FernandezVillamil |
|---|---|
| record_format |
dspace |
| spelling |
paper:paper_00207519_v38_n3-4_p277_FernandezVillamil2023-06-08T14:41:26Z TcPARP: A DNA damage-dependent poly(ADP-ribose) polymerase from Trypanosoma cruzi DNA repair signalling PARG PARP Trypanosoma cruzi 3 aminobenzamide beta lapachone histone hydrogen peroxide menadione nicotinamide nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase theophylline thymidine clone damage DNA enzyme activity gene expression inhibition protozoan recombination article catalysis DNA damage enzyme activity gene expression in vivo study molecular cloning nonhuman nucleotide sequence protein analysis protein synthesis sequence analysis Trypanosoma cruzi Animals Base Sequence Cloning, Molecular DNA Damage DNA Repair Electrophoresis, Polyacrylamide Gel Enzyme Activation Escherichia coli Gene Expression Immunoblotting Immunohistochemistry Life Cycle Stages Molecular Sequence Data Parasitology Poly Adenosine Diphosphate Ribose Poly(ADP-ribose) Polymerases Sequence Analysis, DNA Trypanosoma cruzi Eukaryota Trypanosoma cruzi Trypanosomatidae Poly(ADP-ribose) polymerase (PARP) is a nuclear enzyme present in most eukaryotes and has been involved in processes such as DNA repair and gene expression. The poly(ADP-ribose) polymer (PAR) is mainly catabolised by poly(ADP-ribose) glycohydrolase. Here, we describe the cloning and characterisation of a PARP from Trypanosoma cruzi (TcPARP). The recombinant enzyme (Mr = 65) required DNA for catalytic activity and it was strongly enhanced by nicked DNA. Histones purified from T. cruzi increased TcPARP activity and the covalent attachment of [32P]ADP-ribose moieties to histones was demonstrated. TcPARP required no magnesium or any other metal ion cofactor for its activity. The enzyme was inhibited by 3-aminobenzamide, nicotinamide, theophylline and thymidine but not by menadione. We demonstrated an automodification reaction of TcPARP, and that the removal of attached PAR from this protein resulted in an increase of its activity. The enzyme was expressed in all parasite stages (amastigotes, epimastigotes and trypomastigotes). When T. cruzi epimastigotes were exposed to DNA-damaging agents such as hydrogen peroxide or β-lapachone, PAR drastically increased in the nucleus, thus confirming PAR synthesis in vivo and suggesting a physiological role for PARP in trypanosomatid DNA repair signalling. © 2007 Australian Society for Parasitology Inc. 2008 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00207519_v38_n3-4_p277_FernandezVillamil http://hdl.handle.net/20.500.12110/paper_00207519_v38_n3-4_p277_FernandezVillamil |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
DNA repair signalling PARG PARP Trypanosoma cruzi 3 aminobenzamide beta lapachone histone hydrogen peroxide menadione nicotinamide nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase theophylline thymidine clone damage DNA enzyme activity gene expression inhibition protozoan recombination article catalysis DNA damage enzyme activity gene expression in vivo study molecular cloning nonhuman nucleotide sequence protein analysis protein synthesis sequence analysis Trypanosoma cruzi Animals Base Sequence Cloning, Molecular DNA Damage DNA Repair Electrophoresis, Polyacrylamide Gel Enzyme Activation Escherichia coli Gene Expression Immunoblotting Immunohistochemistry Life Cycle Stages Molecular Sequence Data Parasitology Poly Adenosine Diphosphate Ribose Poly(ADP-ribose) Polymerases Sequence Analysis, DNA Trypanosoma cruzi Eukaryota Trypanosoma cruzi Trypanosomatidae |
| spellingShingle |
