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Deletion of the FlbA-regulated transcription factor msnB of Aspergillus niger affects protein secretion

ABSTRACT: 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.

ORGANISM(S): Aspergillus niger

PROVIDER: GSE104669 | GEO | 2018/10/01

REPOSITORIES: GEO

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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.

2018-06-01 | GSE102899 | GEO

The FlbA-regulated transcription factor FusR of Aspergillus niger is involved in production of fumonisin

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

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

Project description: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.

2014-02-13 | GSE44391 | GEO

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

Project description:Degradation of plant biomass to fermentable sugars is of critical importance for the use of plant materials for biofuels. Filamentous fungi are ubiquitous organisms and major plant biomass degraders. Single colonies of some fungal species can colonize massive areas as large as five soccer stadia.During growth, the mycelium encounters heterogeneous carbon sources. Here we assessed whether substrate heterogeneity is a major determinant of spatial gene expression in colonies of Aspergillus niger. We analyzed whole-genome gene expression in five concentric zones of 5-day-old colonies utilizing sugar beet pulp as a complex carbon source. Growth, protein production and secretion occurred throughout the colony. Genes involved in carbon catabolism and nitrate utilization were expressed uniformly from the centre to the periphery whereas genes encoding plant biomass degrading enzymes were expressed differentially across the colony. A combined adaptive response of carbon-catabolism and enzyme production to locally available monosaccharides was observed. Finally, our results demonstrate a fine-tuning of the different enzymatic tools available in A. niger and reveal how this fungus adapts as it colonizes complex environments.

2015-09-09 | PXD002130 | Pride

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

Project description:Filamentous fungi are ubiquitous organisms and major plant biomass degraders. As a single colony, some fungal species can colonize large areas as up to five soccer stadia. During growth, the mycelium encounters heterogeneous carbon sources. Here we assessed whether substrate heterogeneity is a main determinant of spatial gene expression in colonies of Aspergillus niger. This question was addressed by analyzing whole-genome gene expression in five concentric zones of 5-day-old sugar beet pulp grown colonies. Growth, protein production and secretion were occurring throughout the whole colony. Carbon and nitrate utilization were constant from the centre to the periphery whereas genes encoding plant cell wall degrading enzymes were expressed with a different pattern across the colony. Finally our results demonstrate a fine-tuning of the different enzymatic tools available in A. niger and expand the knowledge on how this fungus adapts as it colonizes complex environments.

2016-03-04 | GSE66641 | GEO

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.

2014-02-13 | E-GEOD-44391 | biostudies-arrayexpress

Transcription profiling of Bacillus cereus variants that are filametous or short-celled in the exponential phase

Project description:While some members of the Bacillus cereus group constitutively display filamentous growth, other members of this group may grow filamentously as a result of environmental stimuli or genetic changes. In this study, two variants of B. cereus ATCC 14579 displaying filamentous growth during exponential phase were compared with a variant which grew as single cells or short chains during this stage of growth. Phenotypic differences between the variants were investigated by a range of methods, comparing the variants at the genomic, transcriptional and translational level to search for changes which could explain the observed phenotypes. Differences were found between filamentous and purely planktonic variants as well as between the filamentous variants. Filamentous variants displayed rougher colony morphology on agar plates, and did not lyse after death or sporulation. One filamentous variant additionally showed reduced sporulation and increased transcription of class I and and III heat shock proteins, but these phenotypes were not found in the other filamentous variant.

2010-07-01 | E-TABM-924 | biostudies-arrayexpress

Regulating the angiogenic balance in tissues

Project description:A balance between angiogenesis inducers and inhibitors in the microenvironment controls the rate of new blood vessel formation. We hypothesized that fibroblasts, an important cellular constituent of the tissue stroma, secrete molecules that contribute to this balance. We further hypothesized that fibroblasts secrete molecules that promote angiogenesis when they are in a proliferative state and molecules that inhibit angiogenesis when they are not actively cycling (quiescent). Microarray analysis revealed that angiogenesis inducers and inhibitors are regulated as fibroblasts transition into a quiescent state and reenter the cell cycle in response to changes in serum. To assess whether changes in transcript levels result in changes in the levels of secreted proteins, we collected conditioned medium from proliferating and quiescent fibroblasts and performed immunoblotting for selected proteins. Secreted protein levels of the angiogenesis inhibitor pigment epithelium derived factor (PEDF) were higher in quiescent than proliferating fibroblasts. Conversely, proliferating fibroblasts secreted increased levels of the angiogenesis inducer vascular endothelial growth factor-C (VEGF-C). For the angiogenesis inhibitor thrombospondin-2, quiescent cells secreted a prominent 160 kDa form in addition to the 200 kDa form secreted by proliferating and restimulated fibroblasts. Using immunohistochemistry we discovered that fibroblasts surround blood vessels and that the angiogenesis inhibitor PEDF is expressed by quiescent fibroblasts in uterine tissue, supporting a role for PEDF in maintaining quiescence of the vasculature. This work takes a new approach to the study of angiogenesis by examining the expression of multiple angiogenesis regulators secreted from a key stromal cell, the fibroblast. mRNAs were analyzed by two color microarray from a human neonatal dermal fibroblasts cell line over a timecourse of serum starvation and serum restimulation.

2012-12-03 | E-GEOD-42681 | biostudies-arrayexpress

Expression data from Aspergillus niger comparing aerial structures with vegetative mycelium

Project description:A. niger undergoes dramatic changes during asexual development. We tried to identify the differences in RNA expression levels that are important for this development. We used micro-arrays to determine which genes were up- or down-regulated in the aerial structures, the part of the colony that is formed during asexual development. A. niger was grown as a sandwiched culture (Wösten et al., 1991,Journal of General Microbiology 137: 2017–2023) in a 0.25 mm layer of 0.6 % agarose between two porous polycarbonate membranes (diameter 76 mm, pore size 0.1 µm; Profiltra; www.profiltra.nl). After six days of growth, the top membrane of the sandwich was replaced by a membrane with pores of 10 µm (Profiltra), allowing formation of aerial hyphae and conidiophores for 24 h. Vegetative mycelium and aerial structures of 7-day-old maltose-grown cultures of A. niger were harvested from 3 and 5 sandwiched colonies, respectively. The aerial structures were scraped from the top membrane of the sandwiched culture with a razor blade. From other colonies, vegetative mycelium was harvested by flipping over the top membrane and scraping it off with a razor blade.

2012-02-24 | E-GEOD-32123 | biostudies-arrayexpress

Aspergillus niger WT: Control vs. steam-exploded sugarcane induction

Project description:Transcriptional profiling of A. niger comparing WT strain vs. ΔXlnR strain treated with steam-exploded sugarcane bagasse (SESB) for 6, 12 and 24 h. The main objective was to identifiy genes related to cellulases and hemicellulases, comparing the differences between WT strain and the strain with the disrupted xylanolytic transcriptional activator gene, XlnR, after treatment with steam-exploded sugarcane. The experiment was further validated by real-time PCR, mass spectrometry of secreted proteins and enzymatic assays.

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