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Comparison of expression profiles for Agrobacterium tumefaciens wild-type strain C58 grown with or without manganese

ABSTRACT: The plant pathogen Agrobacterium tumefaciens attaches to and forms biofilms on both biotic and abiotic surfaces. The transition between free-living, planktonic A. tumefaciens and multicellular biofilms is regulated by several well-defined environmental and nutritional inputs, including pH, oxygen tension, and phosphate concentration. In many bacterial species limiting iron levels inhibit attachment and biofilm formation. In several systems intracellular levels of the redox-active manganous (Mn2+) and ferrous (Fe2+) ions are interrelated and have tight corresponding regulation with respect to one another. We show that limiting manganese concentrations elicit similar growth and biofilm phenotypes to those seen under iron-limiting conditions. Microarray analysis comparing gene expression in manganese-replete versus manganese-limiting conditions identified a small number of differentially regulated transcripts. These results indicate that the redox-active manganous and ferrous ions are required for wild-type levels of growth and biofilm formation, and that the manganese-dependent response is primarily post-transcriptional and complementary to, but not redundant with, the iron-dependent response. Four biological replicates, independent RNA preparations, two dye swaps.

ORGANISM(S): Agrobacterium tumefaciens

SUBMITTER: Jason Heindl

PROVIDER: E-GEOD-71266 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Comparison of expression profiles for Agrobacterium tumefaciens wild-type strain C58 grown with or without iron.

Project description:The plant pathogen Agrobacterium tumefaciens attaches to and forms biofilms on both biotic and abiotic surfaces. The transition between free-living, planktonic A. tumefaciens and multicellular biofilms is regulated by several well-defined environmental and nutritional inputs, including pH, oxygen tension, and phosphate concentration. In many bacterial species limiting iron levels inhibit attachment and biofilm formation. We demonstrate that A. tumefaciens biofilm formation is reduced under limiting iron conditions. Treatment of A. tumefaciens cultures with EDDHA, an iron-specific extracellular chelator, inhibited both planktonic growth rate and adherent biomass. These effects were reversed upon addition of exogenous ferrous iron. This reduced biofilm formation effect is independent of the known iron-responsive regulators Irr and RirA. Transcriptome analysis comparing gene expression under iron-replete versus iron-deficient conditions identified hundreds of genes that are differentially regulated. Downregulated genes suggest an iron sparing response. Four biological replicates, independent RNA preparations, one dye swap.

2015-07-22 | E-GEOD-71138 | biostudies-arrayexpress

Comparison of expression profiles for Agrobacterium tumefaciens wild-type strain C58 grown with or without manganese

Project description:The plant pathogen Agrobacterium tumefaciens attaches to and forms biofilms on both biotic and abiotic surfaces. The transition between free-living, planktonic A. tumefaciens and multicellular biofilms is regulated by several well-defined environmental and nutritional inputs, including pH, oxygen tension, and phosphate concentration. In many bacterial species limiting iron levels inhibit attachment and biofilm formation. In several systems intracellular levels of the redox-active manganous (Mn2+) and ferrous (Fe2+) ions are interrelated and have tight corresponding regulation with respect to one another. We show that limiting manganese concentrations elicit similar growth and biofilm phenotypes to those seen under iron-limiting conditions. Microarray analysis comparing gene expression in manganese-replete versus manganese-limiting conditions identified a small number of differentially regulated transcripts. These results indicate that the redox-active manganous and ferrous ions are required for wild-type levels of growth and biofilm formation, and that the manganese-dependent response is primarily post-transcriptional and complementary to, but not redundant with, the iron-dependent response.

2015-07-24 | GSE71266 | GEO

Comparison of expression profiles for Agrobacterium tumefaciens wild-type strain C58 grown with or without iron.

2015-07-22 | GSE71138 | GEO

Comparison of expression profiles for Agrobacterium tumefaciens wild-type strain C58 and isogenic pdhS2 (Atu1888) deletion strain

