Deletion of the FlbA-regulated transcription factor msnB of Aspergillus niger affects protein secretion
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PROVIDER: PRJNA413426 | ENA |
REPOSITORIES: ENA
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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.
2018-10-01 | GSE104669 | GEO
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.
2018-06-01 | GSE102899 | GEO
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. Triplicate batch fermentations of each of the four Aspergillus niger strains used, the wild type A. niger strain ATCC 1015 and three gene deletion mutants, were carried out using glucose or glycerol as carbon source, and transcriptome analysis was performed. Biomass from each batch cultivation was harvested in the exponential phase of growth and further processed for RNA extraction and hybridization on Affymetrix microarrays.
2011-01-31 | E-GEOD-19952 | biostudies-arrayexpress
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.
2011-01-31 | GSE19952 | GEO
Project description:Aspergillus niger fulfills an important role in nature in nutrient cycling and is a main cell factory for the industrial production of metabolites and enzymes. Inactivation of flbA in A. niger results in thinner cell walls, increased cell lysis, abolished sporulation, and an increased secretome complexity. A total of 36 transcription factor genes are differentially expressed in ∆flbA. Here, seven of these genes (abaA, aslA, aslB, azf1, htfA, nosA, and srbA) were inactivated. Inactivation of each of the genes affected sporulation. Moreover, inactivation of each of the genes impacted protein secretion with the exception of abaA. The impact on secretion was strongest in the case of ΔaslA and ΔaslB that showed increased pepsin, cellulase, and amylase activity. Biomass formed on agar plates was reduced when either abaA, aslA, nosA, or srbA were inactivated. In contrast, biomass of ∆aslA and ∆aslB was higher in liquid shaken cultures. The ∆aslA and ∆htfA strains showed reduced resistance to H2O2, while ∆abaA showed increased resistance to cell wall perturbing agents. By contrast, AslA, AslB, HtfA, Azf1, NosA, and SrbA were shown to be important for cell wall integrity. Together, the seven genes downstream of FlbA impact biomass formation, sporulation, protein secretion, and stress resistance in A. niger, and thereby explain part of the pleiotropic phenotype of the ∆flbA strain.
2024-05-15 | GSE254305 | GEO
Project description:This approach aims at searching unidentified regulatory roles of the AreB transcription factor in the overall carbon metabolism of A. niger. A full areB gene deletion mutant was constructed and characterized in A. niger ATCC 1015. Both strains were grown on glucose or glycerol using ammonia as nitrogen source in batch cultivations and the transcriptome was analyzed using three biological replicated transcriptome experiments. Two areB gene deletion replicates, one on glucose and one on glycerol were discarded due to bad quality and therefore not included in the analysis. 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. Triplicate batch fermentations with the two Aspergillus niger strains used, the wild type A. niger strain ATCC 1015 and the areB complete gene deletion strain were carried out and transcriptome analysis was performed. Biomass from each batch cultivation was harvested in the exponential phase of growth and further processed for RNA extraction and hybridization on Affymetrix microarrays.
2011-01-31 | E-GEOD-21015 | biostudies-arrayexpress
Project description:This approach aims at searching unidentified regulatory roles of the AreB transcription factor in the overall carbon metabolism of A. niger. A full areB gene deletion mutant was constructed and characterized in A. niger ATCC 1015. Both strains were grown on glucose or glycerol using ammonia as nitrogen source in batch cultivations and the transcriptome was analyzed using three biological replicated transcriptome experiments. Two areB gene deletion replicates, one on glucose and one on glycerol were discarded due to bad quality and therefore not included in the analysis. 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.
