Functional expression of parasite drug targets and their human orthologs in yeast.
The 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 tar...
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ftdoajarticles:oai:doaj.org/article:31e26412041d4689aad0761444da6549 2023-05-15T15:09:28+02:00 Functional expression of parasite drug targets and their human orthologs in yeast. Elizabeth Bilsland Pınar Pir Alex Gutteridge Alexander Johns Ross D King Stephen G Oliver 2011-10-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0001320 https://doaj.org/article/31e26412041d4689aad0761444da6549 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3186757?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001320 https://doaj.org/article/31e26412041d4689aad0761444da6549 PLoS Neglected Tropical Diseases, Vol 5, Iss 10, p e1320 (2011) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2011 ftdoajarticles https://doi.org/10.1371/journal.pntd.0001320 2022-12-31T15:58:43Z The 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.Using 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 ((Sc)DFR1), human ((Hs)DHFR), Schistosoma ((Sm)DHFR), and Trypanosoma ((Tb)DHFR and (Tc)DHFR) DHFRs are insensitive to pyrimethamine treatment, whereas yeast strains producing Plasmodium ((Pf)DHFR and (Pv)DHFR) DHFRs are hypersensitive. Reassuringly, yeast strains expressing field-verified, drug-resistant mutants of P. falciparum DHFR ((Pf)dhfr(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).We have generated a number of yeast strains that can be successfully harnessed for the rapid and selective identification of urgently needed anti-parasitic agents. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 5 10 e1320 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Elizabeth Bilsland Pınar Pir Alex Gutteridge Alexander Johns Ross D King Stephen G Oliver Functional expression of parasite drug targets and their human orthologs in yeast. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
The 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.Using 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 ((Sc)DFR1), human ((Hs)DHFR), Schistosoma ((Sm)DHFR), and Trypanosoma ((Tb)DHFR and (Tc)DHFR) DHFRs are insensitive to pyrimethamine treatment, whereas yeast strains producing Plasmodium ((Pf)DHFR and (Pv)DHFR) DHFRs are hypersensitive. Reassuringly, yeast strains expressing field-verified, drug-resistant mutants of P. falciparum DHFR ((Pf)dhfr(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).We have generated a number of yeast strains that can be successfully harnessed for the rapid and selective identification of urgently needed anti-parasitic agents. |
format |
Article in Journal/Newspaper |
author |
Elizabeth Bilsland Pınar Pir Alex Gutteridge Alexander Johns Ross D King Stephen G Oliver |
author_facet |
Elizabeth Bilsland Pınar Pir Alex Gutteridge Alexander Johns Ross D King Stephen G Oliver |
author_sort |
Elizabeth Bilsland |
title |
Functional expression of parasite drug targets and their human orthologs in yeast. |
title_short |
Functional expression of parasite drug targets and their human orthologs in yeast. |
title_full |
Functional expression of parasite drug targets and their human orthologs in yeast. |
title_fullStr |
Functional expression of parasite drug targets and their human orthologs in yeast. |
title_full_unstemmed |
Functional expression of parasite drug targets and their human orthologs in yeast. |
title_sort |
functional expression of parasite drug targets and their human orthologs in yeast. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2011 |
url |
https://doi.org/10.1371/journal.pntd.0001320 https://doaj.org/article/31e26412041d4689aad0761444da6549 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 5, Iss 10, p e1320 (2011) |
op_relation |
http://europepmc.org/articles/PMC3186757?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0001320 https://doaj.org/article/31e26412041d4689aad0761444da6549 |
op_doi |
https://doi.org/10.1371/journal.pntd.0001320 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
5 |
container_issue |
10 |
container_start_page |
e1320 |
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1766340655006613504 |