Ontology highlight
ABSTRACT:
PROVIDER: PRJNA530034 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR8837390_1.fastq.gz | Fastqsanger.gz | |||
| SRR8837390_2.fastq.gz | Fastqsanger.gz | |||
| SRR8837410_1.fastq.gz | Fastqsanger.gz | |||
| SRR8837410_2.fastq.gz | Fastqsanger.gz | |||
| SRR8837391_1.fastq.gz | Fastqsanger.gz |
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Project description:Isolates of Cryptococcus neoformans, a fungal pathogen that kills almost 200,000 people worldwide each year, differ at a few thousand up to almost a million DNA sequence positions compared to a 19-megabase reference genome. We used bulked segregant analysis and association analysis, genetic methods that require no prior knowledge of sequence function, to address the key question of which naturally occurring variants influence fungal virulence. We identified a region containing such variants, prioritized them, and engineered strains to test our findings in a mouse model of infection. At one locus we identified a 4-nt variant in the PDE2 gene, which severely truncates its phosphodiesterase product and significantly alters virulence. Our studies demonstrate a powerful and unbiased strategy for identifying key genomic regions in the absence of prior information, suggest revisions to our assumptions about cAMP levels and about common laboratory strains, and provide significant sequence and strain resources to the community.
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Project description:For a eukaryotic genome to properly function, its chromatin must be precisely organized, as genome topology impacts transcriptional regulation, cell division, DNA replication, and DNA repair, among other essential processes. Disruption of human genome topology can lead to disease states, such as cancer. The advent of chromosome conformation capture with high-throughput sequencing (Hi-C) technologies to assess genome organization has revolutionized our understanding of the arrangement of chromosomes within the nuclear genome. Critical developments include chromosomal territories, active/silent chromatin compartments, Topologically Associated Domains (TADs), and chromatin loops. However, to fully elucidate folding principles at the gene level, Hi-C datasets at an extremely high resolution are required: while low resolution (e.g., 40 kb bin) heatmaps can provide important insights into chromosomal level contacts, lower resolution datasets cannot provide information about individual gene or promoter contacts. Thus, high-resolution Hi-C datasets can elucidate principles of folding, including those of model organisms whose chromosome conformation can elucidate the folding of the human genome. Here, we have examined the high-resolution genome topology of the model fungal organism Neurospora crassa: our composite Hi-C dataset, generated using two restriction enzymes to monitor euchromatin (DpnII) and heterochromatin (MseI), provides exquisite detail for both larger chromosomal structures and individual gene contacts, including how repressed euchromatic genes associate with constitutive heterochromatic regions. Our high-resolution Neurospora Hi-C datasets should be an outstanding resource to the fungal community and provide valuable insights to the folding of higher organism genomes.
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Project description:Purpose: Breeding for gibberella ear rot resistance have been challenging due to the high complexity of the trait. The current study attempts to characterize defence responses to Fusarium graminearum infection in two maize inbred lines with different levels of resistance to the pathogen. Methods: RNA was extracted from developing kernels of two inbred lines, which had been either fungal (F. graminearum DAOM180378) or mock inoculated 11 days post sibcrossing, using a guanidine isothiocyanate method and ultra-centrifugation with cesium chloride. Isolated RNA was used to quantify whole genome gene expression using RNA-seq (Illumina TruSeq RNA library prep kit v2, Illumina HiSeq 2000). Paired end reads generated from RNA-seq were trimmed of adaptors and low quality reads, aligned with the B73 reference genome sequence version 2, expression levels (TPM) were computed and differential gene expression analysis were performed using CLC Genomics Workbench version 9. Results: Gene transcripts responding to fungal infection were captured by comparing gene expression levels in mock and fungal inoculated maize ears and gene ontology terms associated with significantly up-regulated gene transcripts were determined for each inbred. More genes were up regulated in the susceptible inbred relative to the resistant inbred, many of which are associated with oxidation-reduction processes potentially causing earlier programmed cell death in the susceptible inbred. Conclusions: This information helped to identify gene transcripts that were relevant in defense responses with potential applicability in routine breeding efforts and to propose an effective GER resistance mechanism.
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