Primary Structure and Expression of the Dihydropteroate Synthetase Gene of Plasmodium Falciparum

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1994 Jul 19

PMID 8041761

Citations 104

Authors

T Triglia

A F Cowman

Affiliations

Soon will be listed here.

Abstract

The enzyme dihydropteroate synthetase (DHPS) from Plasmodium falciparum is involved in the mechanism of action of the sulfone/sulfonamide group of drugs. We describe the cloning and sequencing of the gene encoding the P. falciparum DHPS enzyme and show that it is a bifunctional enzyme that includes dihydro-6-hydroxymethylpterin pyrophosphokinase (PPPK) at the N terminus of DHPS. The gene encodes a putative protein of 83 kDa that contains two domains that are homologous with the DHPS and PPPK enzymes of other organisms. The PPPK-DHPS gene is encoded on chromosome 8 and has two introns. An antibody raised to the PPPK region of the protein was found to recognize a 68-kDa protein that is expressed throughout the asexual life cycle of the parasite. We have determined the sequence of the DHPS portion of the gene from sulfadoxine-sensitive and -resistant P. falciparum clones and identified sequence differences that may have a role in sulfone/sulfonamide resistance.

Citing Articles

Malaria parasite cysteine and aspartic proteases as key drug targets for antimalarial therapy.

Adeoye A, Lobb K J Mol Model. 2025; 31(3):78.

PMID: 39920505 DOI: 10.1007/s00894-025-06303-0.

To quest new targets of parasite and their potential inhibitors to combat antimalarial drug resistance.

Biswas P, Roy R, Ghosh K, Nath D, Samadder A, Nandi S J Parasit Dis. 2024; 48(4):671-722.

PMID: 39493470 PMC: 11527868. DOI: 10.1007/s12639-024-01687-x.

Combating antimicrobial resistance in malaria, HIV and tuberculosis.

Duffey M, Shafer R, Timm J, Burrows J, Fotouhi N, Cockett M Nat Rev Drug Discov. 2024; 23(6):461-479.

PMID: 38750260 DOI: 10.1038/s41573-024-00933-4.

Putative molecular markers of resistance to antimalarial drugs in malaria parasites from Ghana.

Matrevi S, Adams T, Tandoh K, Opoku-Agyeman P, Bruku S, Ennuson N Front Epidemiol. 2024; 4:1279835.

PMID: 38456076 PMC: 10910922. DOI: 10.3389/fepid.2024.1279835.

Crystal structure of Arabidopsis thaliana HPPK/DHPS, a bifunctional enzyme and target of the herbicide asulam.

Vadlamani G, Sukhoverkov K, Haywood J, Breese K, Fisher M, Stubbs K Plant Commun. 2022; 3(4):100322.

PMID: 35605193 PMC: 9284294. DOI: 10.1016/j.xplc.2022.100322.

References

FERONE R . The enzymic synthesis of dihydropteroate and dihydrofolate by Plasmodium berghei. J Protozool. 1973; 20(3):459-64. DOI: 10.1111/j.1550-7408.1973.tb00926.x. View

LeBlanc S, Wilson C . The dihydroorotate dehydrogenase gene homologue of Plasmodium falciparum. Mol Biochem Parasitol. 1993; 60(2):349-51. DOI: 10.1016/0166-6851(93)90148-q. View

Walter R, Konigk E . Biosynthesis of folic acid compounds in plasmodia. Purification and properties of the 7,8-dihydropteroate-synthesizing enzyme from Plasmodium chabaudi. Hoppe Seylers Z Physiol Chem. 1974; 355(4):431-7. DOI: 10.1515/bchm2.1974.355.1.431. View

FERONE R . Folate metabolism in malaria. Bull World Health Organ. 1977; 55(2-3):291-8. PMC: 2366725. View

TRAGER W, Jenson J . Cultivation of malarial parasites. Nature. 1978; 273(5664):621-2. DOI: 10.1038/273621a0. View

Dieckmann A, Jung A . Mechanisms of sulfadoxine resistance in Plasmodium falciparum. Mol Biochem Parasitol. 1986; 19(2):143-7. DOI: 10.1016/0166-6851(86)90119-2. View

