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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Effect of clp1 induction on b-dependent gene regulation

ABSTRACT: The clp1 gene is a b-regulated gene that is required for clamp formation during the biotrophic growth phase of U. maydis. Clp1 expression blocks b-dependent filament formation and cell cycle arrest if co-induced with the b-heterodimer. Strains AB33 (control) and UMS84 were grown for the times indicated in array medium containing 3 g/l nitrate and 1% arabinose for the induction of nar1P- (bE1/bW2) and crg1P- (clp1) controlled gene expression. Samples were taken from two biological replicates each.

ORGANISM(S): Ustilago maydis

SUBMITTER: Jörg Kämper

PROVIDER: E-GEOD-21119 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Transcription factor Rbf1 is the master regulator for b-mating type controlled pathogenic development in Ustilago maydis

Project description:This SuperSeries is composed of the following subset Series: GSE18750: Controlled expression of compatible and incompatible combinations of Ustilago maydis b-mating type locus genes bE and bW GSE18754: Effect of rbf1 deletion during controlled expression of of Ustilago maydis b-mating type locus genes bE1 and bW2 GSE18756: Rbf1 induced gene expression in Ustilago maydis Refer to individual Series

2010-06-01 | E-GEOD-18881 | biostudies-arrayexpress

Controlled expression of compatible and incompatible combinations of Ustilago maydis b-mating type locus genes bE and bW

Project description:In the phytopathogenic Basidiomcete Ustilago maydis, sexual and pathogenic development are tightly connected and controlled via the b-mating type locus. The b-mating type locus encodes two homeodomain transcription factors, bE and bW, which form an active heterodimeric complex when they are derived from different b-alleles. To identify genes regulated by the bE/bW transcription factor, changes in gene expression were monitored during a 12-h time course (0h, 1h, 2h, 3h, 5h, 12h) in strains AB31 and AB33 that harbor the active combination bE1 and bW2 under the control of the arabinose-inducible crg1 promoter and the nitrate-inducible nar1 promoter, respectively (Brachmann et al., 2001, Mol Microbiol 42, 1047-1063). Induction of bE1/bW2 in these strains results in a filament that resembles the infectious dikaryotic hypha formed after fusion of compatible sporidia. Strains AB32 and AB34, which harbor the incompatible bE2 and bW2 combination, were used as controls. Strains were grown to an OD600 of 0.4-0.6 at 28°C in liquid array medium: 6.25% (w/v) salt solution, 30 mM L-glutamine, 1% (w/v) glucose, pH 7.0 (filter-sterilized). For induction of genes under control of the crg1 promoter (strains AB31 and AB32) cells were inoculated in liquid array medium containing 1% (w/v) arabinose instead of glucose as a carbon source. For induction of the nar1-promoter glutamine was exchanged by nitrate (3g KNO3/l) as the sole nitrogen source. For induction, cells were washed once with inducing medium; time point 0 h of the timecourse experiments corresponds to the first contact with inducing medium. Experiments were performed in two biological replicates for each time point.

2010-06-01 | E-GEOD-18750 | biostudies-arrayexpress

Effect of clp1 induction on rbf1-dependent gene regulation

Project description:The clp1 gene is a b-regulated gene that is required for clamp formation during the biotrophic growth phase of U. maydis. Interaction studies demonstrated that Clp1 imnteracts with Rbf1, the central regulator of pathogenciity. To exclude b-dependent effects the experiments were performed in a b-null background to analyze clp1-depndent effects on rbf controlled gene regulation Strains UKH156 (nitrate inducible Rbf1 expression) and UKH164 (arabinose-inducible Clp1 and and nitrate-inducible Rbf1 expression) were grown for 12h in liquid array medium/nitrate/arabinose.

