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Cryptococcus neoformans var. neoformans

ABSTRACT: Cryptococcus neoformans sexual reproduction is controlled by a quorum sensing peptide

PROVIDER: PRJNA368839 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR5204780_1.fastq.gz	Fastqsanger.gz
	SRR5204780_2.fastq.gz	Fastqsanger.gz
	SRR5204781_1.fastq.gz	Fastqsanger.gz
	SRR5204781_2.fastq.gz	Fastqsanger.gz
	SRR5204782_1.fastq.gz	Fastqsanger.gz

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Cryptococcus neoformans sexual reproduction is controlled by a quorum sensing peptide

Project description:Mating pheromone is the best-known fungal paracrine signal whose essentiality in determining sexual reproduction has been recognized since six decades ago. We demonstrate that the pheromone cannot induce mating response during unisexual mating due to the lack of a compatible receptor from the same unisexual population. We identify a quorum sensing (QS) peptide Qsp1 in place of pheromone as a specific intercellular signal directing Cryptococcus unisexual reproduction. A typical zinc finger regulator Cqs2 as the master regulator of QS signaling induces unisexual reproduction in response to Qsp1. Collectively, these data demonstrate that QS activation rather than pheromone induction as the pivotal social control mechanism underlies unisexual reproduction in C .neoformans.

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Time-course microarray study of Cryptococcus neoformans var. neoformans (JEC20xJEC21) sexual development

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Project description:Time-course microarray study of Cryptococcus neoformans var. neoformans (JEC20xJEC21) sexual development

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Time-course microarray study of Cryptococcus neoformans var. neoformans (JEC20xJEC21) sexual development

Project description:To identify the genome-wide transcriptional changes that occur throughout sexual developmen of C. neoformans, we conducted a time-course microarray experiment spanning six developmental stages to generate a temporal expression pattern for each known gene. C. neoformans a x M-NM-1 co-cultures were grown under sexual development conditions (V8 agar), and RNA was harvested at 0.5, 6, 12, 24, 48, and 72 hours post-mixing. Time points were chosen based on microscopic identification of cell types of interest: fusants, filaments (early and late), basidia, and spores (Figure 1B). Microarray hybridizations were conducted in a loop design, with each sample serving as a reference for the following time point in the experiment (e.g. 0.5 hours vs. 6 hours, 6 hours vs. 12 hours, etc). The experiment spanned six time points of development: 0.5 hours, 6 hours, 12 hours, 24 hours, 48 hours and 72 hours. Hybridizations were conducted in a loop design, with each sample serving as the reference for the consecutive time point - resulting in 5 experimental comparisons: .5hr vs 6 hr, 6 hrs vs 12 hrs, 12 hrs vs 24 hrs, 24 hrs vs 48 hrs, and 48 hrs vs 72 hrs. Each comparison consistent of duplicate hybridizations with dye swaps. Arrays contain duplicate genomes, resulting in eight-fold coverage within each comparison. (total of 20 hybridizations)

2012-04-30 | E-GEOD-36977 | biostudies-arrayexpress

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Project description:Uniparental reproduction in Cryptococcus neoformans

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Coordinate genomic association of transcription factors controlled by an imported quorum sensing peptide in Cryptococcus neoformans

Project description:Coordinate genomic association of transcription factors controlled by an imported quorum sensing peptide in Cryptococcus neoformans

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Project description:Convergent evolution of peptide-based quorum sensing required for virulence in a eukaryotic pathogen

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Coordinate genomic association of transcription factors controlled by an imported quorum sensing peptide in Cryptococcus neoformans

Project description:Qsp1 is a secreted quorum sensing peptide required for virulence of the fungal meningitis pathogen Cryptococcus neoformans. Qsp1 functions to control cell wall integrity in vegetatively growing cells and also functions in mating. Rather than acting on a cell surface receptor, Qsp1 is imported to act intracellularly via the predicted oligopeptide transporter Opt1. Here, we identify a transcription factor network as a target of Qsp1. Using whole-genome chromatin immunoprecipitation, we find Qsp1 controls the genomic associations of three transcription factors to genes whose outputs are regulated by Qsp1. One of these transcription factors, Cqs2, is also required for the action of Qsp1 during mating, indicating that it might be a shared proximal target of Qsp1. Consistent with this hypothesis, deletion of CQS2 impacts the binding of other network transcription factors specifically to Qsp1-regulated genes. These genetic and genomic studies illuminate mechanisms by which an imported peptide acts to modulate eukaryotic gene expression.

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Project description:To identify the genome-wide transcriptional changes that occur throughout germination of C. neoformans spores, we conducted a time-course microarray experiment spanning six timepoints to generate a temporal expression pattern for each known gene. Spores were placed in rich medium and allowed to germinate for 10 hours, when they start to replicate as yeast. Each time point was flash frozen in liquid nitrogen and RNA was harvested for each time point at the same time. Microarray hybridizations were conducted in a reference pool design, where each time point was mixed together in equal amounts to make a reference pool sample. Then each time point was hybrdized against the reference pool. Characterization of the genes and pathways that are regulated during germination of this ubiquitous fungal pathogen will allow us to better understand how infectious spores resume vegetative growth, a process that likely is critical for interaction between C. neoformans and a host.

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Project description:Carbon dioxide (CO2) sensing, transport, and metabolism play a pivotal role in survival and proliferation of pathogenic microbes infecting human host from natural environments due to the drastic difference of CO2 levels. Carbonic anhydrases (CAs) are not only key CO2-metabolic enzymes catalyzing reversible interconversion between CO2 and bicarbonate (HCO3-), but also important CO2-signaling modulators. Cryptococcus neoformans that causes fatal fungal meningitis contains two functional CAs, Can1 and Can2, among which Can2 plays essential roles for growth and sexual differentiation of the pathogen. However, downstream genes and signaling network regulated by CAs have not been studied thus far. In this study, we constructed a C. neoformans strain where CAN2 expression is controlled by the CTR4 (copper transporter) promoter and elucidated its transcriptome patterns by using DNA microarray analysis to elucidate downstream target genes of Can2. Expectedly, the CAN2 promoter replacement strain showed growth defects in a CO2-dependent manner when CAN2 is repressed. The CA-transcriptome analysis discovered a number of Can2-dependent genes, including those involved in fatty acid biosynthesis (FAS1), sexual differentiation (GPB1), capsule structure organization (CAS3), iron-metabolism (CFO2), and a number of stress-regulated genes, including an oxidative stress-responsive Atf1 transcription factor, although a majority of them do not have any orthologs in other fungi. The present study revealed other roles of Atf1 in capsule and melanin production and diverse stress responses, including thermotolerance and antifungal drug resistance, of C. neoformans. This study provides further insights into the signaling network of CA/CO2-sensing pathway in pathogenic fungi.

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