Bianco A, Favaloro J, Burkot T, Culvenor J, Crewther P, Brown G . A repetitive antigen of Plasmodium falciparum that is homologous to heat shock protein 70 of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1986; 83(22):8713-7. PMC: 387001. DOI: 10.1073/pnas.83.22.8713. View

Chu G, Vollrath D, Davis R . Separation of large DNA molecules by contour-clamped homogeneous electric fields. Science. 1986; 234(4783):1582-5. DOI: 10.1126/science.3538420. View

Lopez P, Espinosa M, Greenberg B, Lacks S . Sulfonamide resistance in Streptococcus pneumoniae: DNA sequence of the gene encoding dihydropteroate synthase and characterization of the enzyme. J Bacteriol. 1987; 169(9):4320-6. PMC: 213747. DOI: 10.1128/jb.169.9.4320-4326.1987. View

10.

Smith D, Johnson K . Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988; 67(1):31-40. DOI: 10.1016/0378-1119(88)90005-4. View

11.

Cowman A, Morry M, Biggs B, Cross G, Foote S . Amino acid changes linked to pyrimethamine resistance in the dihydrofolate reductase-thymidylate synthase gene of Plasmodium falciparum. Proc Natl Acad Sci U S A. 1988; 85(23):9109-13. PMC: 282673. DOI: 10.1073/pnas.85.23.9109. View

12.

Krungkrai J, Webster H, Yuthavong Y . De novo and salvage biosynthesis of pteroylpentaglutamates in the human malaria parasite, Plasmodium falciparum. Mol Biochem Parasitol. 1989; 32(1):25-37. DOI: 10.1016/0166-6851(89)90126-6. View

13.

Li W, Bzik D, Gu H, Tanaka M, Fox B, Inselburg J . An enlarged largest subunit of Plasmodium falciparum RNA polymerase II defines conserved and variable RNA polymerase domains. Nucleic Acids Res. 1989; 17(23):9621-36. PMC: 335202. DOI: 10.1093/nar/17.23.9621. View

14.

Foote S, Kyle D, Martin R, Oduola A, Forsyth K, Kemp D . Several alleles of the multidrug-resistance gene are closely linked to chloroquine resistance in Plasmodium falciparum. Nature. 1990; 345(6272):255-8. DOI: 10.1038/345255a0. View

15.

Lopez P, Greenberg B, Lacks S . DNA sequence of folate biosynthesis gene sulD, encoding hydroxymethyldihydropterin pyrophosphokinase in Streptococcus pneumoniae, and characterization of the enzyme. J Bacteriol. 1990; 172(9):4766-74. PMC: 213129. DOI: 10.1128/jb.172.9.4766-4774.1990. View

16.

Slock J, Stahly D, Han C, Six E, Crawford I . An apparent Bacillus subtilis folic acid biosynthetic operon containing pab, an amphibolic trpG gene, a third gene required for synthesis of para-aminobenzoic acid, and the dihydropteroate synthase gene. J Bacteriol. 1990; 172(12):7211-26. PMC: 210846. DOI: 10.1128/jb.172.12.7211-7226.1990. View

17.

Zhang Y, Meshnick S . Inhibition of Plasmodium falciparum dihydropteroate synthetase and growth in vitro by sulfa drugs. Antimicrob Agents Chemother. 1991; 35(2):267-71. PMC: 244989. DOI: 10.1128/AAC.35.2.267. View

18.

Brown H, Coppel R . Primary structure of a Plasmodium falciparum rhoptry antigen. Mol Biochem Parasitol. 1991; 49(1):99-110. DOI: 10.1016/0166-6851(91)90133-q. View

19.

Volpe F, Dyer M, Scaife J, Darby G, Stammers D, Delves C . The multifunctional folic acid synthesis fas gene of Pneumocystis carinii appears to encode dihydropteroate synthase and hydroxymethyldihydropterin pyrophosphokinase. Gene. 1992; 112(2):213-8. DOI: 10.1016/0378-1119(92)90378-3. View

20.

Dallas W, Gowen J, Ray P, Cox M, Dev I . Cloning, sequencing, and enhanced expression of the dihydropteroate synthase gene of Escherichia coli MC4100. J Bacteriol. 1992; 174(18):5961-70. PMC: 207134. DOI: 10.1128/jb.174.18.5961-5970.1992. View