Haemonchus contortus:Cloning and Functional Expression of a cDNA Encoding Ornithine Decarboxylase and Development of a Screen for Inhibitors

Klein, Ronald D.; Favreau, Mitchell; Alexander-Bowman, Susan J.; Nulf, Susan C.; Vanover, L.; Winterrowd, C.A.; Yarlett, Nigel; Martinez, Martha P.; Keithly, Janet S.; Zantello, Marjorie R.; Thomas, Ernst; Geary, Timothy G.

doi:10.1006/expr.1997.4213

Cited by 17 publications

(18 citation statements)

References 46 publications

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“…For instance, Geary and co-workers expressed potential drug targets for Haemonchus contortus (wireworm, an important parasite of ruminants) in Escherichia coli [36], [37] and later in Saccharomyces cerevisiae [38], and pioneered the development of high-throughput drug screens for antiparasitics in yeast [38].…”

Section: Introductionmentioning

confidence: 99%

Functional Expression of Parasite Drug Targets and Their Human Orthologs in Yeast

et al. 2011

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BackgroundThe exacting nutritional requirements and complicated life cycles of parasites mean that they are not always amenable to high-throughput drug screening using automated procedures. Therefore, we have engineered the yeast Saccharomyces cerevisiae to act as a surrogate for expressing anti-parasitic targets from a range of biomedically important pathogens, to facilitate the rapid identification of new therapeutic agents.Methodology/Principal FindingsUsing pyrimethamine/dihydrofolate reductase (DHFR) as a model parasite drug/drug target system, we explore the potential of engineered yeast strains (expressing DHFR enzymes from Plasmodium falciparum, P. vivax, Homo sapiens, Schistosoma mansoni, Leishmania major, Trypanosoma brucei and T. cruzi) to exhibit appropriate differential sensitivity to pyrimethamine. Here, we demonstrate that yeast strains (lacking the major drug efflux pump, Pdr5p) expressing yeast (ScDFR1), human (HsDHFR), Schistosoma (SmDHFR), and Trypanosoma (TbDHFR and TcDHFR) DHFRs are insensitive to pyrimethamine treatment, whereas yeast strains producing Plasmodium (PfDHFR and PvDHFR) DHFRs are hypersensitive. Reassuringly, yeast strains expressing field-verified, drug-resistant mutants of P. falciparum DHFR (Pfdhfr 51I,59R,108N) are completely insensitive to pyrimethamine, further validating our approach to drug screening. We further show the versatility of the approach by replacing yeast essential genes with other potential drug targets, namely phosphoglycerate kinases (PGKs) and N-myristoyl transferases (NMTs).Conclusions/SignificanceWe have generated a number of yeast strains that can be successfully harnessed for the rapid and selective identification of urgently needed anti-parasitic agents.

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Section: Introductionmentioning

confidence: 99%

Functional Expression of Parasite Drug Targets and Their Human Orthologs in Yeast

et al. 2011

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“…However, the first report of the use of a substitution platform in HTS was against the ornithine decarboxylase (ODC), a key regulatory enzyme in eukaryotic polyamine (PA) biosynthesis, from Haemonchus contortus, a nematode worm that is a major parasite of ruminants. Similar to above, the platform utilized a S. cerevisae ODC deficient mutant complemented by expression of the H. contortus orthologue [35] [36,37,38]. An S. cerevisiae substitution platform has also been used in HTS to identify inhibitors of the anti-leishmanial drug target inositol phosphorylceramide (IPC) synthase [39,40,41].…”

Section: Substitutionmentioning

confidence: 99%

“…The necessity of the secondary assay approach, target directed and across both in vitro and in cellulo platforms, was illustrated very clearly in one of the very first yeast-based HTS campaigns from 1997 [35]. Klein et al identified a single, selective, compound following HTS and counter screening in conditions where the target, helminth ODC, is redundant.…”

Section: Secondary Assaysmentioning

confidence: 99%

Yeast: bridging the gap between phenotypic and biochemical assays for high-throughput screening

Denny

2018

Expert Opinion on Drug Discovery

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“…ODC-deficient yeasts complemented with the nematode-ODC gene were used to test a 90,000-smallmolecule library with the unexpected result of just a single and wrong hit, stilbamidine, which is a well-known inhibitor of S-adenosylmethionine decarboxylase -an important target of the polyamine biosynthesis pathway -but not of ODC. [18] After this disappointing failure, yeast-based platforms were not resumed again to screen parasite targets until very recently. Dihydrofolate reductase (DHFR) is a recognized target that participates in thymidine, histidine, and methionine synthesis in several major parasites.…”

Section: Approaches To Target Identificationmentioning

confidence: 99%

Yeast-based systems for tropical disease drug discovery

Balaña‐Fouce

Reguera

2016

Expert Opinion on Drug Discovery

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Haemonchus contortus:Cloning and Functional Expression of a cDNA Encoding Ornithine Decarboxylase and Development of a Screen for Inhibitors

Cited by 17 publications

References 46 publications

Functional Expression of Parasite Drug Targets and Their Human Orthologs in Yeast

Functional Expression of Parasite Drug Targets and Their Human Orthologs in Yeast

Yeast: bridging the gap between phenotypic and biochemical assays for high-throughput screening

Yeast-based systems for tropical disease drug discovery

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