Trypanosoma cruzi nucleoside diphosphate kinase 1 ( TcNDPK1) has a broad nuclease activity.
Here, we present the characterization of a trypanosomatid nucleoside diphosphate kinase (TcNDPK1) exhibiting nuclease activity. This is the first identification of a NDPK with this property in trypanosomatid organisms. The recombinant TcNDPK1 protein cleaves not only linear DNA, but also supercoiled plasmid DNA. Additionally, TcNDPK1 is capable of degrading Trypanosoma cruzi genomic DNA. ATP or ADP did not affect the nuclease activity, while the absence of Mg2+ completely inhibits this activity. NDPK and nuclease activities were inhibited at the same temperature, suggesting the presence of related catalytic sites. Furthermore, phenogram analysis showed that TcNDPK1 is close to Drosophila melanogaster and human NDPKs. The unspecific nuclease activity could suggest a participation in cellular processes such as programmed cell death.
Miranda
M R
MR
Laboratorio de Biología Molecular de Trypanosoma cruzi (LBMTC), Instituto de Investigaciones Médicas A. Lanari, Universidad de Buenos Aires and CONICET, Combatientes de Malvinas 3150, (1427) Buenos Aires, Argentina.
Canepa
G E
GE
Bouvier
L A
LA
Pereira
C A
CA
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D017124:Conserved Sequence; D016054:DNA, Protozoan / Q000378:metabolism; D004278:DNA, Superhelical / Q000378:metabolism; D006358:Hot Temperature; D008274:Magnesium / Q000378:metabolism; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013997:Time Factors; D014349:Trypanosoma cruzi / Q000145:classification / Q000201:enzymology* / Q000235:genetics
D016054:DNA, Protozoan; D004278:DNA, Superhelical; D011994:Recombinant Proteins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D009701:Nucleoside-Diphosphate Kinase; D008274:Magnesium