Systematic Functional Elucidation of the Transcription Factor Networks in Candidozyma auris

Candidozyma auris (formerly known as Candida auris), first identified in 2009, has emerged as a multidrug-resistant fungal pathogen associated with high mortality rates, yet its pathogenic mechanisms and virulence factors remain largely unclear. Transcription factors (TFs), key regulators of gene expression with high interspecies divergence, can help uncover the unique biology of C. auris. Hence, we aim to elucidate its transcriptional networks through systematic TF characterization. Using DeepTFactor predictions and subsequent DNA-binding domain examination, we identified 166 putative TF genes and constructed signature-tagged gene-deletion strains for over 80% of these genes in the clade I wild-type (B8441) background. We investigated the in vitro phenotypes of these mutants under various stress conditions, including high temperature, antifungal drugs, and cell membrane/wall stress. Furthermore, we assessed their roles in virulence-related traits, such as biofilm formation and secreted aspartyl protease activity. Building on these in vitro findings, we will extend our analysis to in vivo pathogenicity, employing a high-throughput Caenorhabditis elegans infection screening system, followed by a signature-tagged mutagenesis (STM) murine virulence assay. In conclusion, this study aims to advance the understanding of C. auris virulence mechanisms and identify novel antifungal targets by analyzing its transcriptional networks.