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In vivo RNA sequencing reveals a crucial role of Fus3-Kss1 MAPKpathway in Candida glabrata pathogenicity

ABSTRACT: Candida glabrata is an important and increasingly common pathogen of humans, particularly in immunocompromised hosts. Despite this, little is known about how this fungus causes disease. Here, we applied RNA sequencing and an in vivo invasive infection model to identify the attributes that allow this organism to infect hosts. Fungal transcriptomes show a dramatic increase in the expression of Fus3 and Kss1, two mitogen-activated protein kinases (MAPKs), during invasive infection. We further demonstrate that they are both highly induced under a combination of serum and high CO2 condition. Deletion of both FUS3 and KSS1, but neither gene alone, results in a reduced fungal burden in organs, as well as in gastrointestinal tract in the DSS (Dextran Sulfate Sodium)-induced colitis model. Similarly, the defect in persistence in macrophages and attenuated adhesion to epithelial cells are observed when FUS3 and KSS1 are both disrupted. The fus3 kss1 double mutant also displays defects in the induction of virulence attributes such as genes required for iron acquisition and adhesion, and in the anti-fungal drug tolerance. The putative downstream transcription factors Ste12(1), Ste12(2), Tec1 and Tec2 are found to be involved in the regulation of these virulence attributes. Collectively, our study indicates that an evolutionary conserved MAPK pathway which regulates mating and filamentous growth in Saccharomyces cerevisiae, is critical for C. glabrata pathogenicity.

ORGANISM(S): Nakaseomyces glabratus

PROVIDER: GSE279281 | GEO | 2024/10/16

REPOSITORIES: GEO

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Project description:The FUSCA3 (FUS3) transcription factor is considered to be a master regulator of seed maturation because a wide range of seed maturation events are impaired in its defective mutant. To comprehensively identify genes under the control of FUS3, two types of microarray experiments were performed. First, transgenic plants in which FUS3 expression could be induced by the application of estrogen (ESTR) were used to identify the genes up-regulated in young seedlings of Arabidopsis in response to ectopic expression of FUS3. Second, transcriptomes were compared between fus3 mutant and wild type developing seeds. Combining the results of these experiments identified genes under relatively immediate and robust control of FUS3 during seed development. The analyses expanded the range of types of genes under the control of FUS3. The genes positively controlled by FUS3 are not confined to previously known seed maturation-related genes and include those for production of secondary metabolites such as glucosinolates, phenylpropanoids and falvonoids, and primary metabolism such as photosynthesis and fatty acid biosynthesis. Furthermore, several different patterns were identified in the manner of ectopic activation by FUS3 including the kinetics and abscisic acid (ABA) requirement of downstream genes depending on their natures of developmental regulation, suggesting mechanistic diversity of gene regulation by FUS3. This Series contains two different types of microarray experiments. First, seven-day-old seedlings of the ESTR-inducible FUS3 transgenic plants (ER-FUS3) were cultured for 3, 6, 12, 24, 36 or 48 h in a standard liquid medium or in that containing ESTR, ABA or both. RNA was prepared from each time point sample of each treatment and used to prepare Cy5-labeled cRNA, which was subsequently probed by DNA array with the reference Cy3-labeled cRNA prepared from the time zero untreated seedlings. Two biological replicates per treatment. Second, transcriptomes were compared between developing seeds of the wild-type (Col-0) and the fus3-3 mutant. This experiment contains two seed developmental stages, 8 days after flowering (DAF; bent cotyledon stage) and at 12 DAF (green cotyledon stage). Sample pairs in three biological replicates were analyzed by the two-color method. Dye-swapped hybridizations were performed in every replicate.

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In vivo RNA sequencing reveals a crucial role of Fus3-Kss1 MAPKpathway in Candida glabrata pathogenicity

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