Dataset Information

Genome-wide transcriptome and proteome analysis of the global GATA transcription factors AreA and AreB in Fusarium fujikuroi

ABSTRACT: We performed genome-wide transcriptome and proteome analyses of the Fusarium fujikuroi wild type and ΔAREA and ΔAREB mutants under nitrogen limitation and excess. Microarray analysis revealed that each transcription factor regulates a total set of about 4200 genes under nitrogen limiting conditions with an enrichment of genes belonging to functional classes of primary/secondary metabolism, transporters, cell defense/virulence and transcription. Under nitrogen excess, AreA is not active as a transcriptional regulator, while AreB affects expression of about 4400 genes. Both transcription factors regulate a common set of about 2100 genes and most of the time work together either as positive or as negative regulators. Our proteomics analysis identified 446 AreA-regulated and 386 AreB-regulated proteins at nitrogen limitation and 138 AreA-regulated and 447 AreB-regulated proteins at nitrogen excess, respectively. Comparison of the proteome and transcriptome data indicates that many post-transcriptional regulations are facilitated by both GATA-factors, for example on enzymes involved in the TCA and glyoxylate cycle and the main glutamate/glutamine cycling.

ORGANISM(S): Fusarium fujikuroi

PROVIDER: GSE80480 | GEO | 2017/05/10

SECONDARY ACCESSION(S): PRJNA319032

REPOSITORIES: GEO

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Project description:"Background: The ascomycete Fusarium fujikuroi causes “bakanae” disease of rice due to its ability to produce gibberellins (GA), but is also known for producing a broad spectrum of other secondary metabolites, (SM) such as pigments and harmful mycotoxins. Biosynthesis of multiple SM is regulated by nitrogen availability. The global GATA transcription factors (TF) AreA and AreB were shown to play key roles in this nitrogen-dependent regulation. Recently, we have shown that AreA activates the transcription of AREB, but not vice versa. Both regulators interact in the nucleus, indicating a complex regulatory interplay between both regulators. Results: We performed transcriptome and proteome analyses of the wild type and ΔAREA and ΔAREB mutants under nitrogen limiting and sufficient conditions. Microarray analysis revealed that each transcription factor regulates a total set of about 4,200 genes under nitrogen limiting conditions. Gene enrichment analysis revealed that significantly up- and down-regulated genes belong to functional classes of primary/secondary metabolism, transporters, cell defense/virulence and transcription. Under nitrogen excess, AreA is not active as a transcriptional regulator, while AreB affects expression of about 4,400 genes. Both transcription factors regulate a common set of about 2,100 genes, often coordinately either as positive or as negative regulators. Our proteomics analysis identified 446 AreA-regulated and 386 AreB-regulated proteins under nitrogen limitation and 138 AreA-regulated and 447 AreB-regulated proteins under nitrogen sufficiency, respectively. Comparison of the proteome and transcriptome data indicates that both GATA-factors not only regulate expression of target genes but also affect protein abundance by post-transcriptional regulations, for example on enzymes involved in glutamate cycling, as well as histone acetylation at some of the target SM gene clusters. Conclusion: This study provides novel insights into the AreA- and AreB-mediated regulation network on a cellular scale. Microarray analysis clearly demonstrated that both GATA-factors are major regulators of alternate nitrogen assimilatory pathways and secondary metabolism. Moreover, proteome analysis of the AREA and AREB deletion mutants showed that both TF are global players interconnecting carbon and nitrogen regulation. They mainly act as mutualistic, but also antagonistic regulators and operate by transcriptional and post-transcriptional regulations and epigenetic modifications." (Abstract taken from manuscript)

Project description:Nitrogen is one of the major nutrients limiting microbial productivity in the ocean, and as a result marine microorganisms have evolved specialized systems for responding to nitrogen stress. The highly abundant alphaproteobacterium Candidatus Pelagibacter ubique lacks the canonical GlnB, GlnD, and NtrB/NtrC genes for regulating nitrogen assimilation. A survey of 127 Alphaproteobacteria genomes found these genes to be highly represented in free-living and pathogenic organisms with large genomes and only missing in a subset of obligate intracellular organisms and other SAR11 strains. We examined global differences in mRNA and protein expression in Ca. P. ubique strain HTCC1062 during nitrogen-limited and nitrogen-replete stationary phase to understand how this thriving organism responds to nitrogen limitation. Transporters for ammonium (AmtB), taurine (TauA), amino acids (YhdW), and opines (OccT) were all elevated in nitrogen-limited cells, indicating they devote increased resources to the assimilation of nitrogenous compounds. Enzymes for assimilating amine into glutamine (GlnA) and glutamate (AspC, GltBD) were similarly up-regulated. Differential regulation of the transcriptional regulator NtrX in the two-component signaling system NtrY/NtrX was also observed, implicating it in the control of the nitrogen starvation response. Comparisons of the transcriptome and proteome suggest that Amt is post-transcriptionally repressed during nitrogen limitation, supporting previous studies that computationally identified a novel cis-acting riboswitch upstream of this gene. These observations support the conclusion that Ca. P. ubique has an unusually simple regulatory system that enables it to increase its capacity for the uptake of nitrogenous compounds in response to nitrogen limitation. Batch cultures of P. ubique were grown in a defined arificial seawater media. Three cultures were given no nitrogen amendment, and three other cultures received an excess concentration of NH3. Cultures were harvested for microarray analyses during log and stationary phase for the purpose of observing differences in gene expression related to nitrogen limitation. Proteomic analysis was conducted in parallel and is available at http://omics.pnl.gov .

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Genome-wide transcriptome and proteome analysis of the global GATA transcription factors AreA and AreB in Fusarium fujikuroi

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