ABSTRACT: Whole genome expression profiling of rice leaves treated with Xanthomonas oryzae pv. oryzae secreted cell wall degrading lipase/esteraseA (LipA) at early time points, 30 min. and 120 min.
Project description:Treatment of rice tissues with purified preparations of a Xanthomonas oryzae pv. oryzae (Xoo) secreted plant cell wall degrading enzyme, Lipase/Esterase (LipA), elicits cell wall damage induced innate immune responses. LipA activity is required for induction of defense responses. In order to characterize the early events during elaboration of cell wall degrading enzyme, Lipase/Esterase (LipA) induced innate immune response in rice, we have performed global gene expression profiling of rice leaves treated with purified LipA at early time points, 30 minutes and 120min, after treatment. Whole genome transcriptional profiling was performed using Affymetrix Rice GeneChips Leaves of two weeks old green house grown susceptible rice cultivar (Taichung Native-1; TN-1) were infiltrated with either 30-40μl of purified Xoo Lipase/Esterase (LipA)(500μg/ml) or with buffer (10mM potassium phosphate buffer pH 6.0) alone (as described in Jha et al. 2007; MPMI vol 20, pp 31-40). The plants were shifted to a growth chamber (28oC; 80% relative humidity; 12/12h light/dark cycle) 48 hours before the treatment. 20-30 leaf pieces covering the infiltrated zone from each of the treatments were harvested 30 min. and 120 min. after infiltration. Total RNA isolated from the pooled samples was subjected for expression analysis using Affymetrix GeneChip System. The experiment was repeated with three different biological replicates using RNA isolated from three batches of rice leaves treated with the freshly purified Xoo Lipase/Esterase (LipA)and the buffer Gene Expression profiling of rice leaves undergoing an innate immune respone induced by LipA (Lipase/Esterase A) enzyme
Project description:Treatment of rice tissues with purified preparations of a Xanthomonas oryzae pv. oryzae (Xoo) secreted plant cell wall degrading enzyme, Lipase/Esterase (LipA), elicits cell wall damage induced innate immune responses. LipA activity is required for induction of defense responses. In order to characterize the early events during elaboration of cell wall degrading enzyme, Lipase/Esterase (LipA) induced innate immune response in rice, we have performed global gene expression profiling of rice leaves treated with purified LipA at early time points, 30 minutes and 120min, after treatment. Whole genome transcriptional profiling was performed using Affymetrix Rice GeneChips
Project description:The plant cell wall degrading enzyme LipA (Lipase/Esterase A) is a Type II secretion system secreted protein of Xanthomonas oryzae pv. oryzae (Xoo; the casual of bacterial leaf blight of rice). LipA is an Xoo virulence factor. However, LipA is a double edged sword for Xoo as it induces rice defense responses such as programmed cell death/hypersensitive response like reaction (HR) and callose deposition. Prior treatment with LipA enhances resistance against subseqent Xoo infection. In order to understand the molecular events associated with Esterase (LipA) induced innate immune responsein rice , whole genome transcriptional profiling was performed using Affymetrix Rice GeneChips
Project description:Genes identified in ETEC E24377A after interaction with host cells (Caco-2) after time points of 30 min, 60 min and 120 min. Both the adherent and non-adherent cells were profiled as well as E24377a grown only in tissue culture media.
Project description:Triple-negative breast cancer (TNBC) has a poor clinical outcome, due to a lack of actionable therapeutic targets. Herein we define lysosomal acid lipase A (LIPA) as a viable molecular target in TNBC and identify a stereospecific small molecule (ERX-41) that binds LIPA. ERX-41 induces endoplasmic reticulum (ER) stress resulting in cell death, and this effect is on target as evidenced by specific LIPA mutations providing resistance. Importantly, we demonstrate that ERX-41 activity is independent of LIPA lipase function but dependent on its ER localization. Mechanistically, ERX-41 binding of LIPA decreases expression of multiple ER-resident proteins involved in protein folding. This targeted vulnerability has a large therapeutic window, with no adverse effects either on normal mammary epithelial cells or in mice. Our study implicates a targeted strategy for solid tumors, including breast, brain, pancreatic and ovarian, whereby small, orally bioavailable molecules targeting LIPA block protein folding, induce ER stress and result in tumor cell death.
Project description:We examined the mating response of W303 bar1delta a-type cells to six alpha-factor concentrations (0.06, 0.2, 0.6, 6, 60 and 600 nM). In each alpha factor concentration between five to seven time points were collected. The time points in all experiments (except for concentration 600 nM were time point 15 min was omitted) were 5, 15, 30, 60, and 90 min. In some of the concentrations also time points 120 and 150 min were considered. The expression of cells before and after addition of alpha-factor were compared using S. cerevisiae cDNA microarrays, in all experiments the same sample before adding alpha factor was used as a control. Keywords: yeast mating response, comparison among alpha factor concentrations
Project description:In this project, we follow the temporal proteome dynamics during heat stress response in a model organism Saccharomyces cerevisiae. Using label-free quantification we determine changes in the yeast proteome at different times after mild temperature shift, from 30˚C to 37˚C. The time points measured were 0 min (30˚C, no stress) and then at 10 min, 30 min, 60 min, 120 min and 240 min after cells were transferred to 37 °C. For each time point, four biological replicates were collected for the haploid wild type (BY4742, Mat ALPHA, his3Δ1; leu2Δ0; lys2Δ0; ura3Δ0; YJL088w::kanMX4) strain and a haploid strain with a chaperone Ssb1p deletion (ssb1Δ).
Project description:Previous studies have demonstrated the essential role of morphogenetic regulation in Fusarium oxysporum pathogenesis, including processes such as cell-wall biogenesis, cell division and differentiation of infection-like structures. We identified three F. oxysporum genes encoding predicted transcription factors showing significant identities to Magnaporthe oryzae Con7p, Con7-1, plus two identical copies of Con7-2. Targeted deletion of con7-1 produced non-pathogenic mutants with altered morphogenesis, including defects in cell wall structure, polar growth, hyphal branching and conidiation. By contrast, simultaneous inactivation of both con7-2 copies caused no detectable defects in the resulting mutants. Comparative microarray-based gene expression analysis indicated that Con7-1 modulates the expression of a large number of genes involved in different biological functions, including host-pathogen interactions, morphogenesis and development, signal perception and transduction, transcriptional regulation and primary and secondary metabolism. Taken together, our results points to Con7-1 as general regulator of morphogenesis and virulence in F. oxysporum. Con7-1 dependent gene expression in Fusarium oxysporum wt and M-NM-^Tcon7-1 mutant was measured after shifting the mycelia to PDB pH5.0 for 30 min and the transferred to PDB pH7.0 for 30 min. Three independent experiments were performed.
Project description:We report gene expression in Aspergillus fumigatus mycelium in exposure to either 0.005% ethanol (control) or 5 µg/mL 5,8-dihydroxyoctadecadienoic acid (5,8-diHODE) for 30 min and 120 min
