Project description:To identify possible molecular mechanisms with a role in root branching initiation in S. moellendorffii, we employed the developmental root branching assay for an RNA-seq experiment and sampled root tips on each day from 0 to 5 d after the first branching (time point 0), in which 300 µm apical parts were sampled to enrich for the meristematic region, while non-meristematic root regions were sampled separately
Project description:E5.0 entails an intermediate state for primitive endoderm (PE) to visceral endoderm (VE) differentiation: a process required for proper embryonic development. Hnf1b is essential for primitive to visceral endoderm differentiation from E4.5 to E5.5. To understand development at E5.0 and pinpoint the role Hnf1b in proper embryonic development, embryos were harvest at E5.0 and single cells from three WT embryos and 3 Hnf1b KO embryos were isolated, dissociated, and scRNA sequenced using smartseq2. For experiments on WT embryos, CD1 females were crossed with CD1 or homozygous E-cadherin-GFP males. Hnf1b -/+22 line was derived through IVF of oocytes of Black6 mice with EMMA strain EM:07827(B6.129P2(D2)-Hnf1btm1Sce/Orl) Sperm, that was bought from the EMMA institution in Orleans.
Project description:This work aimed to characterize the molecular adaptations occurring in cork oak (Quercus suber) stems in adaptation to drought, and identify key genetic pathways regulating phellem development. One-year-old cork oak plants were grown for additional 6 months under well-watered (WW) or water-deficit (WD) conditions and main stems were targeted for transcriptomic analysis. WD had a negative impact on secondary growth, decreasing the activity of the vascular cambium and phellogen. Following a tissue-specific approach, we analyzed the transcriptional changes imposed by WD in phellem (outer bark), inner bark, and xylem, and found a global downregulation of genes related to cell division, cell wall biogenesis, lignin and/or suberin biosynthesis. Phellem and phloem showed a concerted upregulation of photosynthesis-related genes, suggesting a determinant role of stem photosynthesis in the adaptation of young plants to long-term drought. The data gathered will be important to further harness the diverse genetic background of this species for the development of optimized management practices.
Project description:This study describes the transcriptome of distinct blood compartments derived from whole blood collected from piglets. The dataset includes RNA-seq data from 20 samples representing five blood compartments: whole blood, serum, plasma, serum-derived exosomes, and plasma-derived exosomes. From each whole blood sample, the different fractions were carefully prepared using optimized and compartment-specific protocols. RNA-seq libraries were prepared using the LEXOGEN QuantSeq 3′ mRNA-Seq V2 Library Prep Kit with protocol adaptations for whole blood, plasma, serum, and exosome samples. Sequencing was performed on the Illumina NextSeq2000 platform in SR75. The 20 raw demultiplexed FASTQ files are part of this accession. Bioinformatic pre-processing was conducted using the Nextflow nf-core/rnaseq pipeline to ensure high sequence quality and robust transcriptome assembly and quantification. Each file contains raw sequencing reads along with base quality scores. Bioinformatic and statistical analyses explored the variability in gene detection and gene expression variability according to the five blood compartments.
Project description:TurboID-based proximity labeling of DMR6 and DLO1 identified several candidate interactors, including a putative F-box protein that may contribute to their SCF-dependent turnover.
Project description:Transcriptome analyses of the fission yeast Schizosaccharomyces pombe cells lacking the CSL transcription factor cbf11. Wild-type cells were used as a control. Both genotypes were analyzed when grown to exponential phase in the complex YES medium, or in YES supplemented with ammonium chloride. The aim was to identify Cbf11-regulated genes, and cbf11-associated phenotypes are affected by the presence of a good nitrogen source.
Project description:Proteomic investigation of Arabidopsis thaliana ftsH12 - ftsH12 is one of 17 genes of the FtsH metallo-protease family encoded within the A. thaliana genome.
Project description:Genome-wide identification of transcription factor (TF) binding sites in the genome of the fission yeast Schizosaccharomyces pombe. The ChIP-nexus method was used. TFs included were: Cbf11-TAP and Cbf12-TAP (and their DBM mutants with impaired DNA binding), TAP-Mga2, and Fkh2-TAP (as an irrelevant control TF). IPs from an untagged WT strain were also analyzed. Cbf11-related IPs were performed from exponential cultures, while Cbf12-related IPs were performed from stationary cultures. YES complex medium was used for all cultivations.
Project description:Protein oxidation results from the reaction of amino-acid side chains with reactive oxygen species (ROS) and is mostly irreversible. In non-photosynthetic tissues, mitochondria are a main source of ROS, whereas plastids are the major source in photosynthetic tissues. Oxidized proteins suffer from decreased structural integrity and even loss of function, and their accumulation ultimately leads to cytotoxic aggregates. In mammals, this correlates with aging and is linked to several age-related pathologies. Mammalian proteolytic pathways for clearance of oxidized proteins are under intensive research, while mechanistic insights into this process in plants is scarce. AARE enzymes are ATP-independent serine proteases presumably acting on oxidized proteins and operating in a dual exo-/endopeptidase mode. They are found in all domains of life. Here, we investigated AARE enzymes in the moss Physcomitrella and the angiosperm Arabidopsis and identified three homologous nuclear genes in Physcomitrella (PpAARE1-3) and a single nuclear gene in Arabidopsis (AtAARE). Surprisingly, we observed triple localization of the proteins AtAARE and PpAARE1 to plastids, mitochondria and the cytosol in vivo, likely conserved across the plant lineage. This represents an ATP-independent possibility for degradation of oxidized proteins in the major source organelles of ROS in plants, which is distinct to mammals. Combinatorial knockout plants and protein interaction analysis revealed specific interactions of the moss AARE isoforms and functions in progressive aging. Analysis of an AtAARE T-DNA mutant further suggests conservation of AARE function in age-related development.
Project description:Nucleic acid and histone modifications critically depend on central metabolism for substrates and co-factors. Although a few enzymes related to the formation of these required metabolites have been reported in the nucleus, the corresponding metabolic pathways are considered to function elsewhere in the cell. Here we show that a substantial part of the mitochondrial tricarboxylic acid (TCA) cycle, the biosynthetic hub of epigenetic modification factors, is operational also in the nucleus. Using 13C-tracer analysis, we identified activity of glutamine-to-fumarate, citrate-to-succinate, and glutamine-to-aspartate routes in the nuclei of HeLa cells. Proximity labeling mass-spectrometry revealed a spatial vicinity of the involved enzymes with core nuclear proteins, supporting their nuclear location. We further show nuclear localization of aconitase 2 and 2-oxoglutarate dehydrogenase in mouse embryonic stem cells. Together, our results demonstrate operation of an extended metabolic pathway in the nucleus warranting a revision of the canonical view on metabolic compartmentalization and gene expression regulation. This dataset includes the data relating to the use of LOPIT to determine the subcellular localisation of succinylated proteins.