Project description:We used Candida albicans lab strain SC5314 to obtain tunicamycin adaptors. We did whole genome sequencing of the adaptors and the parent as well.
Project description:We exposed Candida parapsilosis clinical isolate #12108 to YPD plate supplemented with 8µg/ml of tunicamycin. We randomly selected 18 adaptors. We did sequencing of these adaptors.
Project description:Candida auris clade III isolate B11221 was spread on YPD plate supplemented with 8 µg/ml tunicamycin. Randomly 18 adaptors were chosen for further analysis. We did sequencing of these 18 adaptors as well as the parent.
Project description:Candida auris clade III isolate B12039 was spread on YPD plate supplemented with 8 µg/ml tunicamycin. Randomly 27 adaptors were chosen for further analysis. We did sequencing of these 27 adaptors as well as the parent.
Project description:WalKR is a two-component system that is essential for viability in Gram-positive bacteria that regulates the all-important autolysins. Here we show a T101M mutation in walR confers a defect in autolysis, a thickened cell wall and decreased sensitivity to antibiotics that target the lipid II cycle, a phenotype that is reminiscent of the clinical resistance form known as vancomycin intermediate-resistant Staphylococcus aureus. Importantly, this is accompanied by dramatic sensitization to tunicamycin. We demonstrate that this phenotype is due to partial collapse of a pathway consisting of autolysins, AtlA and Sle1, a transmembrane sugar permease, MurP, and GlcNAc recycling enzymes, MupG and MurQ. We suggest that this causes a shortage of substrate for MraY causing it to be hypersensitive to competitive inhibition by tunicamycin. This constitutes a new molecular model for antibiotic sensitivity in S. aureus and a promising new route for antibiotic discovery.
Project description:Polysomes of untreated (NT) or tunicamycin-treated (TM) human cardiomyocyte AC16 cells were immunoprecipitated (IP MRPS15) with anti-MRPS15 antibody, followed by RNA sequencing. As a control, RNAseq was performed on polysome-associated RNAs of untreated or tunicamycin-treated human cardiomyocyte AC16 cells before immunoprecipitation (input).
Project description:Purpose: Evaluate transcriptional changes in glioma derived stem cells, U251, and normal human astrocytes following treatment with NH125, Tunicamycin, or 0.1% DMSO Methods: Glioma derived stem cells, U251 and Normal Human Astrocytes were plated in individual dishes and treated with either 2.5 micromolar NH125, 1.0 mcg/mL Tunicamycin, or 0.1% DMSO (vehicle) for twenty-four hours followed by total RNA extraction, library preparation and next generation sequencing Results: Using our analysis pipeline we mapped our sequence reads to the reference genome GRCh38. Following read count normalization and differential expression of NH125 and tunicamycin treated samples to vehicle treated controls we found that NH125 and tunicamycin treatment lead to activation of similar signalling pathways Conclusions: We present an optimized workflow for analysis of transcriptional changes in drug treated glioma derived stem cells, glioblastoma cells and normal human astrocytes
Project description:RNA sequencing on LNCaP cells was carried out to study how tunicamycin-induced gene expression is affected by knockdown of EIF2AK3 and ATF4.
Project description:Approximately 1 million cells of Candida glabrata lab strian BG2 were spread on YPD plate supplemented with 8 ug/ml tunicamycin. Randomly 30 adaptors were chosen. 28 adpators were more tolerant than parent to tunicamycin. These 28 tolerant adaptors, as well as the parent BG2 were sequenced.
Project description:To further development of our miRNA expression approach to ER stress, we have employed miRNA microarray expression profiling as a discovery platform to identify ER stress-responsible ones. Mouse embryonic fibroblasts (MEFs) deficient in a ER stress mediator XBP1 were treated with tunicamycin for 12 or 24 hrs. miRNAs responsible for tunicamycin-treatment for 12hrs in XBP1-dependent manner were extracted. Among them, expression of three miRNAs (miR-23a, miR-27a, miR-24-2) was quantified in the RNA samples from the same as the microarray, and COS7 cells by real-time PCR, confirming existence of similar mechanisms of trancriptional repression in ER stress by tunicamycin treatment.