DNA repair signalling PARG PARP Trypanosoma cruzi 3 aminobenzamide beta lapachone histone hydrogen peroxide menadione nicotinamide nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase theophylline thymidine clone damage DNA enzyme activity gene expression inhibition protozoan recombination article catalysis DNA damage enzyme activity gene expression in vivo study molecular cloning nonhuman nucleotide sequence protein analysis protein synthesis sequence analysis Trypanosoma cruzi Animals Base Sequence Cloning, Molecular DNA Damage DNA Repair Electrophoresis, Polyacrylamide Gel Enzyme Activation Escherichia coli Gene Expression Immunoblotting Immunohistochemistry Life Cycle Stages Molecular Sequence Data Parasitology Poly Adenosine Diphosphate Ribose Poly(ADP-ribose) Polymerases Sequence Analysis, DNA Trypanosoma cruzi Eukaryota Trypanosoma cruzi Trypanosomatidae TcPARP: A DNA damage-dependent poly(ADP-ribose) polymerase from Trypanosoma cruzi |
| topic_facet |
DNA repair signalling PARG PARP Trypanosoma cruzi 3 aminobenzamide beta lapachone histone hydrogen peroxide menadione nicotinamide nicotinamide adenine dinucleotide adenosine diphosphate ribosyltransferase theophylline thymidine clone damage DNA enzyme activity gene expression inhibition protozoan recombination article catalysis DNA damage enzyme activity gene expression in vivo study molecular cloning nonhuman nucleotide sequence protein analysis protein synthesis sequence analysis Trypanosoma cruzi Animals Base Sequence Cloning, Molecular DNA Damage DNA Repair Electrophoresis, Polyacrylamide Gel Enzyme Activation Escherichia coli Gene Expression Immunoblotting Immunohistochemistry Life Cycle Stages Molecular Sequence Data Parasitology Poly Adenosine Diphosphate Ribose Poly(ADP-ribose) Polymerases Sequence Analysis, DNA Trypanosoma cruzi Eukaryota Trypanosoma cruzi Trypanosomatidae |
| description |
Poly(ADP-ribose) polymerase (PARP) is a nuclear enzyme present in most eukaryotes and has been involved in processes such as DNA repair and gene expression. The poly(ADP-ribose) polymer (PAR) is mainly catabolised by poly(ADP-ribose) glycohydrolase. Here, we describe the cloning and characterisation of a PARP from Trypanosoma cruzi (TcPARP). The recombinant enzyme (Mr = 65) required DNA for catalytic activity and it was strongly enhanced by nicked DNA. Histones purified from T. cruzi increased TcPARP activity and the covalent attachment of [32P]ADP-ribose moieties to histones was demonstrated. TcPARP required no magnesium or any other metal ion cofactor for its activity. The enzyme was inhibited by 3-aminobenzamide, nicotinamide, theophylline and thymidine but not by menadione. We demonstrated an automodification reaction of TcPARP, and that the removal of attached PAR from this protein resulted in an increase of its activity. The enzyme was expressed in all parasite stages (amastigotes, epimastigotes and trypomastigotes). When T. cruzi epimastigotes were exposed to DNA-damaging agents such as hydrogen peroxide or β-lapachone, PAR drastically increased in the nucleus, thus confirming PAR synthesis in vivo and suggesting a physiological role for PARP in trypanosomatid DNA repair signalling. © 2007 Australian Society for Parasitology Inc. |
| title |
TcPARP: A DNA damage-dependent poly(ADP-ribose) polymerase from Trypanosoma cruzi |
| title_short |
TcPARP: A DNA damage-dependent poly(ADP-ribose) polymerase from Trypanosoma cruzi |
| title_full |
TcPARP: A DNA damage-dependent poly(ADP-ribose) polymerase from Trypanosoma cruzi |
| title_fullStr |
TcPARP: A DNA damage-dependent poly(ADP-ribose) polymerase from Trypanosoma cruzi |
| title_full_unstemmed |
TcPARP: A DNA damage-dependent poly(ADP-ribose) polymerase from Trypanosoma cruzi |
| title_sort |
tcparp: a dna damage-dependent poly(adp-ribose) polymerase from trypanosoma cruzi |
| publishDate |
2008 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00207519_v38_n3-4_p277_FernandezVillamil http://hdl.handle.net/20.500.12110/paper_00207519_v38_n3-4_p277_FernandezVillamil |
| _version_ |
1768543452876767232 |