Systematic phenotyping of a large-scale Candida glabrata deletion collection reveals novel antifungal tolerance genes.

Tobias Ma (1) , Biao Ma (2, 3) , Ekkehard Hiller (4) , Fabian Istel (1) , Michael Tscherner (1) , Sascha Brunke (5, 6, 7) , Lauren Ames (8, 3) , Arnaud Firon (9) , Brian Green (2) , Vitor Cabral (10, 9) , Marina Marcet-Houben (11) , Ilse D Jacobsen (6, 7, 5) , Jessica Quintin (12) , Katja Seider (6) , Ingrid Frohner (1) , Walter Glaser (1) , Helmut Jungwirth (13) , Sophie Bachellier-Bassi (9) , Murielle Chauvel (9) , Ute Zeidler (9) , Dominique Ferrandon (12) , Toni Gabaldón (11) , Bernhard Hube (6, 7, 5) , Christophe d'Enfert (9) , Steffen Rupp (4) , Brendan Cormack (2) , Ken Haynes (3, 8) , Karl Kuchler (1)

1 Medizinische Universität Wien = Medical University of Vienna
2 Johns Hopkins University School of Medicine [Baltimore]
3 Imperial College London
4 Fraunhofer IGB - Fraunhofer Institute for Interfacial Engineering and Biotechnology
5 Center for sepsis control care
6 Leibniz Institute for Natural Product Research and Infection Biology (Hans Knoell Institute)
7 Friedrich-Schiller-Universität = Friedrich Schiller University Jena [Jena, Germany]
8 University of Exeter
9 BPF - Biologie et Pathogénicité fongiques
10 Cellule Pasteur UPMC
11 CRG - Centre for Genomic Regulation [Barcelona]
12 IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire
13 Institut für Molekulare Biowissenschaften [Graz]

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.

Domains: Mycology

Tobias Ma, Biao Ma, Ekkehard Hiller, Fabian Istel, Michael Tscherner, et al.. Systematic phenotyping of a large-scale Candida glabrata deletion collection reveals novel antifungal tolerance genes.. PLoS Pathogens, 2014, 10 (6), pp.e1004211. ⟨10.1371/journal.ppat.1004211⟩. ⟨pasteur-01522550⟩