Unknown,Transcriptomics,Genomics,Proteomics

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Comparative Genome Hybridization between Candida albicans and Candida dubliniensis and their derivatives

ABSTRACT: Total genomic DNAs of CAKS20, CdUM4b, the pseudotetraploid hybrid strain CACD1 and the progeny hybrid strain Pre3 were isolated. In an attempt to determine the number of copies of individual chromosomes, we used comparative genome hybridization (CGH) analysis using specially designed whole genome microarrays representing C. albicans and C. dubliniensis genomes. The arrays contained 60 mer probes that fell in three categories either C. albicans specific or C. dubliniensis specific or common to both. The specific probes were expected to work as internal controls when hybridized to parental genomic DNA alone. The arrays were individually hybridized to Cy3-labeled genomic DNA from either of C. albicans (CAKS20), C. dubliniensis (CdUM4b) parents or the somatic psudotetraploid hybrid strain, CACD1 and the pseudodiploid progeny hybrid Pre3. After experimentally determining species-specific probes and common probes, the ratios of intensity on the CACD1 hybrid array to those of either of the parents were calculated. The copy number was determined for the entire chromosomes by comparing individual probe intensities from the pseudotetraploid to either diploid parent genome DNA intensities. Agilent one-color CGH experiment,Organism: Human ,Agilent-24598 Genotypic designed C.albicans and C. dubliniensis Cross Species CGH Microarray 4x180k , Labeling kit: Agilent Genomic DNA labeling Kit (Part Number: 5190-0453)

ORGANISM(S): Candida albicans

SUBMITTER: Kaustuv Sanyal

PROVIDER: E-GEOD-24269 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Although Candida albicans and Candida dubliniensis are most closely related, both species significantly behave differently with respect to morphogenesis and virulence. In order to gain further insight into the divergent routes for morphogenetic adaptation in both species, we investigated qualitative along with quantitative differences in the transcriptomes of both organisms by cDNA deep sequencing. Following genome-associated assembly of sequence reads we were able to generate experimentally verified databases containing 6016 and 5972 genes for C. albicans and C. dubliniensis, respectively. About 95% of the transcriptionally active regions (TARs) contain open reading frames while the remaining TARs most likely represent non-coding RNAs. Comparison of our annotations with publically available gene models for C. albicans and C. dubliniensis confirmed approximately 95% of already predicted genes, but also revealed so far unknown novel TARs in both species. Qualitative cross-species analysis of these databases revealed in addition to 5802 orthologs also 399 and 49 species-specific protein coding genes for C. albicans and C. dubliniensis, respectively. Furthermore, quantitative transcriptional profiling using RNA-Seq revealed significant differences in the expression of orthologs across both species. We defined a core subset of 84 hyphal-specific genes required for both species, as well as a set of 42 genes that seem to be specifically induced during hyphal morphogenesis in C. albicans. Species specific adaptation in C. albicans and C. dubliniensis is governed by individual genetic repertoires but also by altered regulation of conserved orthologs on the transcriptional level. We investigated qualitative along with quantitative differences in the transcriptomes of both organisms by cDNA deep sequencing. In a first step, we reevaluated the in silico predicted gene models by collecting experimental data using FLX - technology for sequencing strand-specific and normalized cDNA libraries derived from blastospores and hyphae. In the second step, quantitative RNA-Seq (GAIIX) was applied to C. albicans hyphal cells and C. dubliniensis blastospore and hyphal cells to complement reevaluation of the gene models with FLX data as well as to measure differential gene expression across the species with two biological replicates.

Project description:Candida albicans and Candida dubliniensis are closely related species displaying differences in virulence and genome content, therefore providing potential opportunities to identify novel C. albicans virulence genes. C. albicans gene arrays were used for comparative analysis of global gene expression in the two species in reconstituted human oral epithelium (RHE). C. albicans (SC5314) showed upregulation of hypha-specific and virulence genes within 30 min postinoculation, coinciding with rapid induction of filamentation and increased RHE damage. C. dubliniensis (CD36) showed no detectable upregulation of hypha-specific genes, grew as yeast, and caused limited RHE damage. Several genes absent or highly divergent in C. dubliniensis were upregulated in C. albicans. One such gene, SFL2 (orf19.3969), encoding a putative heat shock factor, was deleted in C. albicans. ÎÎsfl2 cells failed to filament under a range of hypha-inducing conditions and exhibited greatly reduced RHE damage, reversed by reintroduction of SFL2 into the ÎÎsfl2 strain. Moreover, SFL2 overexpression in C. albicans triggered hyphal morphogenesis. Although SFL2 deletion had no apparent effect on host survival in the murine model of systemic infection, ÎÎsfl2 strain-infected kidney tissues contained only yeast cells. These results suggest a role for SFL2 in morphogenesis and an indirect role in C. albicans pathogenesis in epithelial tissues. Global gene expression in Candida albicans SC5314 or Candida dubliniensis CD36 on reconstituted human oral epithelium (RHE) 90 min postinoculation. Gene expression on RHE was compared to gene expression on polycarbonate filter (PCF; used as RHE support matrix) 90 min postinoculation. Performed in two biological replicates with reciprocal dye swaps for each biological replicate. Gene expression of Candida cells on RHE was normalized to gene expression in reference control (PCF); log2 ratios were calculated by dividing spot intensity of experimental by that of the reference control.

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Comparative Genome Hybridization between Candida albicans and Candida dubliniensis and their derivatives

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