Project description:A conserved regulatory pathway directs developmental transitions and asymmetries in Agrobacterium tumefaciens. Core components of this coordination of division and development (CDD) pathway include two integrated phosphorelays. One of these phosphorelays includes at least four histidine sensor kinase homologues, DivJ, PleC, PdhS1, and PdhS2, and at least two response regulators, DivK and PleD. Previously we demonstrated that PdhS2 reciprocally regulates biofilm formation and swimming motility. In the current study we further dissect the architecture of the CDD pathway in A. tumefaciens with respect to PdhS2. We show that PdhS2-dependent effects on attachment and motility require the response regulator, DivK, but do not require PdhS2 autokinase or phosphotransfer activities. We also demonstrate that PdhS2 regulation of biofilm formation is dependent upon multiple diguanylate cyclases, including PleD, DgcA, and DgcB, implying that PdhS2 regulation of this process intersects with pathways regulating levels of the second messenger cyclic-di-GMP. Finally, we show that upon cell division a GFP fusion to PdhS2 dynamically localizes to the new pole of the bacterium suggesting that PdhS2 controls processes in the daughter cell compartment of predivisional cells. These observations suggest that PdhS2 negatively regulates DivK, and possibly PleD, activity to control developmental processes in the daughter cell compartment of predivisional cells, as well as in newly released motile daughter cells. Three biological replicates, independent RNA preparations, one dye swap.

2015-07-24 | E-GEOD-71267 | biostudies-arrayexpress

Comparison of Expression Profiles for Agrobacterium tumefaciens visR mutant to wild-type strain.

Project description:Many bacteria colonize surfaces and transition to a sessile mode of growth. The plant pathogen Agrobacterium tumefaciens produces a unipolar polysaccharide (UPP) adhesin at single cell poles that contact surfaces. Here we report that elevated levels of the intracellular signal cyclic diguanosine monophosphate (c-di-GMP) lead to surface-contact-independent UPP production and a red colony phenotype due to production of UPP and the exopolysaccharide cellulose, when A. tumefaciens is incubated with the polysaccharide stain Congo Red. Transposon mutations with elevated Congo Red staining identified presumptive UPP-negative regulators, mutants for which were hyperadherent, producing UPP irrespective of surface contact. Multiple independent mutations were obtained in visN and visR, activators of flagellar motility in A. tumefaciens, now found to inhibit UPP and cellulose production. Expression analysis in a visR mutant and isolation of suppressor mutations, identified three diguanylate cyclases inhibited by VisR. Null mutations for two of these genes decrease attachment and UPP production, but do not alter cellular c-di-GMP levels. However, analysis of catalytic site mutants revealed their GGDEF motifs are required to increase UPP production and surface attachment. Mutations in a specific presumptive c-di-GMP phosphodiesterase also elevate UPP production and attachment, consistent with c-di-GMP activation of surface-dependent adhesin deployment. Three biological replicates, independent RNA preparations, one dye swap.

2014-08-21 | E-GEOD-60565 | biostudies-arrayexpress

Comparison of Expression Profiles for Agrobacterium tumefaciens exoRexoA double mutant to exoA mutant

Project description:Agrobacterium tumefaciens is a facultative plant pathogen and the causative agent of crown gall disease. The initial stage of infection involves attachment to plant tissues and, subsequently, biofilms may form at these sites. This study focuses on the periplasmic ExoR regulator, which was identified based on the severe biofilm deficiency of A. tumefaciens exoR mutants. Genome-wide expression analysis was performed to elucidate the complete ExoR regulon. Overproduction of the exopolysaccharide succinoglycan is a dramatic phenotype of exoR mutants. Comparative expression analyses revealed that the core ExoR regulon is unaffected by succinoglycan synthesis. Several findings are consistent with previous observations: genes involved in succinoglycan biosynthesis, motility, and type VI secretion are differentially expressed in the ?exoR mutant. In addition, these studies revealed new functional categories regulated by ExoR including genes related to virulence, conjugation of the pAtC58 megaplasmid, ABC transporters, and cell envelope architecture. To address how ExoR exerts a broad impact on gene expression from its periplasmic location, a genetic screen was performed to isolate suppressor mutants that mitigate the exoR motility phenotype and identify downstream components of the ExoR regulatory pathway. This suppression analysis identified the acid-sensing two-component system ChvG-ChvI, and the suppressor mutant phenotypes suggest that all or most of the characteristic exoR properties are mediated through ChvG-ChvI. Subsequent analysis indicates that exoR mutants are simulating a response to acidic conditions, even in neutral media. This work expands the model for ExoR regulation in A. tumefaciens and underscores the global role this regulator plays on gene expression. Three biological replicates, Independent RNA preparations, One dye-swap

2014-07-26 | E-GEOD-59789 | biostudies-arrayexpress

Comparison of Expression Profiles for Agrobacterium tumefaciens ExoR mutant to Wild Type