2010-09-17 | E-GEOD-21121 | biostudies-arrayexpress

Effect of rbf1 deletion during controlled expression of Ustilago maydis b-mating type locus genes bE1 and bW2

Project description:In the phytopathogenic Basidiomcete Ustilago maydis, sexual and pathogenic development are tightly connected and controlled via the b-mating type locus. The b-mating type locus encodes two homeodomain transcription factors, bE and bW, which form an active heterodimeric complex when they are derived from different b-alleles. Rbf1 encodes a zinc-finger transcription factor that is expressed upon formation of an active bE/bW heterodimer. To analyse the dependency of b and rbf1, changes in gene expression were monitored in strain AB31 (Brachmann et al., 2001, Mol Microbiol 42, 1047-1063) and in the derivative AB31Δrbf1 in which rbf1 was deleted. AB31 harbours the active combination bE1 and bW2 under the control of the arabinose-inducible crg1 promoter. Samples were taken 3h, 5h and 12h after induction of bE/bW gene expression. Strain AB32, which harbors the incompatible bE2 and bW2 combination, was used as control. Strains were grown to an OD600 of 0.4-0.6 at 28°C in liquid array medium: 6.25% (w/v) salt solution, 30 mM L-glutamine, 1% (w/v) glucose, pH 7.0 (filter-sterilized). For induction of the bE and bW genes, cells were inoculated in liquid array medium containing 1% (w/v) arabinose instead of glucose as a carbon source. For induction, cells were washed once with inducing medium; time point 0 h of the time course experiments corresponds to the first contact with inducing medium. Experiments were performed in two biological replicates for each time point.

2010-06-01 | E-GEOD-18754 | biostudies-arrayexpress

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Project description:Adr1 of Ustilago maydis is a protein kinase that is activated after separation from the regulatory subunit mediated by high cAMP levels. A copy of the gene under the control of the arabinose-inducible crg1 promotor was introduced into the Cbx locus of the wild type strain FB1 creating strain HE140. To monitor the transcriptional changes upon adr1 induction by arabinose, we conducted a time course experiment comparing the wild type transcriptional response before and 75 or 180 min after medium shift to complete medium containing arabinose, as well as the response of the mutant HE140 before and 75 or 180 min after induction of the additional copy of adr1 by arabinose. Keywords: gene induction Strains FB1 (control) and HE140 (a1b1 ipr[pcrg1:adr1]ips) were grown in CM medium containing glucose and were shifted to CM medium containing arabinose and grown for 75 or 180 min. Samples were taken before and after arabinose induction from two (glucose) or three (arabinose) biological replicates each.

2010-06-12 | E-GEOD-6039 | biostudies-arrayexpress

Gene expression on different carbon sources in response to hxt1 deletion in Ustilago maydis

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2009-09-29 | E-GEOD-16634 | biostudies-arrayexpress

ChIP-seq analysis of the Cib1 transcription factor with and without clp1 overexpression.

Project description:For ChIPseq analyses JB1 and UVO151 (JB1 Pcrg:clp1) were transformed with a Cib1-3xHA fusion construct was expressed under control of the endogenous promoter. The fusion construct was integrated at the endogenous locus replacing the native Cib1 gene.

2019-04-04 | E-MTAB-7460 | biostudies-arrayexpress

RNAseq of Ustilago maydis with inducible clp1 and Δcib1

Project description:Ustilago maydis strain JB1 was modified to include an arabinose inducible clp1 in the ip-locus and clb1 was deleted.

2019-04-04 | E-MTAB-7463 | biostudies-arrayexpress

Utilisation of naphthalene by Pseudomonas putida CSV86 under nitrogen varying conditions

Project description:The response of P. putida CSV86 in response to preferential carbon source and varying nitrogen levels was studied in this experiment. The culture was grown in minimal medium with either naphthalene (0.1%) alone or along with glucose (0.25%) as carbon source and different concentration of ammonium nitrate. For nitrogen limiting 0.1% NH4N03 was added in the medium and for nitrogen rich 1% NH4N03 was used. As control CSV86 was also grown in 1X Luria Broth with glucose as additional carbon source. Overall it was seen that the strain could grow efficiently in varying nitrogen conditions by adapting or regulating their metabolism without compromising on the growth rate. To ascertain how the cells were coping with the nitrogen stress, gene expression by microarray was performed. RNA extraction was done using Qiagen RNeasy minikit (Germany). Standard Affymetrix protocol was followed for hybridization with GeneChip P. aeruginosa Genome Array supplied by Affymetrix as GeneChip for P. putida were not supplied by any company.

2014-04-24 | E-MTAB-2528 | biostudies-arrayexpress

Transcriptional comparison of the filamentous fungus Neurospora crassa growing on three major monosaccharides D-glucose, D-xylose and L-arabinose

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