Analysis of LOV-FPs-mediated stress response in E. coli.
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ABSTRACT:
ORGANISM(S): Gluconobacter oxydans Escherichia coli Corynebacterium glutamicum Escherichia coli str. K-12 substr. MG1655 Bacillus subtilis subsp. subtilis str. 168
PROVIDER: GSE110168 | GEO | 2018/10/09
REPOSITORIES: GEO
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Project description:We identified a small zinc finger protein, MBS, as a new mediator of singlet oxygen responses in Chlamydomonas and Arabidopsis. MBS is required for induction of singlet oxygen-dependent gene expression and, upon oxidative stress, accumulates in distinct granules in the cytosol of Arabidopsis cells. First, we recorded changes in light stress-regulated gene expression profiles after genetically perturbing MBS function by isolating mutants for the two MBS genes (MBS1 and MBS2) and by overexpression of MBS1 in Arabidopsis thaliana. Then, these light stress-related gene expression profiles were analyzed with respect to genes specifically responding to singlet oxygen and hydrogen peroxide/superoxide. The results indicated that MBS inactivation leads to an impaired response to singlet oxygen signaling under light stress.
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Project description:Analysis of LOV-FPs-mediated stress response in E. coli.
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Project description:The facultatively photosynthetic bacterium Rhodobacter sphaeroides harbors an unusual LOV (light, oxygen, voltage) domain protein, RsLOV. While showing a characteristic photocycle, the protein misses a C - terminal output domain, similar to PpSB2 in Pseudomonas putida. Oxygen tension and light quantity are the two main responsible factors controlling the expression of photosynthesis genes in Rhodobacter sphaeroides. Two photoreceptor proteins are known to be involved in this regulation: the intensively studied AppA protein and the more recently identified cryptochrome-like protein CryB. Here we show by transcriptome and physiological studies that RsLOV is also involved in the regulation of photosynthetic gene expression. Our data further hint to a connection between RsLOV and the carbon hydrate metabolism, chemotaxis, as well as to the cellular response to photooxidative stress. RsLOV does not only affect blue light dependent gene expression but also redox-dependent regulation. This SuperSeries is composed of the following subset Series: GSE33194: R. sphaeroides ?lov vs. R. sphaeroides 2.4.1 (microarobic conditions) GSE33259: R. sphaeroides ?lov vs. R. sphaeroides 2.4.1 (blue light, semiaerobic conditions) GSE33260: R. sphaeroides ?lov vs. R. sphaeroides 2.4.1 (singlet oxygen stress, aerobic conditions)
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Project description:The goal of this work was to investigate oxidative stress responses of Arabidopsis to low red to far-red ratios of light as a signal of competition using a biological weedy and an artificial source of far-red light. More specifically, elucidation of the signaling role of singlet oxygen in Arabidopsis under low red-to far-red light environments was the major objective of this work. Oxidative stress responses of Arabidopsis to low red (R) to far-red (FR) signals (R:FR ≈ 0.3), generated by a biological weedy and an artificial source of FR light, were compared with a weed-free control (R:FR ≈1.4). In the low R:FR treatments, induction of the shade avoidance responses coincided with increased singlet oxygen (1O2) production and decreased level of superoxide and superoxide dismutase activity. Although the increase of 1O2 was not due to protochlorophyllide accumulation and did not result in cell death, treatments with the 1O2 generator 5-aminolevulinic acid increased sensitivity to cell death. Transcriptome responses minimally resembled those reported in four Arabidopsis 1O2 generating systems such that only few genes (6 out of 1931) were consistently up-regulated supporting the specificity of 1O2 signaling. Moreover, suppressors of jasmonate accumulation, including the 1O2-responsive amidohydrolase ILL6, the sulfotransferase ST2a, which are involved in prioritization of elongation growth versus defense were consistently up-regulated. Our data support a model in which photoreceptors connect low R:FR light cues to the JA signaling pathway. Repression of bioactive JAs via the amidohydrolase ILL6, and sulfotransferase ST2a may promote the shade avoidance (versus defense) and 1O2 acclimation (versus cell death) responses to competition cues.
