Protein engineering of Saccharomyces cerevisiae transporter Pdr5p identifies key residues that impact Fusarium mycotoxin export and resistance to inhibition
Abstract Cereal infection by the broad host range fungal pathogen Fusarium graminearum is a significant global agricultural and food safety issue due to the deposition of mycotoxins within infected grains. Methods to study the intracellular effects of mycotoxins often use the baker's yeast mode...
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doaj-e61293b677054e77b363f3a7c704cbaa2021-09-01T17:00:30ZengWileyMicrobiologyOpen2045-88272016-12-015697999110.1002/mbo3.381Protein engineering of Saccharomyces cerevisiae transporter Pdr5p identifies key residues that impact Fusarium mycotoxin export and resistance to inhibitionAmanda B. Gunter0Anne Hermans1Whynn Bosnich2Douglas A. Johnson3Linda J. Harris4Steve Gleddie5Ottawa Research and Development Centre Agriculture and Agri‐Food Canada Ottawa ON K1A 0C6 CanadaOttawa Research and Development Centre Agriculture and Agri‐Food Canada Ottawa ON K1A 0C6 CanadaOttawa Research and Development Centre Agriculture and Agri‐Food Canada Ottawa ON K1A 0C6 CanadaOttawa‐Carleton Institute of Biology University of Ottawa Ottawa ON K1N 6N5 CanadaOttawa Research and Development Centre Agriculture and Agri‐Food Canada Ottawa ON K1A 0C6 CanadaOttawa Research and Development Centre Agriculture and Agri‐Food Canada Ottawa ON K1A 0C6 CanadaAbstract Cereal infection by the broad host range fungal pathogen Fusarium graminearum is a significant global agricultural and food safety issue due to the deposition of mycotoxins within infected grains. Methods to study the intracellular effects of mycotoxins often use the baker's yeast model system (Saccharomyces cerevisiae); however, this organism has an efficient drug export network known as the pleiotropic drug resistance (PDR) network, which consists of a family of multidrug exporters. This study describes the first study that has evaluated the potential involvement of all known or putative ATP‐binding cassette (ABC) transporters from the PDR network in exporting the F. graminearum trichothecene mycotoxins deoxynivalenol (DON) and 15‐acetyl‐deoxynivalenol (15A‐DON) from living yeast cells. We found that Pdr5p appears to be the only transporter from the PDR network capable of exporting these mycotoxins. We engineered mutants of Pdr5p at two sites previously identified as important in determining substrate specificity and inhibitor susceptibility. These results indicate that it is possible to alter inhibitor insensitivity while maintaining the ability of Pdr5p to export the mycotoxins DON and 15A‐DON, which may enable the development of resistance strategies to generate more Fusarium‐tolerant crop plants.https://doi.org/10.1002/mbo3.381ABC transportersdeoxynivalenoldrug resistanceenniatinFusarium graminearumyeast |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Amanda B. Gunter Anne Hermans Whynn Bosnich Douglas A. Johnson Linda J. Harris Steve Gleddie |
spellingShingle |
Amanda B. Gunter Anne Hermans Whynn Bosnich Douglas A. Johnson Linda J. Harris Steve Gleddie Protein engineering of Saccharomyces cerevisiae transporter Pdr5p identifies key residues that impact Fusarium mycotoxin export and resistance to inhibition MicrobiologyOpen ABC transporters deoxynivalenol drug resistance enniatin Fusarium graminearum yeast |
author_facet |
Amanda B. Gunter Anne Hermans Whynn Bosnich Douglas A. Johnson Linda J. Harris Steve Gleddie |
author_sort |
Amanda B. Gunter |
title |
Protein engineering of Saccharomyces cerevisiae transporter Pdr5p identifies key residues that impact Fusarium mycotoxin export and resistance to inhibition |
title_short |
Protein engineering of Saccharomyces cerevisiae transporter Pdr5p identifies key residues that impact Fusarium mycotoxin export and resistance to inhibition |
title_full |
Protein engineering of Saccharomyces cerevisiae transporter Pdr5p identifies key residues that impact Fusarium mycotoxin export and resistance to inhibition |
title_fullStr |
Protein engineering of Saccharomyces cerevisiae transporter Pdr5p identifies key residues that impact Fusarium mycotoxin export and resistance to inhibition |
title_full_unstemmed |
Protein engineering of Saccharomyces cerevisiae transporter Pdr5p identifies key residues that impact Fusarium mycotoxin export and resistance to inhibition |
title_sort |
protein engineering of saccharomyces cerevisiae transporter pdr5p identifies key residues that impact fusarium mycotoxin export and resistance to inhibition |
publisher |
Wiley |
series |
MicrobiologyOpen |
issn |
2045-8827 |
publishDate |
2016-12-01 |
description |
Abstract Cereal infection by the broad host range fungal pathogen Fusarium graminearum is a significant global agricultural and food safety issue due to the deposition of mycotoxins within infected grains. Methods to study the intracellular effects of mycotoxins often use the baker's yeast model system (Saccharomyces cerevisiae); however, this organism has an efficient drug export network known as the pleiotropic drug resistance (PDR) network, which consists of a family of multidrug exporters. This study describes the first study that has evaluated the potential involvement of all known or putative ATP‐binding cassette (ABC) transporters from the PDR network in exporting the F. graminearum trichothecene mycotoxins deoxynivalenol (DON) and 15‐acetyl‐deoxynivalenol (15A‐DON) from living yeast cells. We found that Pdr5p appears to be the only transporter from the PDR network capable of exporting these mycotoxins. We engineered mutants of Pdr5p at two sites previously identified as important in determining substrate specificity and inhibitor susceptibility. These results indicate that it is possible to alter inhibitor insensitivity while maintaining the ability of Pdr5p to export the mycotoxins DON and 15A‐DON, which may enable the development of resistance strategies to generate more Fusarium‐tolerant crop plants. |
topic |
ABC transporters deoxynivalenol drug resistance enniatin Fusarium graminearum yeast |
url |
https://doi.org/10.1002/mbo3.381 |
work_keys_str_mv |
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