Project description:Homo sapiens fresh whole blood was infected with Candida parapsilosis. RNA-pool of both species extracted at 0min (control), 15, 30, 60, 120, 240 min. Samples are rRNA depleted. Measurement of Homo sapiens gene expression.
Project description:Homo sapiens fresh whole blood was infected with Candida albicans SC5314. RNA-pool of both species extracted at 0min (control), 15, 30, 60, 120, 240 min. Samples are rRNA depleted. Expression measurement of Homo sapiens genes.
Project description:E. coli CyDisCo strain enables a high yield secretion of disulfide bond-containing proteins to the periplasm via Twin-arginine (Tat) pathway. Introducing two exogenous oxidases: the yeast sulfydryl oxidase (Erv1p) and human protein disulfide isomerase (PDI), the CyDisCo strain changes the cytoplasm into an oxidized environment, where the disulfide bonds can efficiently be formed. In this study, we analyzed the proteome changes upon the expression of disulfide bond-containing scFv and the misfolded scFv in the CyDisCo strain. The correctly folded protein is secreted to the periplasm, while the misfolded protein accumulates exclusively in the inclusion body fraction. We observed a high number of significant changes mostly in proteins associated with protein folding and degradation, oxidative stress, membrane transport and integrity.
Project description:Thiol-dependent redox regulation is essential for the rapid adaptation of chloroplast metabolism to unpredictable changes of light intensity. The disulfide reductase activity of thioredoxins (Trxs), which relies on photo-reduced ferredoxin (Fdx) and a Fdx-dependent Trx reductase (FTR), constitutes the Fdx-FTR-Trxs system, which links chloroplast redox regulation to light. In addition, chloroplasts harbor an NADPH-dependent Trx reductase (NTR) with a joint Trx domain, NTRC. The activity of these two redox systems is integrated by the balance of the hydrogen peroxide scavenging enzyme 2-Cys peroxiredoxin (2-Cys Prx), which thus plays a key role in maintaining the reducing capacity of chloroplast Trxs in response to light intensity. Based on the severe phenotype of mutant lines lacking NTRC, it is clear that this enzyme plays an essential role in chloroplast redox homeostasis. However, whether the function of NTRC depends on its capacity of reduce 2-Cys Prxs or has additional targets remains unknown. Here, we have addressed this issue by a comparative analysis of the triple mutant of Arabidopsis thaliana, ntrc-2cpab, simultaneously lacking 2-Cys Prxs and NTRC, and the double mutant 2cpab lacking 2-Cys Prxs. The phenotype of the ntrc-2cpab mutant is indistinguishable of the 2cpab mutant, as shown by growth rate, photosynthesis performance, light-dependent redox regulation of enzyme activity and comparative transcriptomics based RNA-Seq. Based on these results, we propose that the function of NTRC in chloroplast redox homeostasis is exerted by the regulation of the redox balance of 2-Cys Prxs rather than by the direct reduction of additional targets.
Project description:Expression analysis of gene expression changes in Homo sapiens SGC-7901 cells after knock down of MTA2 (Metastasis-associated protein) or overexpression SNHG5 (snoRNA host gene 5) Investigation of whole genome gene expression level changes in a Homo sapiens gastric carcinoma cells SGC-7901 after knock down MTA2 expression and upregulation of SNHG5
Project description:EGFR expression in primary mammary tumors predicts for decrease patient surival, but targeted EGFR inhibitors have failed to improve patient outcomes. To identifiy mediators of resistance to EGFR inhibition in breast cancer, normal murine mammary gland (NMuMG) cells specifcally transformed by overexpression of EGFR (NME) were cultured under 3D conditions in the presence of the EGFR inhibitor AG1478. Using this approach we were able to isolate a cell conlony that demonstrated sponaneous resistance to inhibition of EGFR. These AG1478 resistant (AGR) cells were expanded in vitro and analysed by microarray.