Project description:Background: Microorganisms are the major cause of food spoilage during storage, processing and distribution. Pseudomonas fluorescens is a typical spoilage bacterium that contributes to a large extent to the spoilage process of proteinaceous food. RpoS is considered an important global regulator involved in stress survival and virulence in many pathogens. Our previous work revealed that RpoS contributed to the spoilage activities of P. fluorescens by regulating resistance to different stress conditions, extracellular acylated homoserine lactone (AHL) levels, extracellular protease and total volatile basic nitrogen (TVB-N) production. However, RpoS-dependent genes in P. fluorescens remained undefined. Results: RNA-seq transcriptomics analysis combined with quantitative proteomics analysis basing on multiplexed isobaric tandem mass tag (TMT) labeling was performed for the P. fluorescens wild-type strain UK4 and its derivative carrying a rpoS mutation. A total of 375 differentially expressed genes (DEGs) and 212 differentially expressed proteins (DEPs) were identified in these two backgrounds. The DGEs were further verified by qRT-PCR tests, and the genes directly regulated by RpoS were confirmed by 5’-RACE-PCR sequencing. The combining transcriptome and proteome analysis revealed a role of this regulator in several cellular processes, including polysaccharide metabolism, intracellular secretion and extracellular structures, cell well biogenesis, stress responses, ammonia and biogenic amine production, which may contribute to biofilm formation, stress resistance and spoilage activities of P. fluorescens. Moreover, in this work we indeed observed that RpoS contributed to the production of the macrocolony biofilm’s matrix.
Project description:In a previous study, we found that H2S alleviates salinity stress in cucumber by maintaining the Na+/K+ balance and by regulating H2S metabolism and the oxidative stress response. However, little is known about the molecular mechanisms behind H2S-regulated salt-stress tolerance in cucumber. Here, an integrated transcriptomic and proteomic analysis based on RNA-seq and 2-DE was used to investigate the global mechanism underlying H2S-regulated salt-stress tolerance. In total, 11 761 differentially expressed genes (DEGs) and 61 differentially expressed proteins (DEPs) were identified. Analysis of the pathways associated with the DEGs showed that salt stress enriched expression of genes in primary and energy metabolism, such as photosynthesis, carbon metabolism and biosynthesis of amino acids. Application of H2S significantly decreased these DEGs but enriched DEGs related to plant-pathogen interaction, sulfur-containing metabolism, cell defense and signal transduction pathways. Notably, changes related to sulfur-containing metabolism and cell defense were also observed through proteome analysis, such as Cysteine synthase 1, Glutathione S-transferase U25-like, Protein disulfide-isomerase and Peroxidase 2. We present the first global analysis of the mechanism underlying H2S regulation of salt-stress tolerance in cucumber through tracking changes in the expression of specific proteins and genes.
Project description:The two subunits of IL-35, EBI3 and P35, were fused together and transfected into Panc-1 cell via lentivirus. The sequence of the fused gene is identical to that of a commercial IL-35-overexpessed plasmid (InvivoGEN, pORF9-hIL35elasti). An empty vector was used as the control. The two cell lines were subjected to a genome-wide RNA sequencing.
Project description:To screen out the downstream genes of IL-35 The two subunits of IL-35, EBI3 and P35, were fused together and transfected into Panc-1 cell via lentivirus. The sequence of the fused gene is identical to that of a commercial IL-35-overexpessed plasmid (InvivoGEN, pORF9-hIL35elasti). An empty vector was used as the control. The two cell lines were subjected to a genome-wide RNA sequencing.
