Related Experiment Video
Updated: Jul 16, 2026
Whole Genome Sequencing of Candida glabrata for Detection of Markers of Antifungal Drug Resistance
Published on: December 28, 2017
The early transcriptional and post-transcriptional responses to fluconazole in sensitive and resistant Candida
Irene Stevens1, Fitz Gerald Silao2, Susanne Huch1
1Science for Life Laboratory (SciLifeLab), Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Solna, Sweden.
Abstract:
Candida albicans is a leading cause of fungal infections in immunocompromised patients. Management of candidemia relies on a few antifungal agents, with fluconazole being first line therapy. The emergence of fluconazole-resistant strains highlights the pressing need to improve our molecular understanding of the drug response mechanisms. By sequencing the 5'P mRNA degradation intermediates, we establish that co-translational mRNA decay occurs in C. albicans and characterize how in vivo 5´-3´ exonuclease degradation trails the last translating ribosome. Thus, the study of the 5' Phosphorylated mRNA degradome (5PSeq) offers a simple and affordable way to measure ribosome dynamics and identify codon specific ribosome stalls in response to drugs and amino acid deprivation. Building upon this, we combine RNA-Seq and 5PSeq to study the early response of sensitive and resistant C. albicans isolates to fluconazole. Our results highlight that transcriptional responses, rather than changes in ribosome dynamics, are the main driver of Candida resistance to fluconazole.
Insights
The study reveals that Candida albicans resistance to fluconazole primarily stems from transcriptional changes, not altered ribosome dynamics. This finding offers new insights into antifungal drug resistance mechanisms.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Candida albicans causes significant fungal infections, particularly in immunocompromised individuals.
- Fluconazole is the primary treatment for candidemia, but resistance is a growing concern.
- Understanding fluconazole resistance mechanisms is crucial for developing effective therapies.
Purpose of the Study:
- To investigate the molecular mechanisms underlying fluconazole resistance in Candida albicans.
- To characterize mRNA decay pathways and ribosome dynamics in response to fluconazole.
- To differentiate the roles of transcriptional changes versus ribosome dynamics in drug resistance.
Main Methods:
- Utilized 5' Phosphorylated mRNA degradome sequencing (5PSeq) to analyze mRNA decay intermediates.
- Applied RNA-Sequencing (RNA-Seq) to assess gene expression changes.
- Combined RNA-Seq and 5PSeq to study early fluconazole responses in sensitive and resistant C. albicans strains.
Main Results:
- Established that co-translational mRNA decay occurs in C. albicans, with degradation trailing the ribosome.
- Demonstrated that 5PSeq can measure ribosome dynamics and detect codon-specific ribosome stalls.
- Identified transcriptional responses as the primary driver of fluconazole resistance in C. albicans, overshadowing changes in ribosome dynamics.
Conclusions:
- Transcriptional alterations are the key mechanism conferring fluconazole resistance in Candida albicans.
- 5PSeq is a valuable tool for studying ribosome dynamics and drug responses.
- Further research into transcriptional regulation could yield novel antifungal strategies.
