title: Systematic phenotyping of a large-scale Candida glabrata deletion collection reveals novel antifungal tolerance genes.
abstract: The opportunistic fungal pathogen Candida glabrata is a frequent cause of candidiasis, causing infections ranging from superficial to life-threatening disseminated disease. The inherent tolerance of C. glabrata to azole drugs makes this pathogen a serious clinical threat. To identify novel genes implicated in antifungal drug tolerance, we have constructed a large-scale C. glabrata deletion library consisting of 619 unique, individually bar-coded mutant strains, each lacking one specific gene, all together representing almost 12% of the genome. Functional analysis of this library in a series of phenotypic and fitness assays identified numerous genes required for growth of C. glabrata under normal or specific stress conditions, as well as a number of novel genes involved in tolerance to clinically important antifungal drugs such as azoles and echinocandins. We identified 38 deletion strains displaying strongly increased susceptibility to caspofungin, 28 of which encoding proteins that have not previously been linked to echinocandin tolerance. Our results demonstrate the potential of the C. glabrata mutant collection as a valuable resource in functional genomics studies of this important fungal pathogen of humans, and to facilitate the identification of putative novel antifungal drug target and virulence genes.
authors.affiliation: Medical University Vienna, Max F. Perutz Laboratories, Department of Medical Biochemistry, Vienna, Austria.
authors.forename: Tobias
authors.initials: T
authors.lastname: Schwarzmüller
authors.affiliation: Department of Microbiology, Imperial College London, London, United Kingdom; Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Biao
authors.initials: B
authors.lastname: Ma
authors.affiliation: Molekulare Biotechnologie MBT Fraunhofer Institut für Grenzflächen- und Bioverfahrenstechnik IGB Fraunhofer, Stuttgart, Germany.
authors.forename: Ekkehard
authors.initials: E
authors.lastname: Hiller
authors.affiliation: Medical University Vienna, Max F. Perutz Laboratories, Department of Medical Biochemistry, Vienna, Austria.
authors.forename: Fabian
authors.initials: F
authors.lastname: Istel
authors.affiliation: Medical University Vienna, Max F. Perutz Laboratories, Department of Medical Biochemistry, Vienna, Austria.
authors.forename: Michael
authors.initials: M
authors.lastname: Tscherner
authors.affiliation: Department Microbial Pathogenicity Mechanisms, Hans-Knoell-Institute, Jena, Germany; Friedrich Schiller University, Jena, Germany; Center for Sepsis Control and Care, CSCC, Jena University Hospital, Jena, Germany.
authors.forename: Sascha
authors.initials: S
authors.lastname: Brunke
authors.affiliation: Department of Microbiology, Imperial College London, London, United Kingdom; Biosciences, College of Life & Environmental Sciences, University of Exeter, Exeter, United Kingdom.
authors.forename: Lauren
authors.initials: L
authors.lastname: Ames
authors.affiliation: Institut Pasteur, Unité Biologie et Pathogénicité Fongiques, Département Génomes et Génétique, Paris, France; INRA, USC2019, Paris, France.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Firon
authors.affiliation: Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Brian
authors.initials: B
authors.lastname: Green
authors.affiliation: Institut Pasteur, Unité Biologie et Pathogénicité Fongiques, Département Génomes et Génétique, Paris, France; INRA, USC2019, Paris, France; Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France.
authors.forename: Vitor
authors.initials: V
authors.lastname: Cabral
authors.affiliation: Bioinformatics and Genomics Programme, Centre for Genomic Regulation (CRG), Barcelona, Spain.
authors.forename: Marina
authors.initials: M
authors.lastname: Marcet-Houben
authors.affiliation: Department Microbial Pathogenicity Mechanisms, Hans-Knoell-Institute, Jena, Germany; Friedrich Schiller University, Jena, Germany; Center for Sepsis Control and Care, CSCC, Jena University Hospital, Jena, Germany.
authors.forename: Ilse D
authors.initials: ID
authors.lastname: Jacobsen
authors.affiliation: UPR 9022 du CNRS, Université de Strasbourg, Equipe Fondation Recherche Médicale, Institut de Biologie Moléculaire et Cellulaire, Strasbourg, France.
