Doss, Ellen M.Moore, Joshua M.Harman, Bryce H.Doud, Emma H.Rubenstein, Eric M.Bernstein, Douglas A.2024-03-012024-03-012023-08-25Doss EM, Moore JM, Harman BH, Doud EH, Rubenstein EM, Bernstein DA. Characterization of endoplasmic reticulum-associated degradation in the human fungal pathogen Candida albicans. PeerJ. 2023;11:e15897. Published 2023 Aug 25. doi:10.7717/peerj.15897https://hdl.handle.net/1805/38983Background: Candida albicans is the most prevalent human fungal pathogen. In immunocompromised individuals, C. albicans can cause serious systemic disease, and patients infected with drug-resistant isolates have few treatment options. The ubiquitin-proteasome system has not been thoroughly characterized in C. albicans. Research from other organisms has shown ubiquitination is important for protein quality control and regulated protein degradation at the endoplasmic reticulum (ER) via ER-associated protein degradation (ERAD). Methods: Here we perform the first characterization, to our knowledge, of ERAD in a human fungal pathogen. We generated functional knockouts of C. albicans genes encoding three proteins predicted to play roles in ERAD, the ubiquitin ligases Hrd1 and Doa10 and the ubiquitin-conjugating enzyme Ubc7. We assessed the fitness of each mutant in the presence of proteotoxic stress, and we used quantitative tandem mass tag mass spectrometry to characterize proteomic alterations in yeast lacking each gene. Results: Consistent with a role in protein quality control, yeast lacking proteins thought to contribute to ERAD displayed hypersensitivity to proteotoxic stress. Furthermore, each mutant displayed distinct proteomic profiles, revealing potential physiological ERAD substrates, co-factors, and compensatory stress response factors. Among candidate ERAD substrates are enzymes contributing to ergosterol synthesis, a known therapeutic vulnerability of C. albicans. Together, our results provide the first description of ERAD function in C. albicans, and, to our knowledge, any pathogenic fungus.en-USAttribution 4.0 InternationalCandida albicansEndoplasmic reticulum-associated degradationUbiquitin ligaseUbiquitin-conjugating enzymeMass spectrometryHrd1Doa10Ubc7Protein quality controlPathogenic fungiArticle