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Detection and characterization of DNA polymerase activity in Toxoplasma gondii

Published online by Cambridge University Press:  06 April 2009

A. Makioka ,
B. Stavros ,
J. T. Ellis  and
A. M. Johnson
A. Makioka
Affiliation:
Department of Microbiology, School of Biological and Biomedical Sciences, University of Technology, Sydney, Gore Hill, New South Wales 2065, Australia
B. Stavros
Affiliation:
Department of Microbiology, School of Biological and Biomedical Sciences, University of Technology, Sydney, Gore Hill, New South Wales 2065, Australia
J. T. Ellis
Affiliation:
Department of Microbiology, School of Biological and Biomedical Sciences, University of Technology, Sydney, Gore Hill, New South Wales 2065, Australia
A. M. Johnson
Affiliation:
Department of Microbiology, School of Biological and Biomedical Sciences, University of Technology, Sydney, Gore Hill, New South Wales 2065, Australia
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Summary

A DNA polymerase activity has been detected and characterized in crude extracts from tachzoites of Toxoplasma gondii. The enzyme has a sedimentation coefficient of 6·4 S, corresponding to an approximate molecular weight of 150000 assuming a globular shape. Like mammalian DNA polymerase α, the DNA polymerase of T. gondii was sensitive to N-ethylmaleimide and inhibited by high ionic strength. However, the enzyme activity was not inhibited by aphidicolin which is an inhibitor of mammalian DNA polymerases α, δ and ε and also cytosine-β-D-arabinofuranoside-5′-triphosphate which is an inhibitor of α polymerase. The activity was inhibited by 2′,3′-dideoxythymidine-5′-triphosphate which is an inhibitor of mammalian DNA polymerase β and γ. Magnesium ions (Mg2+) were absolutely required for activity and its optimal concentration was 6 mM. The optimum potassium (K+) concentration was 50 mM and a higher concentration of K+ markedly inhibited the activity. Activity was optimal at pH 8. Monoclonal antibodies against human DNA polymerase did not bind to DNA polymerase of T. gondii. Thus the T. gondii enzyme differs from the human enzymes and may be a useful target for the design of toxoplasmacidal drugs.

Keywords

DNA polymeraseToxoplasma gondiiparasitic protozoa
Type
Research Article
Information
Parasitology , Volume 107 , Issue 2 , August 1993 , pp. 135 - 139
Copyright
Copyright © Cambridge University Press 1993

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