A versatile overexpression strategy in the pathogenic yeast Candida albicans: identification of regulators of morphogenesis and fitness.
Candida albicans is the most frequently encountered human fungal pathogen, causing both superficial infections and life-threatening systemic diseases. Functional genomic studies performed in this organism have mainly used knock-out mutants and extensive collections of overexpression mutants are still lacking. Here, we report the development of a first generation C. albicans ORFeome, the improvement of overexpression systems and the construction of two new libraries of C. albicans strains overexpressing genes for components of signaling networks, in particular protein kinases, protein phosphatases and transcription factors. As a proof of concept, we screened these collections for genes whose overexpression impacts morphogenesis or growth rates in C. albicans. Our screens identified genes previously described for their role in these biological processes, demonstrating the functionality of our strategy, as well as genes that have not been previously associated to these processes. This article emphasizes the potential of systematic overexpression strategies to improve our knowledge of regulatory networks in C. albicans. The C. albicans plasmid and strain collections described here are available at the Fungal Genetics Stock Center. Their extension to a genome-wide scale will represent important resources for the C. albicans community.
Institut Pasteur, Unité Biologie et Pathogénicité Fongiques, Département Génomes et Génétique, Paris, France.
Murielle
M
Chauvel
Audrey
A
Nesseir
Vitor
V
Cabral
Sadri
S
Znaidi
Sophie
S
Goyard
Sophie
S
Bachellier-Bassi
Arnaud
A
Firon
Mélanie
M
Legrand
Dorothée
D
Diogo
Claire
C
Naulleau
Tristan
T
Rossignol
Christophe
C
d'Enfert
D002176:Candida albicans / Q000378:metabolism*; D004318:Doxycycline / Q000494:pharmacology; D005656:Fungal Proteins / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D005800:Genes, Fungal; D016681:Genome, Fungal; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D007700:Kinetics; D008957:Models, Genetic; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic; D015398:Signal Transduction; D013752:Tetracycline / Q000494:pharmacology
D005656:Fungal Proteins; D049452:Green Fluorescent Proteins; D013752:Tetracycline; D004318:Doxycycline