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Project description:We used the flu mutant of Arabidopsis to detail gene expression in response to singlet oxygen. The conditional flu mutant of Arabidopsis accumulates excess protochlorophyllide in the dark within chloroplast membranes that upon illumination acts as a photosensitizer and generates singlet oxygen. Immediately after the release of singlet oxygen mature flu plants stop growing, whereas seedlings bleach and die. Within the first 30 min after the release of singlet oxygen rapid changes in nuclear gene expression occur. Distinct sets of genes were activated that were different from those induced by other reactive oxygen species, superoxide or hydrogen peroxide. Experiment Overall Design: Arabidopsis thaliana rosette leaves were harvested after 30 min, 1h, and 2 h of reillumination following a 8h dark period for RNA extraction and hybridization on Affymetrix ATH1 microarrays. Plants were grown on soil for 3 weeks under continuous light at 90 mmol. m-2 . s-1. For each sample, the rosette leaves of five to six 3-week-old plants (before they start bolting) were collected for RNA extraction. Total RNAs from two separate biological experiments were pooled for the preparation of cDNA and the subsequent synthesis of biotin-labeled complementary RNA as recommended by Affymetrix.
2008-04-05 | E-GEOD-10876 | biostudies-arrayexpress
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Project description:We identified a small zinc finger protein, MBS, as a new mediator of singlet oxygen responses in Chlamydomonas and Arabidopsis. MBS is required for induction of singlet oxygen-dependent gene expression and, upon oxidative stress, accumulates in distinct granules in the cytosol of Arabidopsis cells. First, we recorded changes in light stress-regulated gene expression profiles after genetically perturbing MBS function by isolating mutants for the two MBS genes (MBS1 and MBS2) and by overexpression of MBS1 in Arabidopsis thaliana. Then, these light stress-related gene expression profiles were analyzed with respect to genes specifically responding to singlet oxygen and hydrogen peroxide/superoxide. The results indicated that MBS inactivation leads to an impaired response to singlet oxygen signaling under light stress. Knock-out or knock-down of the MBS1 and MBS2 genes and overexpression of MBS1 in Arabidopsis thaliana were compared with the wild type and the flu mutant as controls under light stress. Arabidopsis seedlings of mbs1-1, RNAi-MBS2/mbs1-1, 35S:MBS1, the wild type and the flu mutant were harvested at 0 h and 3 h of light stress (HL, 1000 µE m−2 s−1). Total RNA was extracted and cDNA was hybridized to Affymetrix ATH1 microarrays in three biological replicates.
2014-04-21 | E-GEOD-49596 | biostudies-arrayexpress
Project description:Purpose: Avian photoreceptors are a diverse class of neurons, comprised of four single cones, the two members of the double cone, and rods. Many distinctive features of photoreceptor subtypes, including spectral tuning, oil droplet size and pigmentation, synaptic targets and spatial patterning, have been well characterized, but the molecular mechanisms underlying these attributes have not been explored. Furthermore, the signaling events and transcriptional regulators driving the differentiation of these diverse photoreceptors are currently unknown. Methods: To identify genes specifically expressed in distinct chicken (Gallus gallus) photoreceptor subtypes, we developed fluorescent reporters that label photoreceptor subpopulations, isolated these subpopulations using fluorescence-activated cell sorting, subjected them to next-generation sequencing, and conducted differential expression analysis. Results: We identified hundreds of differentially expressed genes from photoreceptor subpopulations labeled with rhodopsin, red opsin, green opsin, and violet opsin reporters. These genes are involved in a variety of processes, including phototransduction, transcriptional regulation, cell adhesion, maintenance of intra- and extra-cellular structure, and metabolism. Of particular note are a variety of differentially expressed transcription factors, which may drive and maintain photoreceptor diversity, and cell adhesion molecules that may mediate spatial patterning of photoreceptors and act to establish retinal circuitry. Conclusions: These analyses provide a framework for future studies that will dissect the role of these various factors in the differentiation of avian photoreceptor subtypes. mRNA expression profiling of 5 pairs of photoreceptor subtypes isolated from chicken retinal explants, 3 replicates per sample
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