2011-01-31 | GSE21015 | GEO
Project description:RacA is the main Rho GTPase in Aspergillus niger regulating polarity maintenance via controlling actin dynamics. We have previously shown that both deletion and dominant activation of RacA (RacG18V) provoke an actin localization defect and thereby loss of polarized tip extension. This loss of apical dominance results in frequent dichotomous branching in the racA deletion strain and an apolar growing phenotype for RacG18V. In the current study we investigated the transcriptomics and physiological consequences of these morphological changes and compared the data with our previously established morphogenetic network model for the dichotomous branching mutant ramosa-1. This integrated approach revealed that polar tip growth is most likely orchestrated by the concerted activities of phospholipid signaling, sphingolipid signaling, TORC2 signaling, calcium signaling and CWI signaling pathways. The transcriptomic signatures and the reconstructed network model for all three morphology mutants imply that these pathways become integrated to bring about different physiological adaptations including changes in sterol, zinc and amino acid metabolism and changes in ion transport and protein trafficking. We furthermore followed that fate of exocytotic (SncA) and endocytotic (AbpA, SlaB) markers in the dichotomous branching racA deletion mutant, and provide data demonstrating that hyperbranching does not per se result in increased protein secretion. This data set consists of 14 samples in total and covers the genome-wide transcriptional responses of Aspergillus niger towards deletion of racA and expression of a dominant active racA allele (G18V). The genome-wide transcriptional effect of deleting racA was monitored by comparison with the A. niger wild type. For both, the racA deletion mutant and the wild type, samples consist of biological triplicates (6 in total). The genome-wide transcriptional effect of expressing the dominant active racA mutant allele was monitored by sampling 2 and 4 hours after induction with maltose. For comparison, the racA wild type allele was induced and expressed similarly and samples were taken at corresponding time points. The experimental setup for the dominant active racA expression consisted of biological triplicates (8 in total).
2013-08-25 | E-GEOD-42258 | biostudies-arrayexpress
Project description:Colonies of Aspergillus niger secrete proteins throughout the colony except for the sporulating zone. Inactivation of flbA results in colonies that are unable to reproduce asexually and that secrete proteins throughout the mycelium. In addition, the ΔflbA strain shows cell lysis and has thinner cell walls. This pleiotropic phenotype is associated with differential expression of 38 transcription factor genes. Here, the impact of inactivation of one of these regulatory genes, fusR, was assessed. Mycelium of ΔfusR and its control strain MA234.1 was subjected to whole genome expression analysis, which revealed that 10 out of 15 genes of the fumonisin biosynthetic gene cluster were strongly downregulated in ΔfusR.
2018-02-21 | GSE93990 | GEO
Project description:To evaluate the relative contribution of individual sugar catabolic pathways to the overall physiology of A. niger during growth on plant biomass, the A. niger sugar catabolic pathways involved in converting the most common monomers of plant biomass polysaccharides were blocked, and the phenotype as well as the transcriptomic response of these single-pathway deletion mutants was analysed on two distinct plant biomass substrates: wheat bran (WB) and sugar beet pulp (SBP). Deletion of all the above catabolic pathways were also combined in a single strain (?all), to evaluate whether other compounds of plant biomass could still support growth of A. niger when utilization of the major sugar components is blocked. On both WB and SBP, the strongest affected single-pathway deletion mutants were the PCP and glycolysis deficient mutants, delta-xkiA and delta-hxkA/delta-glkA, respectively. Especially on WB, which is a particularly rich substrate in pentose sugars, blocking PCP caused a significant effect in delta-xkiA with respect to both growth and transcriptomic response. However, delta-hxkA/delta-glkA was shown to be the most critically affected strain. On both substrates, combined deletion of hxkA and glkA resulted in the strongest growth reduction of all the single-pathway mutants, very similar to the multi-pathway deletion mutant delta-all. This result clearly pointed to glycolysis as the most essential and indispensable pathway for the survival of A. niger on these substrates. Since cellulose is not a preferred carbon source for A. niger, it seems that the double deletion of hxkA and glkA mostly affected the utilization of starch. (Note: A portion of this research was performed on a project award (10.46936/expl.proj.2019.51072/60006675) from the Environmental Molecular Sciences Laboratory, a DOE Office of Science User Facility sponsored by the Biological and Environmental Research program under Contract No. DE-AC05-76RL01830)
2022-08-31 | GSE196397 | GEO