Generating genomic platforms to study Candida albicans pathogenesis

Mélanie Legrand; Sophie Bachellier-Bassi; Keunsook K. Lee; Yogesh Chaudhari; Hélène Tournu; Laurence Arbogast; Hélène Boyer; Murielle Chauvel; Vitor Cabral; Corinne Maufrais; Audrey Nesseir; Irena Maslanka; Emmanuelle Permal; Tristan Rossignol; Louise A. Walker; Ute Zeidler; Sadri Znaidi; Floris Schoeters; Charlotte Majgier; Renaud A. Julien; Laurence Ma; Magali Tichit; Christiane Bouchier; Patrick Van Dijck; Carol A. Munro; Christophe d’Enfert

albicans ORFeome project is currently completing its second and third phases, which consist of transferring the 5,102 cloned ORFs into barcoded destination vectors and generating a collection of C. albicans barcoded overexpression mutants, each mutant carrying one of the 5,102 cloned ORFs under the control of the inducible P TET promoter. CandidaOrfDB already provides information on the available overexpression plasmids and strains. In this study, we have paved the way for a second application of the C. albicans ORFeome through the development of tools for a 2H matrix approach of protein-protein interaction detection via mating in C. albicans. Our data show that mating of diploid C. albicans expressing a bait fused to the LexA DNA binding domain and a prey fused to the VP16 activation domain, respectively, allows protein-protein interactions to be tested. In S. cerevisiae, large-scale 2H screens can be performed in a matrix design whereby haploid strains expressing baits and preys are mated and protein-protein interactions are scored in the resulting diploids. Our toolkit now enables the implementation of such a matrix design for large-scale 2H screens in C. albicans. To this aim, a collection of 1500 prey clones has already been generated. As mentioned above, C. albicans codon usage is unusual, which limits the development of applications of the C. albicans ORFeome in species that use standard decoding such as S.