Purification and characterization of DNA polymerases from Plasmodium berghei.
DNA polymerases from the malaria parasite Plasmodium berghei were purified more than 50-fold. Several distinct enzymatic activities were isolated that could be distinguished by the use of various specific DNA polymerase inhibitors. In particular, subdivision into an aphidicolin-sensitive and an aphidicolin-resistant group was possible. Further analysis allowed a better comparison with host DNA polymerases and indicated that one aphidicolin-sensitive DNA polymerase resembled DNA polymerase alpha displaying processive DNA synthesis and using RNA primers, whereas another aphidicolin-sensitive DNA polymerase was distributive and only used DNA primers. Marked differences from the host enzymes do exist, however, such as insensitivity to BuPdGTP. Another P. berghei DNA polymerase was isolated that showed characteristics of a DNA polymerase beta-like enzyme, but which differed from host DNA polymerase beta in its insensitivity to dideoxynucleotides.
de Vries
E
E
Department of Tropical Veterinary Medicine and Protozoology, University of Utrecht, The Netherlands.
Stam
J G
JG
Franssen
F F
FF
van der Vliet
P C
PC
Overdulve
J P
JP
D000818:Animals; D016590:Aphidicolin; D002852:Chromatography, Ion Exchange; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D003848:Deoxyguanine Nucleotides / Q000494:pharmacology; D015224:Dideoxynucleosides / Q000494:pharmacology; D004224:Diterpenes / Q000494:pharmacology; D008288:Malaria; D010962:Plasmodium berghei / Q000201:enzymology*
D003848:Deoxyguanine Nucleotides; D015224:Dideoxynucleosides; D004224:Diterpenes; D016590:Aphidicolin; C040364:N(2)-(4-n-butylphenyl)-2'-deoxyguanosine 5'-triphosphate; D004259:DNA-Directed DNA Polymerase