Project description:Introduction: Post-traumatic stress disorder (PTSD) is highly prevalent among U.S. service members and veterans (SMV) and has lasting impacts on health and well-being. However, the biological underpinnings of PTSD remain poorly characterized. This study aimed to discover novel candidate proteins and protein pathways associated with PTSD using unbiased, high-throughput proteomics profiling. Methods: A cross-sectional study was conducted using a subset of SMV participants from a clinical cohort undergoing evaluations at the National Intrepid Center of Excellence, Walter Reed National Military Medical Center, who consented to a research blood draw and use of their clinical data in research. The cohort with available blood samples was classified into PTSD-Present and PTSD-Absent groups based on the PTSD Checklist- Civilian Version. Olink high-throughput proteomic profiling examined 5400 human plasma proteins, and differentially expressed proteins were examined. Ingenuity pathway analysis was used to identify proteomic pathways among the significant differentially expressed proteins between the PTSD-Present and PTSD-Absent groups. Results: We included 208 samples in our analysis, 126 with and 82 without PTSD. We identified 366 proteins that were significantly differentially expressed between groups, with the 3 most significant being LMOD2 (Leiomodin- 2), ATP5F1D (ATP synthase δ-subunit), and CASKIN1 (CASK-interacting protein). Extracellular matrix organization pathways and Vascular Endothelial Growth Factor (VEGF) signaling were downregulated in PTSD, suggesting possible vascular inflammation and remodeling. Data- driven protein networks suggest reduced immune cell activation. Discussion: Determining differential protein expression and identifying the associated protein pathways linked to PTSD may provide new insights into the biological basis of chronic PTSD symptoms and help identify novel candidate protein biomarkers of PTSD and PTSD symptoms for validation in separate and larger cohorts. From this discovery cohort, we report that elevated PTSD symptoms may be associated with downregulation of extracellular matrix organization, inflammation signaling, and vascular remodeling pathways. Future research is necessary to validate this novel group of biomarkers and pathways.
Project description:Data on absolute molecule numbers are rare but will empower the modeling, understanding and comparison of cellular functions and biological systems. We quantified transcriptomes and proteomes in fission yeast during both proliferation and quiescence. This rich resource provides the first comprehensive reference for all RNA and most protein concentrations in a eukaryote under key physiological conditions. This integrated dataset will support quantitative biology and afford unique biological insights into cell regulation. While mRNAs are typically expressed in a narrow range above 1 copy/cell, most long non-coding RNAs, except for a specific subset, are strongly repressed below 1 copy/cell. Cell cycle-regulated transcription tunes mRNA numbers to phase-specific requirements but can also lead to switch-like expression. Proteins greatly exceed mRNAs in abundance and dynamic range, and their numbers scales with functional demands. Upon transition to quiescence, the proteome composition changes substantially but, in stark contrast to mRNAs, proteins do not uniformly decrease but scale with cell volume.
Project description:The aim of this experiment was to identify transcripts upregulated during vernalization in Lolium perenne plants. Illumina Genome Analyzer II RNA-Seq data was generated from leaf samples collected before vernalization, and after the start of cold treatment, at 2 days, 4 weeks and 9 weeks.
Project description:Over 20% of Earth’s terrestrial surface is underlain by permafrost that represents one of the largest terrestrial carbon pools, with an estimated ~1700 Pg of carbon (C) contained in the upper 3 m of permafrost. Models estimate that C release from thawing permafrost might represent the largest new transfer of C from the biosphere to the atmosphere as the climate warms. Here we investigated microbial community phylogeny, genetic functional potential gene expression, and protein production patterns along a natural thaw gradient, including permafrost, the seasonally thawed active layer and nearby thawed thermokarst bog, using a combination of molecular “omics” approaches: metagenomics (MG), metatranscriptomics (MT) and metaproteomics (MP). Highlights from these analyses reveal energy yielding microbial processes and potential strategies for microbial survival in permafrost soils, and linkages between biogeochemical process rates and –omics measurements. The results provide new knowledge about microbial life and activity potential in permafrost, the potential importance of iron reduction as a survival strategy under frozen conditions in mineral soils, and the importance of methanogenesis following thaw. The multi-omics strategy demonstrated here enables better mechanistic understanding of the ecological strategies utilized by soil microbial communities in response to climate change. Associated metagenomics data available at the EBI Metagenomics portal under the accession number <a href="https://www.ebi.ac.uk/metagenomics/projects/SRP052575">SRP052575</a>.
Project description:This study addresses this gap by conducting a direct comparison of eight platforms, representing both affinity-based and diverse mass spectrometry approaches, and covering over 13,000 proteins. By applying these platforms to the same cohort, we systematically assess their performance, identifying key differences and complementary strengths. Our findings offer valuable insights for researchers, highlighting trade-offs in coverage and their implications for biomarker discovery and clinical applications. This study serves as an essential resource, offering both technical evaluation and biological insights to support the development of novel diagnostics and therapeutics through plasma proteomics.
Project description:This study addresses this gap by conducting a direct comparison of eight platforms, representing both affinity-based and diverse mass spectrometry approaches, and covering over 13,000 proteins. By applying these platforms to the same cohort, we systematically assess their performance, identifying key differences and complementary strengths. Our findings offer valuable insights for researchers, highlighting trade-offs in coverage and their implications for biomarker discovery and clinical applications. This study serves as an essential resource, offering both technical evaluation and biological insights to support the development of novel diagnostics and therapeutics through plasma proteomics.