Journal article 1708 views
Heterologous Expression of Mutated Eburicol 14α-Demethylase (CYP51) Proteins of Mycosphaerella graminicola To Assess Effects on Azole Fungicide Sensitivity and Intrinsic Protein Function
Applied and Environmental Microbiology, Volume: 76, Issue: 9, Pages: 2866 - 2872
Swansea University Authors:
Josie Parker, Diane Kelly, Steven Kelly
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DOI (Published version): 10.1128/aem.02158-09
Abstract
<p><span>The recent decrease in the sensitivity of the Western European population of the wheat pathogen </span><em>Mycosphaerella graminicola</em><span> to azole fungicides has been associated with the emergence and subsequent spread of mutations in the </span...
| Published in: | Applied and Environmental Microbiology |
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| ISSN: | 0099-2240 1098-5336 |
| Published: |
American Society for Microbiology
2010
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| Online Access: |
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| URI: | https://cronfa.swan.ac.uk/Record/cronfa6856 |
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| Abstract: |
<p><span>The recent decrease in the sensitivity of the Western European population of the wheat pathogen </span><em>Mycosphaerella graminicola</em><span> to azole fungicides has been associated with the emergence and subsequent spread of mutations in the </span><em>CYP51</em><span>gene, encoding the azole target sterol 14α-demethylase. In this study, we have expressed wild-type and mutated </span><em>M. graminicola</em><span> CYP51 (MgCYP51) variants in a</span><em>Saccharomyces cerevisiae</em><span> mutant carrying a doxycycline-regulatable </span><em>tetO<sub>7</sub></em><span>-</span><em>CYC</em><span>promoter controlling native </span><em>CYP51</em><span> expression. We have shown that the wild-type MgCYP51 protein complements the function of the orthologous protein in </span><em>S. cerevisiae</em><span>. Mutant MgCYP51 proteins containing amino acid alterations L50S, Y459D, and Y461H and the two-amino-acid deletion ΔY459/G460, commonly identified in modern </span><em>M. graminicola</em><span> populations, have no effect on the capacity of the</span><em>M. graminicola</em><span> protein to function in </span><em>S. cerevisiae</em><span>. We have also shown that the azole fungicide sensitivities of transformants expressing MgCYP51 variants with these alterations are substantially reduced. Furthermore, we have demonstrated that the I381V substitution, correlated with the recent decline in the effectiveness of azoles, destroys the capacity of </span><em>MgCYP51</em><span> to complement the </span><em>S. cerevisiae</em><span> mutant when introduced alone. However, when I381V is combined with changes between residues Y459 and Y461, the function of the </span><em>M. graminicola</em><span> protein is partially restored. These findings demonstrate, for the first time for a plant pathogenic fungus, the impacts that naturally occurring CYP51 alterations have on both azole sensitivity and intrinsic protein function. In addition, we also provide functional evidence underlying the order in which CYP51 alterations in the Western European </span><em>M. graminicola</em><span> population emerged.</span></p> |
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| College: |
Faculty of Medicine, Health and Life Sciences |
| Issue: |
9 |
| Start Page: |
2866 |
| End Page: |
2872 |