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An Ex vivo Assay to Study Candida albicans Hyphal Morphogenesis in the Gastrointestinal Tract
Published on: July 1, 2020
Proper Sterol Distribution Is Required for Candida albicans Hyphal Formation and Virulence
Paula McCourt1, Hsing-Yin Liu1, Josie E Parker2
1The Institute of Metabolic Disorders, Genesis Biotechnology Group, Hamilton, New Jersey 08691.
Abstract:
Candida albicans is an opportunistic fungus responsible for the majority of systemic fungal infections. Multiple factors contribute to C. albicans pathogenicity. C. albicans strains lacking CaArv1 are avirulent. Arv1 has a conserved Arv1 homology domain (AHD) that has a zinc-binding domain containing two cysteine clusters. Here, we explored the role of the CaAHD and zinc-binding motif in CaArv1-dependent virulence. Overall, we found that the CaAHD was necessary but not sufficient for cells to be virulent, whereas the zinc-binding domain was essential, as Caarv1/Caarv1 cells expressing the full-length zinc-binding domain mutants, Caarv1C3S and Caarv1C28S, were avirulent. Phenotypically, we found a direct correlation between the avirulence of Caarv1/Caarv1, Caarrv1 , Caarv1 , and Caarv1 cells and defects in bud site selection, septa formation and localization, and hyphal formation and elongation. Importantly, all avirulent mutant strains lacked the ability to maintain proper sterol distribution. Overall, our results have established the importance of the AHD and zinc-binding domain in fungal invasion, and have correlated an avirulent phenotype with the inability to maintain proper sterol distribution.
Insights
The Arv1 homology domain (AHD) and its zinc-binding motif are crucial for Candida albicans virulence. Defects in these domains prevent proper sterol distribution, leading to avirulence in this opportunistic fungus.
Area of Science:
- Medical Mycology
- Molecular Biology
- Cell Biology
Background:
- Candida albicans is a major cause of systemic fungal infections.
- Virulence of C. albicans depends on multiple factors, including the CaArv1 protein.
- CaArv1 contains a conserved Arv1 homology domain (AHD) with a zinc-binding motif.
Purpose of the Study:
- To investigate the role of the CaAHD and zinc-binding motif in CaArv1-dependent virulence.
- To determine the necessity and sufficiency of the CaAHD for virulence.
- To assess the impact of zinc-binding domain mutations on C. albicans pathogenicity.
Main Methods:
- Genetic manipulation of C. albicans to create strains lacking CaArv1 or expressing mutated CaArv1.
- Phenotypic analysis of mutant strains, including assessment of virulence, bud site selection, septa formation, and hyphal development.
- Evaluation of sterol distribution in wild-type and mutant strains.
Main Results:
- The CaAHD is necessary but not sufficient for C. albicans virulence.
- Mutations in the zinc-binding domain (Caarv1C3S and Caarv1C28S) render the fungus avirulent.
- Avirulent mutants exhibit defects in bud site selection, septa formation, and hyphal growth.
- All avirulent strains demonstrated impaired sterol distribution.
Conclusions:
- The AHD and zinc-binding domain of CaArv1 are essential for C. albicans virulence.
- Proper sterol distribution is critical for C. albicans pathogenicity.
- The study establishes a link between CaArv1 function, sterol homeostasis, and fungal virulence.
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