Project description:Agrobacterium tumefaciens is a facultative plant pathogen and the causative agent of crown gall disease. The initial stage of infection involves attachment to plant tissues and, subsequently, biofilms may form at these sites. This study focuses on the periplasmic ExoR regulator, which was identified based on the severe biofilm deficiency of A. tumefaciens exoR mutants. Genome-wide expression analysis was performed to elucidate the complete ExoR regulon. Overproduction of the exopolysaccharide succinoglycan is a dramatic phenotype of exoR mutants. Comparative expression analyses revealed that the core ExoR regulon is unaffected by succinoglycan synthesis. Several findings are consistent with previous observations: genes involved in succinoglycan biosynthesis, motility, and type VI secretion are differentially expressed in the ΔexoR mutant. In addition, these studies revealed new functional categories regulated by ExoR including genes related to virulence, conjugation of the pAtC58 megaplasmid, ABC transporters, and cell envelope architecture. To address how ExoR exerts a broad impact on gene expression from its periplasmic location, a genetic screen was performed to isolate suppressor mutants that mitigate the exoR motility phenotype and identify downstream components of the ExoR regulatory pathway. This suppression analysis identified the acid-sensing two-component system ChvG-ChvI, and the suppressor mutant phenotypes suggest that all or most of the characteristic exoR properties are mediated through ChvG-ChvI. Subsequent analysis indicates that exoR mutants are simulating a response to acidic conditions, even in neutral media. This work expands the model for ExoR regulation in A. tumefaciens and underscores the global role this regulator plays on gene expression. Four biological replicates, Independent RNA preparations, Two dye-swaps

2014-07-26 | E-GEOD-59788 | biostudies-arrayexpress

Study physiological and biochemical response of Phaeocystis antarctica to different irradiance and manganese/ iron deficiency

Project description:Iron and light are typically recognized as major limiting factors controlling phytoplankton growth in the Southern Ocean. Recent field-based evidence suggests, however, that manganese concentrations in this region can be low enough to impact phytoplankton physiology and primary productivity. Our study examined the interactive influence of combined iron and manganese deprivation on protein expression and photophysiology in Phaeocystis antarctica, a key Antarctic phytoplankter, and provide taxon-specific proteomic evidence that natural Southern Ocean Phaeocystis populations regularly experience stress due to combined low manganese and iron availability. In culture, combined low iron and manganese induced large scale changes in the Phaeocystis proteome and resulted in reorganization of key components of the photosynthetic apparatus; these differences were largely distinct from those arising from changes in irradiance. These results implicate manganese availability as an important driver of Southern Ocean productivity and demonstrate the utility of peptide mass spectrometry as a tool for mapping of manganese contributions to HNLC conditions in this region.

2019-06-25 | PXD010974 | Pride

Gene expression profiling of 3T3 fibroblast cells cultured with degradable metallic materials

Project description:Metals have been used for coronary artery stent materials in order to prevent stenosis caused by atherosclerosis. Degradable metallic materials (DMMs) are considered to be useful to avoid in-stent restenosis and late thrombosis. A new DMM, Fe-35Mn alloy, was fabricated through powder metallurgy in order to satisfy the ideal criteria of cardiovascular stent made of DMM. Since in-stent restenosis is mediated by the extracellular matrix production, which is mostly regulated by fibroblasts, the gene expression profile of 3T3 fibroblasts in the presence of Fe-35Mn alloy was then investigated. The mechanism of cellular responses in the presence of DMM is then expected to be clearly described through the gene profiling experiment. 3T3 fibroblast cells derived from mouse (Mus musculus) embryo (BALB/3T3 clone A31, ATTC) were put in the culture and treated with iron, manganese, and Fe-35Mn alloy for 24 hour in parallel and followed by the RNA extraction. Six technical replicates were included for treated cells as well as control cells.

2010-07-24 | E-GEOD-22794 | biostudies-arrayexpress

Gene expression profiling of SHSY5Y cells exposed to manganese

Project description:The trace element manganese is essential for normal development for all the organisms. Overexposure of manganese may leads to multiple neuronal disorders such as Parkinson, manganism. To explore the molecular mechanism of manganese induced neurotoxicity, gene expression profiling was performed on human neuroblastoma SHSY-5Y cells. Cells were exposed to sub-lethal concentration of manganese (100 Î¼M) for 24 hrs. Our result demonstrates that manganese alter multiple biological pathways including chromatin assembly, neurogenesis and apoptotic pathways.Cells grown in 75mm2 flask. three replicates for each sample SHSY5Y cells grown in MEM:F12 media (1:1) were treated with manganese and total RNA was isolated from cells after 24 hour exposure Three replicate were used for the experiment

2016-07-01 | E-GEOD-70845 | biostudies-arrayexpress

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