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The effects of ClrB internal mutant overexpression and creA deletion on the transcriptome of Penicillium oxalicum

ABSTRACT: The synthesis of cellulolytic enzymes by filamentous fungi are mainly regulated at the transcriptional level. The essential role of transcriptional activator ClrB/CLR-2 in the induction of cellulase genes has been reported in several fungal species. In this study, we identified the C-terminal region of ClrB in Penicillium oxalicum as a transcriptional activation domain using a yeast reporter gene system. Overexpression of a mutant ClrBID fusing the DNA-binding and transcriptional activation domains of ClrB together enabled cellulase production under cellulase non-inducing conditions. Strikingly, the ClrBID-overexpression strain produced cellulase in the presence of glucose at a level similar to that of reference strain on cellulose. Combination of ClrBID overexpression and deletion of carbon catabolite-repressor gene creA suggested an additive effect of the two manipulations on carbon catabolite de-repression. A similar modification of ClrB in Aspergillus niger also resulted in cellulase production on glucose. The results indicated that the post-transcriptional control of ClrB activity involves its central region, and provided a potential strategy for engineering Penicillium and Aspergillus strains for cellulase production on easy-to-use carbon sources.

ORGANISM(S): Penicillium oxalicum

PROVIDER: GSE120416 | GEO | 2019/01/01

REPOSITORIES: GEO

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Inducer Free Cellulase Secretion in Neurospora Crassa and comparitive analysis of cellulase induction in Aspergillus nidulans

Project description:Purpose: To explore conservation of gene regulation by the transcription factor clr-2/clrB in Neurospora crassa and Aspergillus nidulans Methods: mRNA from wild type and clr-2/clrB mutants were collected after a culture shift from sucrose/glucose to Avicel (crystaline cellulose) or no carbon media Results: We show that N. crassa and A. nidulans have similair global transcriptional responses to Avicel, with several hundred genes showing specific induction, though the induced genes are more specifically targeted at cellulose for N. crassa and more targeted at hemicellulose and pectin for A. nidulans. clr-2/clrB has a conserved fundamental function in cellulose induction, though the mechanism has diverged. Misexpression of clr-2 is sufficeint for inducer free cellulase secretion in N. crassa, but neither clrB or heterologous clr-2 is sufficient for inducer free cellulase secretion in A. nidulans. Conclusions: Our study demonstrates a conserved and essential role in cellulose utilization for the transcription factor clr-2 in filamentous ascomycetes and demonstrates that manipulation of clr-2 expression can be used to control cellulase expression in some species.

2013-09-01 | GSE44100 | GEO

Inducer Free Cellulase Secretion in Neurospora Crassa and comparitive analysis of cellulase induction in Aspergillus nidulans

2013-09-01 | E-GEOD-44100 | biostudies-arrayexpress

The effects of transcriptional activator engineering on the transcriptome of Penicillium oxalicum

Project description:The filamentous fungus Penicillium oxalicum can secret various enzymes for efficient saccharification of plant biomass materials. Expression of the constitutively active forms of transcriptional activators ClrB, XlnR and AraR could trigger the production of different sets of lignocellulolytic enzymes. Here, the transcriptomes of the three engineered strains were compared with that of wild type in the medium without carbon source.

2020-12-05 | GSE162657 | GEO

Transcriptomic analysis of cellulolytic fungus Penicillium oxalicum and transcription factor mutant strains in response to different carbon sources

2015-05-28 | GSE69298 | GEO

Transcriptomic analysis of cellulolytic fungus Penicillium oxalicum and transcription factor mutant strains in response to different carbon sources

Project description:RNA-seq was used to compare the responses of Penicillium oxalicum strains to different carbon sources including carbon-starvation, glucose and cellulose. The wild-type strain and transcription facor (ClrB, CreA and AmyR) mutants were studied. A total of 24 samples were analzyed, which include biological triplicates. The wild-type strain cultured without carbon source was considered as a reference.

2015-05-28 | E-GEOD-69298 | biostudies-arrayexpress

The fungus Aspergillus niger consumes sugars in a sequential manner that is not mediated by the carbon catabolite repressor CreA

Project description:Aspergillus niger is a filamentous ascomycete fungus that is commonly found in most biotopes around the globe. In nature, A. niger degrades the plant biomass polysaccharides to monomeric sugars, transports them into the cells, and uses a variety of catabolic pathways to convert them into biochemical building blocks and energy. We show that when grown in liquid cultures, A. niger takes up plant-biomass derived monomeric sugars (and maltose) in a highly sequential manner, rather than simultaneously. Interestingly, this sequential uptake was not mediated by the fungal general carbon catabolite repressor protein CreA, which has been shown to mediate the use of preferential carbon sources over non-preferential carbon sources. Furthermore, transcriptome analysis strongly indicated that the preferential use of the monomeric sugars is arranged at the level of transport, but it is not reflected in transcriptional regulation of sugar catabolism. Therefore, the results indicate that the regulation of sugar transport and catabolism are separate physiological processes in A. niger.

