Identification of NTRC interacting partners during cold acclimation
Ontology highlight
ABSTRACT: We performed co-immunoprecipitation experiments to identify interacting proteins of NADPH-dependent thioredoxin reductase C (NTRC) at 15 °C and 23 °C.
Project description:In this study transcriptomic data of three life history stages of Orciraptor agilis was generated: 1) Gliding cells in absence of food ('gliding'), 2) Cells attached to the cell wall of its algal prey during perforation ('fattacking'), 3) Cells after acquisition of the algal plastid material ('digesting'). Furthermore, RNA-seq of the algal prey Mougeotia sp. was also performed. A de novo transcriptome assembly of the algal reads was performed in order to identify and substract algal reads of the Orciraptor samples by mapping the Orciraptor reads to the algal transcriptome. After this filtering step the remaining Orciraptor reads from all libraries were pooled for a de novo transcriptome assembly of Orciraptor agilis. This transcriptome was the basis for a comparative transcriptomic study in which transcript expression was compared between the three life history stages.
Project description:One of the components of the tumor progression of melanoma of the skin is the formation of drug resistance. Chemotherapeutic agents can affect the cell cycle, DNA repair. By influencing melanoma cells of the BRO and SK-MEL-2 lines, the effect of a cytostatic drug on the genomic profile was determined
Project description:On day 7 after B16 melanoma cells transplantation, C57Bl6 mice with subcutaneous palpable tumors were randomly divided into 2 groups for experimental treatment with Dacarbazine. Group 1 «Control (PBS) », n=5, animals in this group were intraperitoneally injected with phosphate buffered saline in a volume of 250 µl. Group 2 «DTIC», n=5, animals in this group were intraperitoneally injected with a solution of Dacarbazine (DTIC) (Sigma-Aldrich, USA) in phosphate buffer at a concentration of 50 mg/kg of animal weight. Injections with PBS or DTIC were carried out three times on days 8, 10 and 12 after melanoma cell transplantation, on day 14, mice were sacrificed for further analysis of primary tumors and organs.
Project description:Here we demonstrate by experimental evolution and genetics the rapid and repeatable evolutionary re-wiring of regulatory pathways, where the cellular nitrogen regulatory system acquired the ability to “cross-talk” with the flagellar regulon. This rewiring restored flagella to aflagellate strains of Pseudomonas fluorescens devoid of the master regulator fleQ via a repeatable two-step evolutionary pathway. Step 1 initiates cross-talk by increasing intracellular levels of phosphorylated NtrC , a distant homologue of FleQ, which begins to commandeer control of the fleQ regulon whilst constitutively activating nitrogen uptake and assimilation genes. Step 2 is a switch-of-function mutation (NtrC’) that further redirects NtrC towards activation of motility and away from nitrogen uptake. Evolved NtrC’ emerges with a novel function as a flagellar regulator. Our results demonstrate that natural selection can rapidly rewire regulatory networks in very few, repeatable mutational steps to adopt new functionality.
Project description:YAP transcriptional regulator controls cell mechanics by activating genes involved in cell-matrix interaction following extracellular matrix (ECM) remodelling and stiffening. YAP is needed for cardiogenesis in mouse but is repressed in adult cardiomyocytes. The protein is reactivated following ischemic insults, although the timing and mechanisms underlying YAP depletion during heart development and the reason for its reactivation are unclear. Here, we combine pluripotent stem cell (PSC) cardiac differentiation, mouse embryo development and human heart tissue analysis to demonstrate that the fine-tuning of cell mechanics, as controlled by YAP multiphasic activation through TEAD transcription, is crucial for mesoderm commitment and cardiac progenitor specification. Finally, by adopting induced PSC models of dilated cardiomyopathy, we prove that YAP-TEAD reactivation in diseased cardiomyocytes empowers calcium handling apparatus and increases cell contractility. Given YAP prompt activation following myocardial infarction, we unveil a novel role for mechanosensing in connecting ECM remodelling to cardiomyocyte function in pathological heart.
Project description:Quiescent canine valvular interstitial cells (n=3) and activated valvular interstitial cells (n=3) were grown in 2% FBS. Quiescent cells were treated with either TGFB1 or vehicle for 3 days and activated cells were treated with SB431542 for 4 days. mRNA was extracted using the qiagen RNeasy minikit and assessed by the agilent 2200 bioanalsyser. all samples passed quality control and transcriptomic analysis was performed by edinburgh genomics using the Affymetrix GeneChip™ Canine Gene 1.1ST Array. Results were reported as CEL files and then analysed in Affymetrix expression console (version 1.4.1.46) using the canine 1.1ST library files from Affymetrix. Robust Multi-array Average (RMA) was used to perform gene level normalisation and signal level summarisation.Following this, quality control assessment was performed on the data set. This included assessment of hybridisation (spike in) controls, labelling (poly-A) controls and area under the curve (AUC) control, probe cell intensity boxplots, a signal histogram graph and principle component analysis (PCA) plot. Following this assessment, the datasets were exported with annotation as a text file or as a CHT file. Affymetrix transcriptome analysis console (version 3.1.0.5) was used to perform paired or unpaired one-way analysis of variance (ANOVA).
Project description:Arabidopsis thaliana wildtype (Col-0) was compared with the ntrc mutant under different photoperiods (short day: 8h/16h; long day 16h/8h light/dark) and different ages (10d and 28/21d).
Project description:We performed chromatin run on and sequencing (ChRO-seq) on TeloHAEC cells before and after TNFα stimulation to map locations of RNA polymerase and quantify nascent transcription at RELA peaks.
Project description:We show that NtrC couples the Ntr stress response and stringent response in N starved E. coli, which appears to be a conserved adaptive strategy employed by many bacteria to manage conditions of nutritional adversity. N starved Escherichia coli initiate the nitrogen regulation (Ntr) stress response as an adaptive mechanism to scavenge for alternative N sources. The Ntr stress response requires the global transcriptional regulator nitrogen regulatory protein C (NtrC). We discovered that the transcription of relA, the key gene responsible for the synthesis of the major effector nucleotide alamorne of the bacterial stringent response, guanosine pentaphosphate (ppGpp), is positively regulated by NtrC in N starved E. coli. we addressed Ntr stress response-ppGpp alarmone links and mapped the genome-wide binding targets of NtrC in E. coli during N starvation using chromatin immunoprecipitation (ChIP) followed by high-throughput sequencing (ChIP-seq) to gain insight into the NtrC-dependent gene networks. To identify candidate genome regions that are preferentially associated with NtrC, we introduced an in-frame fusion encoding three repeats of the FLAG epitope to the 3-prime end of glnG in E. coli strain NCM3722, a prototrophic E. coli K-12 strain.
Project description:DNA Double Strands Breaks (DSBs) are highly detrimental since they can lead to mutations and chromosomes rearrangements (amplification, deletion, translocation and chromosome loss). Here, we set to assess the role of senataxin, a RNA:DNA helicase involved in the regulation of transcription and the maintenance of genome integrity, at sites of DNA Double Strand Breaks. We performed ChIP-Seq mapping of senataxin before and after damage, genome-wide DNA:RNA hybrids (DRIP) mapping before and after damage. We also performed RNA-Seq and RNA pol II mapping (total and phosphorylated on serine 2 of the CTD) by ChIP-Seq in undamaged cells to discriminate between damage in active versus inactive regions.