Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

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

ABSTRACT: 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.

ORGANISM(S): Ustilago maydis

SUBMITTER: Jörg Kämper

PROVIDER: E-GEOD-18750 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

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

Heimel Kai K Scherer Mario M Vranes Miroslav M Wahl Ramon R Pothiratana Chetsada C Schuler David D Vincon Volker V Finkernagel Florian F Flor-Parra Ignacio I Kämper Jörg J

PLoS pathogens 20100805 8

In the phytopathogenic basidiomycete Ustilago maydis, sexual and pathogenic development are tightly connected and controlled by the heterodimeric bE/bW transcription factor complex encoded by the b-mating type locus. The formation of the active bE/bW heterodimer leads to the formation of filaments, induces a G2 cell cycle arrest, and triggers pathogenicity. Here, we identify a set of 345 bE/bW responsive genes which show altered expression during these developmental changes; several of these gen ...[more]

PMID: 20700446

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Project description:We analyzed the transcriptional response of the actinomycete Rhodococcus aetherivorans I24 to biphenyl and polychlorinated biphenyls (PCBs). This species has not been extensively exposed to PCBs, as it was first isolated from a toluene contaminated aquifer, rather than a site contaminated with polychlorinated hydrocarbons. Using a microarray targeting 3524 genes, we assessed gene expression in minimal medium supplemented with various substrates (e.g. PCBs) and in both PCB-contaminated and non-contaminated sediment slurries. Relative to the reference condition (minimal medium supplemented with glucose), 408 genes were up-regulated in the various treatments. In medium and in sediment, PCBs elicited the up-regulation of a common set of 100 genes, including chaperones (groEL), a superoxide dismutase (sodA), alkyl hydroperoxide reductase protein C (ahpC), and a catalase/peroxidase (katG). Analysis of the R. aetherivorans I24 genome sequence identified orthologs of many of the genes in the canonical biphenyl pathway, but very few of these genes were up-regulated in response to PCBs or biphenyl. This study is one of the first which utilizes microarrays to assess the transcriptional response of a soil bacterium to a pollutant under conditions which more closely resemble the natural environment. Our results indicate that the transcriptional response of R. aetherivorans I24 to PCBs, in both medium and sediment, is primarily directed towards reducing oxidative stress, rather than catabolism. In addition, the identification of numerous genes expressed in contaminated soil specifically may have implications for the development of biosensors. Finally, comparative genomic and transcriptomic analyses suggest that the mere presence of orthologs of the required enzymes may not be sufficient to confer a vigorous biphenyl/PCB metabolism. RNA was isolated from cells incubated in the following: sediment from a PCB-contaminated industrial site, uncontaminated sediment from a comparable site, and defined media supplemented with glucose (3 g/L), glucose and biphenyl (3 g/L, 4.5 Î¼M), or glucose and PCBs (3 g/L, 5 mg/L Aroclor 1254). In all cases, there were 3 biological replicates and 2 technical replicates (repeat hybridizations). A total of 3524 genes are represented on the arrays; of these, 41 and 176 are found on the plasmids pRA2 and pRA3, respectively. On average, there are 3 distinct 24nt probes per gene.

Dataset Information

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

Publications

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

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