Article Figures & Data
Figures
- FIG. 1.
Amino acid sequence alignment of Fks1p containing “hot-spot 1” regions from five fungal species: Saccharomyces cerevisiae (Sc), Candida glabrata (Cg), Candida albicans (Ca), Aspergillus nidulans (An), and Aspergillus fumigatus (Af). The highly conserved Ser locus is shaded.
- FIG. 2.
CSF-sensitive and -resistant isolates of A. fumigatus. (A) Growth of strains EMFR-S678P (mutant) and KU80Δ (wild type) on AM3 with 10.0 μg/ml of CSF plus pyrimidine. (B) Growth of EMFR-S678P/WT containing plasmid-expressed fks1(S678P) (top set). EMFR-FKS1-Restored isolates 1 to 4 with restored sensitivity to echinocandin following loss of plasmid (bottom set). Strains were grown for 72 h at 37°C on AM3 with 10.0 μg/ml of CSF.
- FIG. 3.
Echinocandin inhibition profiles of enriched GS complexes from the wild type (triangles), EMFR-S678P (circles), and EMFR-FKS1-Restored (squares). The enzymes were evaluated with CSF (A), ANF (B), and MCF (C). The mixed inhibition profile to CSF for GS from EMFR-S678P/WT is also shown (D). Relative GS activities shown in all panels were assessed by the incorporation of [3H]glucose into the radiolabeled product.
Tables
- TABLE 1.
Strains and plasmids used in this study
Strain or plasmid Description Reference(s) or source Strains KU80Δ A. fumigatus pyrGAF::ΔKU80; wild-type strain sensitive to echinocandin drugs 4 EMFR-S678P Homologous recombinant fks1(S678F) mutant of KU80 formed following transformation with linear pRG3-(pyr4)-AMA1-Fks1S678P cut with HpaI This study EMFR-S678P/WT fks1 mutant of KU80 transformed with uncut autonomously replicating plasmid pRG3-(pyr4)-AMA1-Fks1S678P; also contains wild-type FKS1 This study EMFR-FKS1-Restored Strain derived from EMFR-S678P/WT after plasmid eviction This study Plasmids pRG3-AMA1 Contains A. nidulans AMA1 and pyr4 genes 1, 2, 16 pRG3-AMA1-Fks1S678P Contains A. fumigatus fks1(S678P) in pRG3-(pyr4)-AMA1 This study
