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Transcriptome analysis of zones of colonies of the ΔflbA strain of Aspergillus niger

ABSTRACT: Gene expression was studied at the periphery, an intermediate zone, and the centre of wild-type and ∆flbA colonies using Affymetrix A. niger whole genome microarrays. We used Affymetrix GeneChip A. niger Geome Arrays and identifed up- and down-regulated genes that may account for the differences between wild-type and ΔflbA colonies.

ORGANISM(S): Aspergillus niger Aspergillus niger CBS 513.88

PROVIDER: GSE44391 | GEO | 2014/02/13

SECONDARY ACCESSION(S): PRJNA189740

REPOSITORIES: GEO

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Transcriptome analysis of zones of colonies of the M-NM-^TflbA strain of Aspergillus niger

Project description:Gene expression was studied at the periphery, an intermediate zone, and the centre of wild-type and M-bM-^HM-^FflbA colonies using Affymetrix A. niger whole genome microarrays. We used Affymetrix GeneChip A. niger Geome Arrays and identifed up- and down-regulated genes that may account for the differences between wild-type and M-NM-^TflbA colonies. RNA was isolated from a biological duplicate of concentric zones of 7-day old wildtype and mutant colonies. Zone 1 represented the most central zone, zone 3 an intermediate zone, and zone 5 the peripheral zone. Mycelium of the distinct zones was harvested from three colonies.

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Project description:Colonies of Aspergillus niger secrete proteins throughout the colony except for the sporulating zone. Strains in which the sporulation gene flbA is deleted do not reproduce asexually and secrete proteins throughout the mycelium. Moreover, ΔflbA hyphae lyse and have thinner cell walls. This pleiotropic phenotype is associated with differential expression of 36 transcription factor genes, of which msnB was inactivated in this study. Whole genome expression analysis of wild-type and ΔmsnB colonies showed differential expression of genes encoding secreted proteins, genes involved in the oxidoreductive balance, and in (secondary) metabolism. Biomass formation, sporulation, and the secretome in the medium were not affected in strain ΔmsnB. In contrast, ΔmsnB secreted more protein when compared to wild type. Taken together, msnB affects protein secretion and is a potential lead for improving A. niger as a cell factory.

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Spatial differentiation of gene expression in Aspergillus niger colonies grown for sugar beet pulp utilization

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The FlbA-regulated predicted transcription factor rpnR of Aspergillus niger is involved in protein secretion and stress resistance

Project description:Proteins are secreted throughout the mycelium of Aspergillus niger except for the sporulating zone. Inactivation of flbA results in colonies that do not reproduce asexually and secrete proteins throughout the colony. Moreover, ΔflbA hyphae lyse and have thinner cell walls. This pleiotropic phenotype is associated with differential expression of 38 transcription factor genes, of which rpnR was inactivated in this study. Biomass formation and sporulation was not affected in strain ΔrpnR but protein secretion was severely reduced. Moreover, ΔrpnR showed decreased resistance to H2O2 and the unfolded protein response inducing agent dithiothreitol. The secretomes of ΔrpnR and wild type consisted of 342 and 367 proteins, respectively. Secretion of several CAZymes and proteins involved in cell wall remodeling was more strongly reduced when compared to other proteins within the secretome of ΔrpnR. Whole genome expression analysis of xylose-grown colonies showed that RpnR activates expression of ribosomal subunits as well as small secreted proteins. On the other hand, it inhibited expression of the genes encoding the arabinofuranosidases AbfA, AbfB and AbfE, as well as genes related to amino acid metabolism, xenobiotics biodegradation and metabolism, lipid metabolism, and metabolism of cofactors and vitamins. Taken together, RpnR is involved in protein synthesis and may also be involved in cell wall architecture. Both functions could explain the reduced protein secretion and proteotoxic stress resistance in the deletion strain.

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colonies

Project description:colonies Metagenome

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Project description:Expression data from batch cultivations of Aspergillus niger wild type strain ATCC 1015 and adrA, facB and creA deletion mutants constructed on ATCC 1015 background strain with glucose or glycerol as carbon sources. Genome-wide transcriptome analysis was used to identify genes either affected directly or indirectly by each transcription factor investigated during growth on a repressing or a derepressing carbon source. For this purpose, batch cultivations under well-controlled conditions were performed with Aspergillus niger wild type strain ATCC 1015 and the three deletion mutants of the corresponding transcription factors AdrA, FacB and CreA. Samples for RNA extraction were collected and further processed for hybridization in custom-designed Affymetrix microarrays containing probes for three Aspergillus species, including A. niger.

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