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New Fks hot spot for acquired echinocandin resistance in Saccharomyces cerevisiae and its contribution to intrinsic resistance of Scedosporium species
Journal article   Open access   Peer reviewed

New Fks hot spot for acquired echinocandin resistance in Saccharomyces cerevisiae and its contribution to intrinsic resistance of Scedosporium species

Michael E Johnson, Santosh K Katiyar and Thomas D Edlind
Antimicrobial agents and chemotherapy, v 55(8), pp 3774-3781
Aug 2011
PMID: 21576441
url
https://doi.org/10.1128/AAC.01811-10View
Published, Version of Record (VoR) Open

Abstract

Lipopeptides - pharmacology Saccharomyces cerevisiae - genetics Scedosporium - drug effects Molecular Sequence Data Saccharomyces cerevisiae - drug effects Microbial Sensitivity Tests Cell Wall - drug effects Echinocandins - genetics Base Sequence Caspofungin Echinocandins - pharmacology Glucosyltransferases - genetics Antifungal Agents - pharmacology Glucosyltransferases - antagonists & inhibitors Anidulafungin Membrane Proteins - genetics Cell Wall - chemistry Drug Resistance, Fungal - genetics Saccharomyces cerevisiae Proteins - genetics Micafungin Sequence Analysis, DNA beta-Glucans - metabolism Scedosporium - genetics Sequence Alignment Mutation Candida glabrata - genetics Candida glabrata - drug effects
Echinocandins represent a new antifungal group with potent activity against Candida species. These lipopeptides inhibit the synthesis of β-1,3-glucan, the major cell wall polysaccharide. Acquired resistance or reduced echinocandin susceptibility (RES) is rare and associated with mutations in two "hot spot" regions of Fks1 or Fks2, the probable β-1,3-glucan synthases. In contrast, many fungi demonstrate intrinsic RES for reasons that remain unclear. We are using Saccharomyces cerevisiae to understand the basis for RES by modeling echinocandin-Fks interaction. Previously characterized mutations confer cross-RES; we screened for mutations conferring differential RES, implying direct interaction of that Fks residue with a variable echinocandin side chain. One mutant (in an fks1Δ background) exhibited ≥16-fold micafungin and anidulafungin versus caspofungin RES. Sequencing identified a novel Fks2 mutation, W714L/Y715N. Equivalent W695L/Y696N and related W695L/F/C mutations in Fks1 generated by site-directed mutagenesis and the isolation of a W695L-equivalent mutation in Candida glabrata confirmed the role of the new "hot spot 3" in RES. Further mutagenesis expanded hot spot 3 to Fks1 residues 690 to 700, yielding phenotypes ranging from cross-RES to differential hypersusceptibility. Fks1 sequences from intrinsically RES Scedosporium species revealed W695F-equivalent substitutions; Fks1 hybrids expressing Scedosporium prolificans hot spot 3 confirmed that this substitution imparts RES.

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Web of Science research areas
Microbiology
Pharmacology & Pharmacy
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