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Validation of Cdc68p as a novel antifungal target
Ed T Buurman1, Weidong Jiang, Melissa McCoy
1Anadys Pharmaceuticals Ltd., 9050 Camino Santa Fe, San Diego, CA 92121, USA. Ed.Buurman@astrazeneca.com
Abstract:
Candida albicans is the main cause of systemic fungal infections for which there is an urgent need for novel antifungal drugs. The CP (Cdc68p-Pob3p) complex, which is involved in transcription elongation, was evaluated as a putative antifungal target. In order to predict the consequences of inhibition of this complex, the largest CP subunit in Saccharomyces cerevisiae, Cdc68p, was the first novel target to be tested in GATE, a recently described, quantitative target inactivation system. Depletion of the cell's pool of Cdc68p led to rapid cell death. Subsequently, the C. albicans orthologue of CDC68, CaCDC68, was cloned. Attempts to disrupt both alleles were unsuccessful, thus suggesting an essential role of CaCDC68 in this fungus also. Furthermore, CDC68 was proven to be present in Neurospora crassa and Aspergillus nidulans, thus suggesting that the CP complex is widespread among fungi and could serve as a broad range antifungal target. Analysis of Cdc68p and Pob3p sequences indicated significant structural differences between fungal CP complexes and those present in higher eukaryotes. These results predict that, in principle, fungal-specific ligands of CP complexes could be identified that could subsequently serve as chemical starting points towards the development of new antifungal therapeutic agents.
Insights
The Cdc68p-Pob3p (CP) complex is a potential antifungal target. Inhibiting this essential fungal complex could lead to new treatments for systemic fungal infections.
Area of Science:
- Mycology
- Molecular Biology
- Drug Discovery
Background:
- Candida albicans causes dangerous systemic fungal infections, necessitating new antifungal drugs.
- The Cdc68p-Pob3p (CP) complex, crucial for transcription elongation, is explored as a novel antifungal target.
Purpose of the Study:
- To evaluate the CP complex as a potential antifungal target.
- To investigate the essentiality of the CP complex in Candida albicans and related fungi.
Main Methods:
- Utilized the GATE system to inactivate Cdc68p in Saccharomyces cerevisiae.
- Cloned and attempted to disrupt the CaCDC68 gene in Candida albicans.
- Performed sequence analysis of fungal CP complexes and compared them to higher eukaryotes.
Main Results:
- Depletion of Cdc68p in S. cerevisiae caused rapid cell death, indicating its essential role.
- CaCDC68 was found to be essential in Candida albicans, as biallelic disruption was unsuccessful.
- The CDC68 gene is conserved in other fungi like Neurospora crassa and Aspergillus nidulans, suggesting the CP complex is a widespread fungal target.
- Significant structural differences were identified between fungal and higher eukaryotic CP complexes.
Conclusions:
- The CP complex is essential in fungi and represents a promising broad-range antifungal target.
- Structural differences suggest the possibility of developing fungal-specific inhibitors for new antifungal therapies.