Fig. 4: Mutations in OCA6 and ROB1 enable C. albicans filamentation despite exposure to 1-ABC.

a A selection-based strategy resulted in the identification of six independent lineages with a restored ability of C. albicans to filament in the presence of 1-ABC. The filamentation phenotype was validated in YPD medium at 42 °C in the presence of 1-ABC (300 µM). Data represent two biological replicates. b Summary of whole-genome sequencing results for two of the evolved lineages. All identified loss-of-heterozygosity events or single nucleotide polymorphisms that occurred within open reading frames of each lineage are shown. Results were independently validated using Sanger sequencing. Whole-genome sequencing analysis of Lineages 1 and 2 did not detect any copy number variants relative to the parental strain. Copy number variant analysis was conducted and visualized using the Y-MAP software⁶³. c The OCA6 mutation in Lineage 1 is necessary and sufficient for the filamentation phenotype. A wild-type OCA6 allele from the parental strain (red) was used to replace the mutant OCA6 allele (blue) from Lineage 1. Similarly, the mutant OCA6 allele from Lineage 1 (blue) was used to replace the wild-type OCA6 allele (red) in the parental background. Strains were grown at 42 °C for 6 h and data represent two biological replicates. d The ROB1 mutation in Lineage 2 is necessary and sufficient for the filamentation phenotype. A wild-type ROB1 allele from the parental strain (red) was used to replace the mutant ROB1 allele (blue) from Lineage 2. Similarly, the mutant ROB1 allele from Lineage 2 (blue) was used to replace the wild-type ROB1 allele (red) in the parental background. Strains were grown at 42 °C for 6 h and data represent two biological replicates.