Project description:Comparative analysis of genome wide binding profile of Ncb2 in azole sensitive (AS, Gu4) and azole resistant (AR, Gu5) clinical isolates of Candida albicans. The goal was to study the role of Ncb2 in acquisition of drug resistance by comparing the binding profiles of Ncb2 in both the isolates.
Project description:RNA sequencing was performed on Candida albicans clinical isolates that display normal (isolates: 3560, 3605, 3609, 4108, 4259) or aberrant (isolates: 3534, 3544, 3621, 3636, 4036) beta-glucan masking in response to lactate and hypoxia. Each clinical isolate was grown to exponential phase in GYNB under normoxic conditions and then exposed for 5 h to: (a) 1% lactate; (b) 0% lactate control; (c) hypoxia; or (d) normoxic control. Three independent experiments were performed for each clinical isolate.
Project description:Invasive fungal infections are important healthcare associated disease worldwide especially in intensive care units More recently, Candida auris a multidrug and potentially pan-resistant species has globally emerged as a new nosocomial pathogen, which has been already reported from at least 50 countries on six continents. Clinical studies showed that previously well-defined phylogenetic C. auris clades display significant differences regarding their pathogenicity, virulence, metabolism and susceptibility profile to traditional antifungal therapies. Based on epidemiological data, isolates belonging to the South Asian clade show the highest ratio of resistance to fluconazole (97%), amphotericin B (47%) and this clade involves the highest number of multidrug resistant isolates (45%), which compromise the efficacy of applied antifungal therapy. In the past decade, a new broad-spectrum antifungal drug, isavuconazole (ISA), has been introduced into clinical practice. ISA is primarily approved for the treatment of invasive aspergillosis and mucormycosis, and currently, there are no available recommendations for the therapy of invasive Candida infections. In our previous study, we reported different ISA susceptibility profiles between isolates belonging to South Asian lineage. However, the global transcriptional - even isolate specific - response remained unresolved. Therefore, our study aimed to reveal those molecular events, which are associated with ISA exposure using high throughput RNA sequencing (RNAseq).
Project description:Rme1, a conserved transcription factor among members of the ascomycete lineage, regulates meiosis and pseudohyphal growth in baker’s yeast. The genome of the meiosis-defective fungal pathogen Candida albicans encodes a Rme1 homolog, which we previously mapped within a transcriptional circuitry that controls hyphal growth. To delineate a possible role of Rme1 in C. albicans morphogenesis, we combined genome-wide expression and location analyses of Rme1. Strikingly, Rme1 bound upstream and activated the expression of markers of chlamydosporulation, a process leading to formation of large, spherical, thick-walled cells during nutrient starvation. RME1 deletion abolished chlamydosporulation in three chlamydospore-forming Candida species, whereas its overexpression bypassed the requirement for chlamydosporulation cues and regulators, indicating that Rme1 is central to chlamydospore development. Moreover, RME1 expression levels correlated with chlamydosporulation efficiency among clinical isolates, further highlighting Rme1 importance in this process. Interestingly, RME1 displayed a biphasic pattern of expression, with a first phase independent of Rme1 function and dependent on chlamydospore-inducing cues, and a second phase depending upon Rme1 function and independent of chlamydospore-inducing cues. We suggest that Rme1 function spans from the regulation of meiosis in sexual yeasts to the control of an epigenetic switch necessary for asexual spore formation in meiosis-defective Candida species.