authors.forename: Jessica
authors.initials: J
authors.lastname: Quintin
authors.affiliation: Department Microbial Pathogenicity Mechanisms, Hans-Knoell-Institute, Jena, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Seider
authors.affiliation: Medical University Vienna, Max F. Perutz Laboratories, Department of Medical Biochemistry, Vienna, Austria.
authors.forename: Ingrid
authors.initials: I
authors.lastname: Frohner
authors.affiliation: Medical University Vienna, Max F. Perutz Laboratories, Department of Medical Biochemistry, Vienna, Austria.
authors.forename: Walter
authors.initials: W
authors.lastname: Glaser
authors.affiliation: Institut für Molekulare Biowissenschaften, Universität Graz, Graz, Austria.
authors.forename: Helmut
authors.initials: H
authors.lastname: Jungwirth
authors.affiliation: Institut Pasteur, Unité Biologie et Pathogénicité Fongiques, Département Génomes et Génétique, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Bachellier-Bassi
authors.affiliation: Institut Pasteur, Unité Biologie et Pathogénicité Fongiques, Département Génomes et Génétique, Paris, France.
authors.forename: Murielle
authors.initials: M
authors.lastname: Chauvel
authors.affiliation: Institut Pasteur, Unité Biologie et Pathogénicité Fongiques, Département Génomes et Génétique, Paris, France.
authors.forename: Ute
authors.initials: U
authors.lastname: Zeidler
authors.affiliation: UPR 9022 du CNRS, Université de Strasbourg, Equipe Fondation Recherche Médicale, Institut de Biologie Moléculaire et Cellulaire, Strasbourg, France.
authors.forename: Dominique
authors.initials: D
authors.lastname: Ferrandon
authors.affiliation: Bioinformatics and Genomics Programme, Centre for Genomic Regulation (CRG), Barcelona, Spain; Universitat Pompeu Fabra (UPF), Barcelona, Spain.
authors.forename: Toni
authors.initials: T
authors.lastname: Gabaldón
authors.affiliation: Department Microbial Pathogenicity Mechanisms, Hans-Knoell-Institute, Jena, Germany; Friedrich Schiller University, Jena, Germany; Center for Sepsis Control and Care, CSCC, Jena University Hospital, Jena, Germany.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Hube
authors.affiliation: Institut Pasteur, Unité Biologie et Pathogénicité Fongiques, Département Génomes et Génétique, Paris, France; INRA, USC2019, Paris, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: d'Enfert
authors.affiliation: Molekulare Biotechnologie MBT Fraunhofer Institut für Grenzflächen- und Bioverfahrenstechnik IGB Fraunhofer, Stuttgart, Germany.
authors.forename: Steffen
authors.initials: S
authors.lastname: Rupp
authors.affiliation: Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Brendan
authors.initials: B
authors.lastname: Cormack
authors.affiliation: Department of Microbiology, Imperial College London, London, United Kingdom; Biosciences, College of Life & Environmental Sciences, University of Exeter, Exeter, United Kingdom.
authors.forename: Ken
authors.initials: K
authors.lastname: Haynes
authors.affiliation: Medical University Vienna, Max F. Perutz Laboratories, Department of Medical Biochemistry, Vienna, Austria.
authors.forename: Karl
authors.initials: K
authors.lastname: Kuchler
mesh_terms: D000935:Antifungal Agents / Q000494:pharmacology*; D001393:Azoles / Q000494:pharmacology*; D018441:Biofilms / Q000254:growth & development; D041221:Candida glabrata / Q000235:genetics* / Q000254:growth & development; D002177:Candidiasis / Q000188:drug therapy; D000077336:Caspofungin; D002473:Cell Wall / Q000187:drug effects / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics*; D054714:Echinocandins / Q000494:pharmacology*; D005656:Fungal Proteins / Q000235:genetics; D017353:Gene Deletion; D055786:Gene Knockout Techniques; D015723:Gene Library; D055666:Lipopeptides; D008826:Microbial Sensitivity Tests; D009997:Osmotic Pressure; D010641:Phenotype
chemical_list: D000935:Antifungal Agents; D001393:Azoles; D054714:Echinocandins; D005656:Fungal Proteins; D055666:Lipopeptides; D000077336:Caspofungin