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Transcriptomic profiling of dimorphic growth in two Ustilaginomycotina species

ABSTRACT: In this study, we performed comparative transcriptomics to understand how mechanisms for fungal dimorphism are similar to the model species Ustilago maydis. To achieve this, we selected two non-model species from Ustilaginomycotina, Meira miltonrushii and Tilletiopsis washingtonensis, for transcriptomic profiling. Different culture conditions were used to have these two fungi display yeast growth (Holliday's minimal media with 1% glucose as a carbon source) and filamentous growth (Holliday's minimal media with 1% Tween as a carbon source). Then, we performed transcriptomic sequencing and analyses to determine which genes are differentially expressed in filamentous growth compared to yeast growth. We also incorporated public datasets from other two dimorphic species, U. maydis and Ophiostoma novo-ulmi, for comparison. We found that T. washingtonensis, despite being closely related to M. miltonrushii, is the most distant species in terms of the similarities of transcriptomic alteration, suggesting phylogenetic relatedness may not be concurrent with transcriptomic similarity under the same biological phenomenon. Functional enrichment analyses reveal that genes in cell energy production and conversion, amino acid transport and metabolism and cytoskeleton are groups of genes differentially expressed in common among the four species. Finally, although our comparative approach allows us to discover several candidate genes that may be involved in fungal dimorphism, we found very few commonly expressed genes that are know dimorphism genes characterized in U. maydis literature. This suggests the certain degree of species-specific mechanisms for dimorphic transition.

ORGANISM(S): Meira miltonrushii Tilletiopsis washingtonensis

PROVIDER: GSE154947 | GEO | 2021/06/09

REPOSITORIES: GEO

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Project description:Thermally dimorphic human fungal pathogens undergo a reversible program of cellular differentiation in response to their environment that is essential for infectivity and pathogenicity. In the soil, these organisms grow as highly polarized, multicellular hyphal filaments that produce infectious particles. When inhaled by a mammalian host, these cells switch to a unicellular yeast form that causes disease even in healthy hosts. Temperature is considered to be the primary environmental cue that promotes reversible cellular differentiation; however, a shift to a lower temperature in vitro induces filamentous growth in an inefficient and asynchronous manner. In a search for other signals that regulate morphogenesis, we considered the monosaccharide N-acetylglucosamine (GlcNAc), which is a major component of microbial cell walls and is ubiquitous in the environment. GlcNAc was a potent and specific inducer of the yeast-to-filament transition in two thermally dimorphic fungi, Histoplasma capsulatum and Blastomyces dermatitidis. Micromolar concentrations of GlcNAc induced a robust morphological transition of H. capsulatum after temperature shift, indicating that fungal cells sense GlcNAc to promote filamentation. The synchronous morphologic transition stimulated by low temperature and GlcNAc allowed us to examine the temporal regulation of the transcriptome during morphogenesis to reveal candidate genes involved in establishing the filamentous growth program. Through this analysis, we identified two genes encoding GlcNAc transporters, NGT1 and NGT2, that were necessary for H. capsulatum cells to robustly filament in response to GlcNAc. Unexpectedly, NGT1 and NGT2 were important for efficient H. capsulatum yeast-to-filament conversion in standard glucose medium, suggesting that Ngt1 and Ngt2 monitor endogenous levels of GlcNAc to control multicellular filamentous growth in response to temperature. Overall, our work indicates that GlcNAc functions as a highly conserved cue of morphogenesis in fungi, which further enhances the significance of this ubiquitous sugar in cellular signaling in eukaryotes. For each time-course sample, cDNA was coupled to Cy5 and a reference cDNA pool was made by combining RNA from t = 0 and all late time course samples, which was coupled to Cy3. For end point microarray experiments (i.e., established yeast samples compared to established filamentous samples), G217B yeast cDNA was coupled to Cy5 and filament cDNA was coupled to Cy3.

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Transcriptomic profiling of dimorphic growth in two Ustilaginomycotina species

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