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Published on: March 19, 2019
Isolation and expression of the gene encoding mitochondrial ADP/ATP carrier (AAC) from the pathogenic yeast Candida
M Nebohácová1, M Mentel, J Nosek
1Department of Biochemistry, Faculty of Sciences, Comenius University, Bratislava, Slovakia.
Abstract:
A gene homologous to Saccharomyces cerevisiae AAC genes coding for mitochondrial ADP/ATP carriers has been cloned from the pathogenic yeast Candida parapsilosis. A probe obtained by PCR amplification from C. parapsilosis DNA, using primers derived from the conserved transmembrane region of yeast ADP/ATP carriers, was used for screening of the C. parapsilosis genomic library. The cloned gene was sequenced and found to encode a polypeptide of 303 amino acids that shows homology with other yeast and fungal mitochondrial ADP/ATP carriers. The gene was designated CpAAC1 and was able to complement the growth phenotypes of S. cerevisiae double deletion mutant (Deltaaac2; Deltaaac3). The expression of the CpAAC1 gene was reduced under semi-anaerobic conditions and it was affected at normal aerobic conditions by the nature of carbon sources used for growth. Hybridization experiments indicate that C. parapsilosis possesses a single gene encoding a mitochondrial ADP/ATP carrier.
Insights
Researchers cloned the CpAAC1 gene from Candida parapsilosis, which codes for mitochondrial ADP/ATP carriers. This gene complements yeast mutants, showing its functional similarity and potential role in fungal energy metabolism.
Area of Science:
- Molecular Biology
- Mycology
- Biochemistry
Background:
- Mitochondrial ADP/ATP carriers are crucial for cellular energy homeostasis.
- The pathogenic yeast Candida parapsilosis requires functional mitochondrial carriers for survival.
Purpose of the Study:
- To clone and characterize a gene encoding a mitochondrial ADP/ATP carrier from Candida parapsilosis.
- To investigate the functional complementation and expression patterns of the identified gene.
Main Methods:
- Polymerase Chain Reaction (PCR) amplification using conserved primers.
- Screening of a Candida parapsilosis genomic library.
- Gene sequencing and functional complementation assays in Saccharomyces cerevisiae.
- Analysis of gene expression under varying growth conditions.
Main Results:
- A gene, designated CpAAC1, was successfully cloned and sequenced, encoding a 303-amino acid polypeptide homologous to known yeast ADP/ATP carriers.
- CpAAC1 demonstrated functional complementation of Saccharomyces cerevisiae double deletion mutants lacking essential ADP/ATP carriers.
- CpAAC1 expression was downregulated under semi-anaerobic conditions and influenced by carbon source availability during aerobic growth.
- Hybridization studies confirmed the presence of a single gene for mitochondrial ADP/ATP carrier in Candida parapsilosis.
Conclusions:
- The cloned CpAAC1 gene encodes a functional mitochondrial ADP/ATP carrier in Candida parapsilosis.
- CpAAC1 plays a role in energy metabolism and its expression is regulated by oxygen availability and nutrient sources.
- This study provides insights into the molecular mechanisms of energy transport in pathogenic fungi.
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