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

ABSTRACT: 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.

ORGANISM(S): Neurospora crassa

PROVIDER: GSE52316 | GEO | 2014/07/08

SECONDARY ACCESSION(S): PRJNA227384

REPOSITORIES: GEO

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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

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

Conserved and Essential Transcription Factors for Cellulase Gene Expression in Ascomycete Fungi.

Project description:Transcriptional profiling with next-generation sequencing methods refined our understanding of the N. crassa transcriptional response to cellulose and demonstrated that the newly characterized transcription factors clr-1 and clr-2 were required for the bulk of that response including induction all major cellulase and some major hemicellulase genes. N. crassa pregrown in Sucrose and transferred to Avicel (cellulose), Sucrose or media with no carbon added. Biological triplicates used to identify differentially expressed genes in WT. Single libraries for mutant strains identify which genes show deficient regulation in response to Avicel. Note: Samples named "cdr1" and "cdr2" correspond to the genes clr-1 and clr-2 respectively.

2012-04-23 | E-GEOD-35227 | biostudies-arrayexpress

QUANTITATIVE PROTEOME PROFILING OF NEUROSPORA CRASSA REVEALS A CELLOBIOSE-DEPENDENT NETWORK COORDINATING LIGNOCELLULOLYTIC RESPONSES

Project description:Cellulose, particularly the major cellulolytic product cellobiose, can induce the production of enzymes associated with deconstruction of lignocellulose in filamentous fungi. However, the detailed mechanisms underlying this biotechnologically important process remain to be disclosed. Here, the proteome response to cellobiose, crystalline cellulose (Avicel), and carbon starvation of a Neurospora crassa triple β-glucosidase mutant were compared using tandem mass tag (TMT)-based proteome quantification. Improved quantification accuracy was achieved with synchronous precursor selection (SPS)-based MS3 technology compared to MS2 using a high resolution tribrid mass spectrometer. Exposure to carbon starvation, cellobiose or Avicel induced the production of cellulase and lytic polysaccharide monooxygenase enzymes in N. crassa, as well as a cellobionic acid transporter, indicating their functional roles in the early adaptation to plant cell wall. In particular, cellobiose specifically induced the production of proteins in the functional categories of protein processing and export as well as cell wall organization. The data presented here integrates the signaling pathway associated with cellobiose transporters CDT-1 and/or CDT-2 with the direct targets of the transcription factors CLR-1, CLR-2, and XLR-1, the unfolded protein response (UPR) mediated by Ire-1/Hac-1, as well as calcium homeostasis and cell wall organization. The cellobiose-dependent response network will be useful for rational strain improvement to facilitate the production of lignocellulases in filamentous fungi and plant biomass-based products.

2020-05-15 | PXD011598 | Pride

A novel conserved transcription factor CLR-4 regulates cellulase genes expression through controlling the intracellular cAMP level in ascomycete fungi

Project description:Fungal degradation of lignocellulosic biomass requires various (hemi-)cellulases and plays key roles in biological carbon cycle. Although cellulases induction recently described in some saprobic filamentous fungi, regulation of cellulase transcription has not been studied thoroughly. Here, we identified and characterized the novel cellulase regulation factors clr-4 in Neurospora crassa and its ortholog Mtclr-4 in Myceliophthora thermophila. Deletion of clr-4 and Mtclr-4 displayed similarly defective phenotypes in cellulolytic enzymes production and activities. Transcriptomics analysis of Δclr-4/ΔMtclr-4 revealed down-regulation of not only encoding genes of (hemi-)cellulases and pivotal regulators (clr-1, clr-2 and xyr-1), but also the key genes of cAMP signaling pathway such as adenylate cyclase cr-1. Consistently, the significant decreased levels of intracellular cAMP were observed in Δclr-4/ΔMtclr-4 compared to wild-type during cellulose utilization. Electrophoretic mobility shift assays (EMSA) verified that CLR-4 could directly bind to the promoter regions of adenylyl cyclase (Nccr-1) and cellulose regulator clr-1, while MtCLR-4 bind to upstream regions of adenylyl cyclase Mtcr-1 and biomass deconstruction regulators Mtclr-2 and Mtxyr-1. Concluded, the novel cellulase expression regulators (CLR-4/MtCLR-4) findings here significantly enrich our understanding of the regulatory network of cellulose degradation and provide new targets for industrial fungi strain engineering for plant biomass deconstruction in biorefinery.

2021-03-18 | GSE111986 | GEO

Evidence of a critical role for cellodextrin transporter 2 (CDT-2) in both cellulose and hemicellulose degradation and utilization in Neurospora crassa

Project description:CDT-1 and CDT-2 are two cellodextrin transporters discovered in the filamentous fungus Neurospora crassa. Previous studies focused on characterizing the role of these transporters in only a few conditions, including cellulose degradation, and the function of these two transporters is not yet completely understood. In this study, we show that deletion of cdt-2, but not cdt-1, results in growth defects not only on Avicel but also on xylan. cdt-2 can be highly induced by xylan, and this mutant has a xylodextrin consumption defect. Transcriptomic analysis of the cdt-2 deletion strain on Avicel and xylan showed that major cellulase and hemicellulase genes were significantly down-regulated in the cdt-2 deletion strain and artificial over expression of cdt-2 in N. crassa increased cellulase and hemicellulase production. Together, these data clearly show that CDT-2 plays a critical role in hemicellulose sensing and utilization. This is the first time a sugar transporter has been assigned a function in the hemicellulose degradation pathway. Furthermore, we found that the transcription factor XLR-1 is the major regulator of cdt-2, while cdt-1 is primarily regulated by CLR-1. These results deepen our understanding of the functions of both cellodextrin transporters, particularly for CDT-2. Our study also provides novel insight into the mechanisms for hemicellulose sensing and utilization in N. crassa, and may be applicable to other cellulolytic filamentous fungi. N. crassa was pregrown in Sucrose and transferred to Avicel (cellulose) or Xylan(hemicellulose) media. Up regulated and down regulated genes expressions were compared with wild type strain on two conditions (Avicel and xylan) respectively.

2014-09-25 | E-GEOD-44673 | biostudies-arrayexpress

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

Project description: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.

2019-01-01 | GSE120416 | GEO

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

Transcriptome study of the deletion of vacuolar protein sorting related vps13 and vps21 in Trichoderma reesei

Project description:The disruption of vacuolar protein sorting related genes vps13 and vps21 in filamentou fungi Trichoderma reesei resulted in a dramtic increase in cellulase transcription especially in the late Avicel induction phase, relieving the RESS (REpression under Secretion Stress) effect. Great interest arisen concered about the reason and underlying mechanism of vps13 and vps21 disruption for increased cellulase transcription. Thus, transcriptome analysis through RNA-Seq could give a deep insight of the potential pathways involved in enhanced cellulase transcription in late Avicel induction.

2021-11-10 | GSE187573 | GEO

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