Ontology highlight
ABSTRACT:
PROVIDER: PRJNA772181 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR16464239.fastq.gz | Fastqsanger.gz | |||
| SRR16464240.fastq.gz | Fastqsanger.gz | |||
| SRR16464241.fastq.gz | Fastqsanger.gz | |||
| SRR16464242.fastq.gz | Fastqsanger.gz | |||
| SRR16464243.fastq.gz | Fastqsanger.gz |
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Project description:TnSeq mapping and BarSeq quantification of Pectobacterium mutant fitness
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Project description:Transposon insertion site sequencing (TIS) is a powerful tool that has significantly advanced our knowledge of functional genomics. While providing valuable insights, these applications of TIS focus on (conditional) gene essentiality and neglect possibly interesting but subtle differences in the importance of genes for fitness. Notably, data from TIS experiments can be used for fitness quantification and constructing genetic interaction maps, though this potential is only sporadically exploited. We aimed to develop a method to quantify the fitness of gene disruption mutants using data obtained from the TIS screen SATAY. This dataset was used to determine the reproducibility of the fitness estimates across biological and technical replicates of the same strain of S. cerevisiae. In addition, a mutant bem3∆ strain was utilized to compare the genetic interactions inferred from these fitness estimates with those documented in published databases. The dataset for the wild-type strain was created by transforming strain yWT01 with plasmid pBK549 and picking 4 different colonies from the transformation plate. These 4 biological replicates were then renamed to FD7, FD9, FD11 and FD12.
2024-11-18 | E-MTAB-14476 | biostudies-arrayexpress
Project description:A systematic approach allowing the identification of the molecular way-of-action of novel potential drugs represents the golden-tool for drug-discovery. While high-throughput screening technologies of large libraries is now well established, the assessment of the drug targets and mechanism of action is still under development. Taking advantage of the yeast model Saccharomyces cerevisiae, we herein applied BarSeq, a Next Generation Sequencing-based method to the analysis of both haploinsufficiency and homozygous fitness effects of a novel antifungal drug ('089') compared to the well-known antifungal ketoconazole. '089' was a novel compound identified in during a screen for antifungal drugs, as it was showing fungicidal effects, and able to affect the yeast fitness at the mitochondrial level (Stefanini et al., 2010. (Dissection of the Effects of Small Bicyclic Peptidomimetics on a Panel of Saccharomyces cerevisiae Mutants;.J Biol Chem, 285: 23477-23485.) Integrative bioinformatic analysis of BarSeq, whole genome expression analysis and classical biological assays identified the target and cell pathways affected by the novel antifungal. Confirmation of the effects observed in the yeast model and in pathogenic fungi further demonstrated the reliability of the multi-sided approach and the novelty of the targets and way-of-action of the new class of molecules studied representing a valuable source of novel antifungals.
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Project description:Lytic bacteriophages able to infect and kill Dickeya spp. can be readily isolated from virtually all Dickeya spp.-containing environments, yet little is known about the selective pressure those viruses exert on their hosts. Here, we identified two spontaneous phage-resistant D. solani IPO 2222 mutants, DsR34 and DsR207, resistant to infection caused by phage vB_Dsol_D5 (ΦD5) that expressed a reduced ability to macerate potato tuber tissues compared to the wild-type, phage-susceptible D. solani IPO 2222 strain. Genome sequencing revealed that mutants had point mutations in two genes encoding: secretion protein HlyD (mutant DsR34) and elongation factor Tu (EF-Tu) (mutant DsR207). Both mutations impacted the proteoms of D. solani grown in rich and minimal media. Furthermore, DsR34 and DsR207 were characterized for features essential for their ecological success in a plant environment, including the ability to use various carbon and nitrogen sources, production of plant cell wall degrading enzymes, ability to form biofilms, siderophore production, swimming and swarming motility and virulence in planta. Compared to the wild-type ΦD5-susceptible D. solani strain, mutants DsR34 and DsR207 expressed reduced ability to macerate chicory leaves and to colonize and cause symptoms in growing potato plants. The implications of the ΦD5 resistance on the ecological performance of D. solani are discussed.
2023-04-28 | PXD038825 | Pride
Project description:The DsbA oxidoreductase is a crucial factor responsible for introduction of disulfide bonds to the extracytoplasmic proteins in bacteria. A lack of the proper disulfides frequently leads to instability and/or loss of protein function. In pathogens, numerous envelope and extracellular proteins play important roles in pathogenesis; therefore, their improper disulfide bonding may lead to avirulent phenotypes. The importance of the DsbA function in phytopathogens has not been extensively studied yet. Dickeya solani is a bacterium from the Soft Rot Pectobacteriaceae group which is responsible for very high economic losses mainly on potato. In recent years, D. solani became the most abundant potato pathogen among Dickeya species in Europe. In this work, using the D. solani dsbA mutant, we demonstrated that a lack of the DsbA function caused loss of virulence. Mutant bacteria were deficient in most secreted virulence determinants and were not able to develop disease symptoms in the natural host, the potato plant. The SWATH-MS-based proteomic analysis revealed that the dbsA mutation led to multifaceted effects in the D. solani cells. First of all, the levels of the majority of plant cell wall degrading enzymes and proteins related to motility and chemotaxis were severely reduced. Furthermore, the protein profiles suggested induction of the envelope and cytoplasm stress responses in the mutant cells. Finally, the outer membrane barrier seemed to be disturbed by the mutation. Our results clearly demonstrate that the function played by the DsbA oxidoreductase is indispensable for D. solani virulence and a lack of DsbA significantly disturbs cellular physiology. A thorough analysis of proteomic research suggests that a lack of virulence may result from both, abnormalities of the disulfide deprived virulence determinants and the envelope stress-dependent repression of the virulence genes in the dsbA mutant.
2022-02-15 | PXD028047 | Pride