Isolation and partial characterization of three DNA polymerases from Trypanosoma cruzi.
Three distinct DNA polymerase fractions (A, B and C), were isolated from Trypanosoma cruzi epimastigote forms. Fraction A is a low molecular mass enzyme corresponding to beta-like DNA polymerase of T. cruzi. Fraction B co-purified along several purification steps with fraction A, but in the last step it was clearly separated by a phosphocellulose chromatography. Fraction C was separated from fractions A and B by binding to DEAE-cellulose column, since the other two fractions were eluted in the flowthrough. This enzyme has an apparent native molecular mass of 100 kDa and showed a high preference for poly(dC)-oligo(dG) among different template-primers tested as substrate. Western-blot and biochemical analysis strongly suggest that the three DNA polymerase fractions correspond to different molecular entities. These results are in agreement with the idea that fraction C is a new DNA polymerase of T. cruzi, not described before.
Venegas
J
J
Programa tide Parasitología, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago. jvenega@canela.med.uchile.cl
Salas
J
J
Gonzalez
C
C
Zulantay
I
I
Diaz
E
E
Gajardo
M
M
Sanchez
G
G
Solari
A
A
D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D015153:Blotting, Western; D002499:Centrifugation, Density Gradient; D002848:Chromatography, DEAE-Cellulose; D004258:DNA Polymerase III / Q000737:chemistry* / Q000302:isolation & purification; D019951:DNA Polymerase beta / Q000737:chemistry* / Q000302:isolation & purification; D003848:Deoxyguanine Nucleotides / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D020128:Inhibitory Concentration 50; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D014349:Trypanosoma cruzi / Q000201:enzymology*
D003848:Deoxyguanine Nucleotides; D004791:Enzyme Inhibitors; D016590:Aphidicolin; C040364:N(2)-(4-n-butylphenyl)-2'-deoxyguanosine 5'-triphosphate; C065639:DNA polymerase A; C040220:DNA polymerase C; D009713:Nucleotidyltransferases; D004258:DNA Polymerase III; D019951:DNA Polymerase beta; D005033:Ethylmaleimide