Detection and characterization of DNA polymerase activity in Entamoeba histolytica.
DNA polymerase activity was detected and characterized in nuclear extracts from trophozoites of Entamoeba histolytica. The activity was high at pH 2 to pH 6, but at pH 8 and 10 the activity was very low. The presence of K+ was inhibitory for the activity and a higher concentration of K+ markedly inhibited the activity. Magnesium ions (Mg2+) were absolutely required for activity and its optimal concentration was 6 to 8 mM. The activity was markedly inhibited by aphidicolin which is an inhibitor of mammalian DNA polymerases alpha, delta, and epsilon and also by N-ethylmaleimide which is an inhibitor of DNA polymerases, alpha, gamma, delta and epsilon. However, inhibition of the activity by 2', 3'-dideoxythymidine-5'-triphosphate which is an inhibitor of DNA polymerases beta and gamma was relatively weak. Thus sensitivity of the E. histolytica enzyme to these inhibitors was similar to that of mammalian DNA polymerases (alpha, delta and epsilon) of the alpha family. Monoclonal antibodies against human DNA polymerase alpha did not bind to DNA polymerase of E. histolytica.
Makioka
A
A
Department of Tropical Medicine, Jikei University School of Medicine, Tokyo, Japan.
Kobayashi
S
S
Takeuchi
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000913:Antibodies, Protozoan / Q000276:immunology; D016590:Aphidicolin / Q000494:pharmacology; D002457:Cell Extracts; D004259:DNA-Directed DNA Polymerase / Q000276:immunology / Q000378:metabolism*; D054306:Dideoxynucleotides; D004748:Entamoeba histolytica / Q000201:enzymology*; D005033:Ethylmaleimide / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D019384:Nucleic Acid Synthesis Inhibitors; D011188:Potassium / Q000494:pharmacology; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013942:Thymine Nucleotides / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D002457:Cell Extracts; D054306:Dideoxynucleotides; D019384:Nucleic Acid Synthesis Inhibitors; D015800:Protozoan Proteins; D013942:Thymine Nucleotides; D016590:Aphidicolin; D004259:DNA-Directed DNA Polymerase; D008274:Magnesium; C017216:2',3'-dideoxythymidine triphosphate; D005033:Ethylmaleimide; D011188:Potassium