2018-04-23 | GSE98434 | GEO

VIB1, a link between glucose signaling and carbon catabolite repression, is essential for plant cell wall degradation by Neurospora crassa

Project description:Purpose: To explore the function of VIB1 in regulating cellulase production in Neurospora crassa. Method: mRNA from vib-1 mutants were collected after a culture shift from sucrose to Avicel (crystaline cellulose) or carbon-free media. Expression profiles of vib-1 mutants were compared with published profiles of wild type created under the same conditions. Results: We found that many genes that specifically upregulated in wild type upon exposure to Avicel were expressed at low levels in ∆vib-1 and many other genes involved in metabolism and energy were expressed at high levels compared to wild type. Conclusions: Our study shows that VIB1 is required for suppression of glucose response and carbon catabolite repression to allow proper expression of clr-2 and subsequent cellulase production in response to cellulosic induction.

2014-07-08 | GSE52316 | GEO

Studies on the production of fungal polyketides in Aspergillus nidulans using systems biology tools

Project description:Using genetic engineering tools available for the model organism Aspergillus nidulans, we constructed two recombinant strains; one expressing the model polyketide Penicillium griseofulvum 6-methylsalicylic acid (6-MSA) polyketide synthase gene, and one expressing the 6-MSA gene and overexpressing the native phosphoketolase (phk) for increasing the pool of polyketide precursor levels. The physiology of the recombinant strains and a reference wild type were characterized on glucose, xylose, glycerol and ethanol medium in controlled bioreactors. Glucose was found to be the preferable carbon source for 6-MSA production and 6-MSA titers up to 455 mg/L were achieved. Our findings indicate that overexpression of phk does not directly improve 6-MSA production on glucose but if the lower glycolysis is lowered, it is possible to obtain quite high conversion yields of sugar to 6-MSA. Systems biology tools were employed for in-depth analysis of the metabolic processes. Transcriptome analysis of 6-MSA producing strains on glucose and xylose in the presence and absence of phk overexpression combined with flux and physiology data enabled us to propose a model of phk/6msas interaction describing two different responses influencing 6-MSA production. Four strains on two carbon sources

2009-02-27 | E-GEOD-12859 | biostudies-arrayexpress

Genome-wide RNA-seq analysis during carbon catabolite repression in Trichoderma reesei reveals a multilevel control of cellulase production and action

Project description:The ascomycete Trichoderma reesei is one of the most well studied cellulolytic fungi and widely used in the biotechnology industry, as in the production of second-generation bioethanol. Carbon catabolite repression (CCR) mechanism adopted by T. reesei is mediated by the transcription factor CRE1 and consists in the repression of genes related to the production of cellulase when a readily available carbon source is present in the medium. Using RNA sequencing this study aims to contribute to understanding of CCR during growth in cellulose and glucose, by comparing the mutant strain of T. reesei Δcre1 with its parental, QM9414.

2017-05-01 | GSE57374 | GEO

VIB1, a link between glucose signaling and carbon catabolite repression, is essential for plant cell wall degradation by Neurospora crassa

Project description:Purpose: To explore the function of VIB1 in regulating cellulase production in Neurospora crassa. Method: mRNA from vib-1 mutants were collected after a culture shift from sucrose to Avicel (crystaline cellulose) or carbon-free media. Expression profiles of vib-1 mutants were compared with published profiles of wild type created under the same conditions. Results: We found that many genes that specifically upregulated in wild type upon exposure to Avicel were expressed at low levels in M-bM-^HM-^Fvib-1 and many other genes involved in metabolism and energy were expressed at high levels compared to wild type. Conclusions: Our study shows that VIB1 is required for suppression of glucose response and carbon catabolite repression to allow proper expression of clr-2 and subsequent cellulase production in response to cellulosic induction. Biological triplicates of liquid culture N. crassa were harvested at 4 hours after a switch from sucrose media to media of interest. Global mRNA abundances were measured by sequencing on the Illumina Genome Analyzer Iix and HiSeq2000 platforms.

2014-07-08 | E-GEOD-52316 | biostudies-arrayexpress

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