Project description:Transcriptional profiling of C. lusitaniae a/alpha WT cells on 0.37% PDA (potato dextrose agar) for 30 minutes (m), 2 hours (h), 4h, 8h, 12h, 18h, 24, and 30h hybridized against WT a/alpha cells in non-meiosis inducing conditions YPD (yeast peptone dextrose) for 2h Two condition experiment: a/alpha WT on PDA vs. a/alpha WT on YPD, 4 biological replicates
Project description:Transcriptional profiling of C. lusitaniae a and alpha cells mixed on 0.37% PDA (potato dextrose agar) for 0 hours (h) ,4h, and 12n hybridized against WT cells on YPD for 4 hours and also a and alpha ime2 deletion mutants mixed on 0.37% PDA for 4 hours hybridized against WT cells on YPD (yeast peptone dextrose) for 4h Two condition experiment: WT a and alpha cells on 0.37% PDA and a and alpha ime2 deletion mutants mixed on 0.37% PDA. Three biological replicates
Project description:Transcriptional profiling of C. lusitaniae a/alpha ste12Δ/ste12Δ deletion mutants on .37% PDA (potato dextrose agar) media for 0 hours (h), 12h, 28h, and 24 hours hybridized against WT a/alpha cells on YPD (yeast peptone dextrose) media for 2h Two condition experiment: 2N (a/alpha) ste12Δ/ste12Δ (CAY4240) on PDA vs. 2N WT (RSY432) on YPD, 1 biological replicate
Project description:Transcriptional profiling of C. lusitaniae WT a and WT alpha cells on 0.37% PDA (potato dextrose agar) for 30 minutes (m), 2 hours (h), or 24h hybridized against WT a and WT alpha cells in YPD (yeast peptone dextrose) for 2h WT a or WT alpha cells were incubated on PDA medium for 30 minutes, 2 hours, or 24 hours and hybridized against WT a or alpha cells (respectively) incubated on YPD for 2 hours, 2 biological replicates
Project description:Transcriptional profiling of C. lusitaniae ime2Δ/ime2Δ a/alpha WT cells on 0.37% PDA (potato dextrose agar) for 0 hours, 12h, 18h and 24h hybridized against WT a/alpha cells in YPD (yeast peptone dextrose) for 4h Two condition experiment: ime2Δ/ime2Δ a/alpha WT on PDA vs. WT a/alpha on YPD, 3 biological replicates
Project description:Transcriptional profiling of a-type wor1 deleted cells and mixed a-type and alpha-type opaque cells under in vitro biofilm-forming conditions. Specifically, they were grown for two days at room temperature in a 12-well poly-styrene plate containing 1 ml of Lee's + Glucose liquid media. Samples were hybridized against a universal mixed reference sample of a-type cells in white and opaque states grown in Spider liquid media. 2 condition experiment: white wor1-deletion mutant a-type cells, opaque mixed a-type and alpha-type cells; two biological replicates each.
Project description:Transcriptional profiling of C. tropicalis a/alpha cells (CAY1511) in white state, opaque state, overexpressing Wor1, or wor1 mutants hybridized against a universal mixed reference sample from all 4 states. 4 condition experiment: white, opaque, tdh3-wor1, Δ/Δwor1; 4 biological replicates of each
Project description:Act1-Wor1 strain (the opaque strain CAY2903 with constitute WOR1 expression), wildtype white (a/a RBY717); Opaque hyphal formation in liquid LP and SOR compared to SCD; white cells in the same conditions as control. 6 condition experiment: opaque and white cells in LP, SOR and SCD. Used the pool of all conditions as reference.
Project description:Candida albicans can stochastically switch between two phenotypes, white and opaque. Opaque cells are the sexually competent form of C. albicans and therefore undergo efficient polarized growth and mating in the presence of pheromone. In contrast, white cells cannot mate, but are induced - under a specialized set of conditions - to form biofilms in response to pheromone. In this work, we compare the genetic regulation of such "pheromone-stimulated" biofilms with that of "conventional" C. albicans biofilms. In particular, we examined a network of six transcriptional regulators (Bcr1, Brg1, Efg1, Tec1, Ndt80, and Rob1) that mediate conventional biofilm formation for their potential roles in pheromone-stimulated biofilm formation. We show that four of the six transcription factors (Bcr1, Brg1, Rob1, and Tec1) promote formation of both conventional and pheromone-stimulated biofilms, indicating they play general roles in cell cohesion and biofilm development. In addition, we identify the master transcriptional regulator of pheromone-stimulated biofilms as C. albicans Cph1, ortholog of Saccharomyces cerevisiae Ste12. Cph1 regulates mating in C. albicans opaque cells, and here we show that Cph1 is also essential for pheromone-stimulated biofilm formation in white cells. In contrast, Cph1 is dispensable for the formation of conventional biofilms. The regulation of pheromone- stimulated biofilm formation was further investigated by transcriptional profiling and genetic analyses. These studies identified 206 genes that are induced by pheromone signaling during biofilm formation. One of these genes, HGC1, is shown to be required for both conventional and pheromone-stimulated biofilm formation. Taken together, these observations compare and contrast the regulation of conventional and pheromone-stimulated biofilm formation in C. albicans, and demonstrate that Cph1 is required for the latter, but not the former. 4 condition experiment: white and opaque cells in planktonic and pheromone-induced biofilm conditions with and without alpha pheromone. WT strain (P37005), the tec1 mutant strain and the cph1 mutant strain
Project description:Sexual reproduction can promote genetic diversity in eukaryotes, and yet many pathogenic fungi have been labeled as obligate asexual species. It is becoming increasingly clear, however, that cryptic sexual programs may exist in some species, and that efficient mating requires the necessary developmental switch to be triggered. In this study we investigate Candida tropicalis, an important human fungal pathogen that has been reported to be asexual. Significantly, we demonstrate that C. tropicalis uses a phenotypic switch to regulate a cryptic program of sexual mating. Thus, diploid a and α cells must undergo a developmental transition to the mating-competent form, and only then does efficient cell-cell conjugation take place resulting in the formation of stable a/α tetraploids. We show that both the phenotypic switch and sexual mating depend on the conserved transcriptional regulator Wor1, which is regulated by temperature in other fungal species. In contrast, C. tropicalis mating occurs efficiently at both 25 °C and 37 °C, suggesting that it could occur in the mammalian host and have direct consequences for the outcome of an infection. Transcriptional profiling further reveals that ≈400 genes are differentially expressed between the two phenotypic states, including the regulatory factor Wor1. Taken together, our results demonstrate that C. tropicalis has a unique sexual program, and that entry to this program is controlled via a Wor1-mediated, metastable switch. These observations have direct implications for the regulation and evolution of cryptic sexual programs in related fungal pathogens. 4 biological replicates of both the white (CAY1504) and opaque (CAY2275) states of C. tropicalis a cells are included on this array. All are hybridized against a universal reference sample, which consists of the combined RNA from all 8 replicates used on this array.