title: Modeling the Qo site of crop pathogens in Saccharomyces cerevisiae cytochrome b.
abstract: Saccharomyces cerevisiae has been used as a model system to characterize the effect of cytochrome b mutations found in fungal and oomycete plant pathogens resistant to Q(o) inhibitors (QoIs), including the strobilurins, now widely employed in agriculture to control such diseases. Specific residues in the Q(o) site of yeast cytochrome b were modified to obtain four new forms mimicking the Q(o) binding site of Erysiphe graminis, Venturia inaequalis, Sphaerotheca fuliginea and Phytophthora megasperma. These modified versions of cytochrome b were then used to study the impact of the introduction of the G143A mutation on bc(1) complex activity. In addition, the effects of two other mutations F129L and L275F, which also confer levels of QoI insensitivity, were also studied. The G143A mutation caused a high level of resistance to QoI compounds such as myxothiazol, axoxystrobin and pyraclostrobin, but not to stigmatellin. The pattern of resistance conferred by F129L and L275F was different. Interestingly G143A had a slightly deleterious effect on the bc(1) function in V. inaequalis, S. fuliginea and P. megasperma Q(o) site mimics but not in that for E. graminis. Thus small variations in the Q(o) site seem to affect the impact of the G143A mutation on bc(1) activity. Based on this observation in the yeast model, it might be anticipated that the G143A mutation might affect the fitness of pathogens differentially. If so, this could contribute to observed differences in the rates of evolution of QoI resistance in fungal and oomycete pathogens.
authors.affiliation: The Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Fisher
authors.forename: Amanda C
authors.initials: AC
authors.lastname: Brown
authors.forename: Graham
authors.initials: G
authors.lastname: Sexton
authors.forename: Alison
authors.initials: A
authors.lastname: Cook
authors.forename: John
authors.initials: J
authors.lastname: Windass
authors.forename: Brigitte
authors.initials: B
authors.lastname: Meunier
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry / Q000494:pharmacology; D001665:Binding Sites; D002455:Cell Division; D018556:Crops, Agricultural / Q000382:microbiology; D045303:Cytochromes b / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D025141:Drug Resistance, Fungal; D014450:Electron Transport Complex III / Q000378:metabolism; D006873:Hydroquinones / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology* / Q000472:pathogenicity; D017386:Sequence Homology, Amino Acid
chemical_list: D000935:Antifungal Agents; D006873:Hydroquinones; D045303:Cytochromes b; D014450:Electron Transport Complex III