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Regulation of the biofilm-specific genes circuitry in Candida albicans by a novel histone H3 variant

ABSTRACT: To understand the biological relevance of the role played by the HHT1 histone H3 variant in C. albicans, we performed a transcriptome analysis of the mutant by global gene expression array analysis. We analyzed gene expression profile of the mutant in strains LR107 and LR108 (hht1∆/hht1∆) and the parent wild type (SC5314) strain grown in YPD liquid medium at 30°C. In the microarray analysis, a total of 1222 genes were found to be expressed significantly different (fold change < 1.5 at p-value < 0.05) between wild type and two hht1 null mutants. Out of the 1222 differentially expressed genes, 670 genes were up-regulated whereas 552 genes were down-regulated. Gene ontology (GO) analysis of up-regulated and down-regulated genes suggests that biofilm, Hap43, transcription, filamentation, white-opaque/mating and invasive growth are the major pathways altered in minor H3 null mutant. Transcription profiling in planktonic YPD condition suggests that minor H3 represses biofilm genes in the planktonic mode of growth. Therefore we performed a genome-wide expression array study after growing the wild type strain and hht1/hht1 mutant strains in Spider induced biofilm condition. This experiment was carried out with two biological replicates of hht1 null mutants. The biofilm induced microarray data revealed that a total of 1010 genes were differentially expressed (at p-value <0.05 and fold change <1.5) between the wild type and hht1 mutants. Out of 1016 differentially expressed genes, 572 genes were up-regulated whereas 444 genes were down-regulated.

ORGANISM(S): Candida albicans

PROVIDER: GSE72824 | GEO | 2016/09/08

SECONDARY ACCESSION(S): PRJNA295172

REPOSITORIES: GEO

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Project description:Biofilm formation on medically implanted devices by Candida albicans poses a significant clinical challenge. Here we compared biofilm-associated gene expression in two clinical C. albicans isolates, SC5314 and WO-1, to identify shared gene regulatory responses that may be functionally relevant. Among the 50 genes most highly expressed in biofilms relative to planktonic (suspension-grown) cells, we were able to recover insertion mutations in 25 genes. We observed that 20 of the 25 mutants have altered biofilm-related properties, including cell-substrate adherence, cell-cell signaling, and azole susceptibility. We focused on the most highly up-regulated gene in biofilms, RHR2, which specifies the glycerol biosynthetic enzyme glycerol-3-phosphate phosphatase. Glycerol is 5-fold more abundant in biofilm cells than planktonic cells, and an rhr2D/D strain accumulates 2-fold less biofilm glycerol than the wild type. Under in vitro growth conditions, the rhr2D/D mutant has reduced biofilm biomass and reduced adherence to silicone. The rhr2D/D mutant is severely defective in biofilm formation in vivo, in a rat catheter infection model. Expression profiling of the rhr2D/D mutant indicates that it has reduced expression of cell surface adhesin genes ALS1, ALS3, and HWP1, as well as a large fraction of all other biofilm up-regulated genes. Reduced adhesin expression is the cause of the rhr2D/D mutant biofilm defect, because overexpression of ALS1, ALS3, or HWP1 restores biofilm formation ability to the mutant in vitro and in vivo. Our findings indicate that internal glycerol has a regulatory role in biofilm gene expression, and that adhesin genes are among the main functional Rhr2-regulated genes. Gene expression profiles, in duplicate; (1) for biofilm vs. planktonic growth conditions for the two wild-type clinical isolates of Candida albicans (SC5314 and WO1-white/WO1-opaque), and (2) for rhr2M-NM-^T/M-NM-^T mutant and complemented strain, via RNA-deep sequencing using Illumina GA2 and HiSeq2000 platforms, respectively

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Regulation of the biofilm-specific genes circuitry in Candida albicans by a novel histone H3 variant

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