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Mutation rates in the dihydrofolate reductase gene of Plasmodium falciparum

Published online by Cambridge University Press:  07 August 2001

S. PAGET-MCNICOL  and
A. SAUL
S. PAGET-MCNICOL
Affiliation:
The University of Queensland and the Malaria and Arbovirus Unit, Queensland Institute of Medical Research, QLD 4029, Australia
A. SAUL
Affiliation:
Malaria Vaccine Development Unit, NIAID/LPD, National Institutes of Health, Rockville, MD 20852, USA
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Abstract

A new method has been established to define the limits on a spontaneous mutation rate for a gene in Plasmodium falciparum. The method combines mathematical modelling and large-scale in vitro culturing and calculates the difference in mutant frequencies at 2 separate time-points. We measured the mutation rate at 2 positions in the dihydrofolate reductase (DHFR) gene of 3D7, a pyrimethamine-sensitive line of P. falciparum. This line was re-cloned and an effectively large population was treated with a selective pyrimethamine concentration of 40 nM. We detected point mutations at codon-46 (TTA to TCA) and codon-108 (AGC to AAC), resulting in serine replacing leucine and asparagine replacing serine respectively in the corresponding gene product. The substitutions caused a decrease in pyrimethamine sensitivity. By mathematical modelling we determined that the mutation rate at a given position in DHFR was low and occurred at less than 2.5×10−9 mutations/DHFR gene/replication. This result has important implications for Plasmodium genetic diversity and anti-malarial drug therapy by demonstrating that even with low mutation rates anti-malarial resistance will inevitably arise when mutant alleles are selected under drug pressure.

Keywords

dihydrofolate reductasemutation ratemalariamathematical modellingPlasmodium falciparum
Type
Research Article
Information
Parasitology , Volume 122 , Issue 5 , May 2001 , pp. 497 - 505
Copyright
2001 Cambridge University Press

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