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...

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Detalles Bibliográficos
Autores principales: Fernández Villamil, S.H., Baltanás, R., Alonso, G.D., Vilchez Larrea, S.C., Torres, H.N., Flawiá, M.M.
Formato: JOUR
Materias:
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
in vivo study
molecular cloning
nonhuman
nucleotide sequence
protein analysis
protein synthesis
sequence analysis
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
Eukaryota
Trypanosomatidae
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_00207519_v38_n3-4_p277_FernandezVillamil
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
Descripción
Sumario: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.

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