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Coccidioides posadasii strain:Silveira

ABSTRACT: Improved high-quality genome assembly and annotation of Coccidioides posadasii, Silveira strain

PROVIDER: PRJNA494320 | ENA |

REPOSITORIES: ENA

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			Action	DRS
		Other
	SRR9644374_1.fastq.gz	Fastqsanger.gz
	SRR9644374_2.fastq.gz	Fastqsanger.gz
	SRR9644375_subreads.fastq.gz	Fastqsanger.gz

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Proteogenomic Re-annotation of Coccidioides posadasii strain Silveira

Project description:The aims of this study were to present modifications to the annotations of the genome of C. posadasii, one of two closely related species of Coccidioides, a dimorphic fungal pathogen that causes coccidioidomycosis, also called Valley Fever. Proteins present in lysates and filtrates of in vitro grown mycelia and parasitic phase spherules from C. posadasii strain Silveira were analyzed using a GeLC-MS/MS method.

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AÂ Coccidioides posadasii CPS1Â deletion mutant is avirulent and protects mice from lethal infection

Project description:TheÂ CPS1Â gene was identified as a virulence factor in the maize pathogen, Cochliobolus heterostrophus. Hypothesizing that the homologous gene in Coccidioides posadasiiÂ (Cp) could be important for virulence, we created a deletion mutant,Â Îcps1, which was unable to cause disease in three strains of mice (C57BL/6, BALB/c, or the severely immunodeficient NOD-scid,Î³cnullÂ [NSG]). Only a single colony was recovered from one of 60 C57BL/6 mice following intranasal infections of up to 4400 spores. Following administration of very high doses (10,000 to 2.5 x 10^7Â spores) to NSG and BALB/c mice, spherules were observed in lung sections at time points from day 3 to day 10 post-infection, but nearly all appeared degraded with infrequent endosporulation. Although the role ofÂ CPS1Â in virulence is not understood, phenotypic alterations and transcription differences of at least 33 genes inÂ Îcps1Â vs. Cp is consistent with both metabolic and regulatory functions for the gene. The inÂ vitroÂ phenotype ofÂ Îcps1Â showed slower growth of mycelia with delayed and lower spore production compared to Cp, andÂ in vitro spherules were smaller. Vaccination of C57BL/6 or BALB/c mice with liveÂ Îcps1 spores either intranasally, intraperitoneally or subcutaneously resulted in over 95% survival with mean residual lung fungal burdens <1000 colony-forming units from an otherwise lethal Cp intranasal infection. Considering its apparently complete attenuation of virulence and the high degree of resistance to Cp infection when used as a vaccine,Â Îcps1Â is a promising vaccine candidate for preventing coccidioidomycosis in humans or other animals. Wild type and CPS1 deletion mutant strains of Coccidioides posadasii strain Silveira spherules grown for 48 hours in Converse medium at 38 degrees celsius in duplicate. RNAseq was performed on an Illumina HiSeq2000 (2x100 paired end).

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The WOPR family protein Ryp1 is a key regulator of gene expression, development, and virulence in the thermally dimorphic fungal pathogen Coccidioides posadasii

Project description:Coccidioides spp. are fungal pathogens endemic to Southern California, Arizona, and other desert regions where they cause a spectrum of symptoms known as Valley Fever in otherwise healthy individuals. In the soil, Coccidioides grows in a hyphal form that produces vegetative spores (arthroconidia). When arthroconidia are inhaled by mammals they undergo an elaborate developmental transition consisting of germination, multiple rounds of nuclear division, and segmentation to form large spherules filled with vegetative endospores. Very little is understood about the molecular basis of spherule formation. Here we characterize the role of the conserved transcription factor Ryp1 in Coccidioides spherule formation. We show that Coccidioides ryp1 mutants are unable to form mature spherules, instead arresting at an early stage of spherulation. We analyze the transcriptional profile of wild-type and Δryp1 mutant cells under hyphal and spherule-promoting conditions, thereby defining a set of hyphal- or spherule-enriched transcripts that are dependent on Ryp1 for their expression. Ryp1-dependent transcripts include key virulence factors such as SOWgp, which encodes the spherule outer wall glycoprotein. In the mouse model of coccidioidomycosis, we find that the Δryp1 mutant is completely avirulent, indicating that Ryp1-dependent pathways are essential for the ability of Coccidioides to cause disease. Vaccination of C57BL/6 mice with live Δryp1 spores does not provide any protection from lethal C. posadasii intranasal infection, consistent with our findings that the Δryp1 mutant fails to make mature spherules. Taken together, this work identifies the first transcriptional regulator that drives mature spherulation and virulence in Coccidioides.

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