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The putative cellodextrin transporter-like protein CLP1 is involved in cellulase induction in Neurospora crassa

ABSTRACT: Neurospora crassa recently has become a novel system to investigate cellulase induction. Here, we discovered a novel membrane protein, CLP1 (NCU05853), a putative cellodextrin transporter-like protein, that is a critical component of the cellulase induction pathway in N. crassa. Although CLP1 protein cannot transport cellodextrin, the suppression of cellulase induction by this protein was discovered on both cellobiose and Avicel. The co-disruption of the cellodextrin transporters cdt2 and clp1 in strain Δ3βG formed strain CPL7. With induction by cellobiose, cellulase production was enhanced 6.9-fold in CPL7 compared with Δ3βG. We also showed that the suppression of cellulase expression by CLP1 occurred by repressing the expression of cellodextrin transporters, particularly cdt1 expression. Transcriptome analysis of the hypercellulase-producing strain CPL7 showed that the cellulase expression machinery was dramatically stimulated, as were the cellulase enzyme genes including the inducer transporters and the major transcriptional regulators.

ORGANISM(S): Neurospora crassa OR74A

PROVIDER: GSE60004 | GEO | 2014/11/13

SECONDARY ACCESSION(S): PRJNA257279

REPOSITORIES: GEO

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The putative cellodextrin transporter-like protein CLP1 is involved in cellulase induction in Neurospora crassa

Project description:Neurospora crassa recently has become a novel system to investigate cellulase induction. Here, we discovered a novel membrane protein, CLP1 (NCU05853), a putative cellodextrin transporter-like protein, that is a critical component of the cellulase induction pathway in N. crassa. Although CLP1 protein cannot transport cellodextrin, the suppression of cellulase induction by this protein was discovered on both cellobiose and Avicel. The co-disruption of the cellodextrin transporters cdt2 and clp1 in strain M-NM-^T3M-NM-2G formed strain CPL7. With induction by cellobiose, cellulase production was enhanced 6.9-fold in CPL7 compared with M-NM-^T3M-NM-2G. We also showed that the suppression of cellulase expression by CLP1 occurred by repressing the expression of cellodextrin transporters, particularly cdt1 expression. Transcriptome analysis of the hypercellulase-producing strain CPL7 showed that the cellulase expression machinery was dramatically stimulated, as were the cellulase enzyme genes including the inducer transporters and the major transcriptional regulators. N. crassa was pregrown in sucrose and transferred to cellobiose media. Up regulated and down regulated genes expressions were compared with M-NM-^T3M-NM-2G and M-NM-^T3M-NM-2GM-NM-^Tclp1 strain.

2014-11-13 | E-GEOD-60004 | biostudies-arrayexpress

The Protein Non-anchored cell wall protein NCW-1 promotes cellulase production through effects on cellobiose uptake in Neurospora crassa

Project description:The mechanism of cellulase induction in the filamentous fungi is still not completely understood, and the components of this pathway are not well characterized. In this study, the mechanism of how the non-anchored cell wall protein NCW-1 (NCU05137) affect the cellulases induction was investigated. Transcriptome analysis of this quadruple deletion strain △3βG△ncw-1 showed that the expression of major cellulase, cellodextrin transporters and cellulase regulators were significantly increased. Taken together, the cellulase expression machinery was dramatically stimulated in Δ3βGΔncw-1. These results make us understanding the ncw-1 function better and deepened our knowledge about the cellulase induction in N. crassa.

2016-09-30 | GSE73838 | 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

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

2014-09-25 | GSE44673 | GEO

Neurospora crassa OR74A

Project description:The putative cellodextrin transporter-like protein CLP1 is involved in cellulase induction in Neurospora crassa

| PRJNA257279 | ENA

Impact of plant cell wall derived sugars on transcriptional and post-transcriptional control in Saccharomyces cerevisiae

Project description:Saccharomyces cerevisiae cannot metabolize non-glucose sugars including cellobiose, xylose, xylodextrins in nature, which are prevalent in plant cell wall. Here, one engineered S. cerevisiae strain, which expresses a cellodextrin transporter gene (cdt-1) and an intracellular β-glucosidase gene (codon-optimized gh1-1) from Neurospora crassa; XYL1 (xylose reductase gene), XYL2 (xylitol dehydrogenase gene), and XKS1 (xylulose kinase gene) from Scheffersomyces stipitis, as well as cdt-2 (coding for cellodextrin transporter 2), gh43-2 (coding for β-xylosidase) and gh43-7 (coding for a xylosyl-xylitol-specific β-xylosidase) from N. crassa, can utilize the above non-glucose sugars. We sequenced mRNA from exponential cultures of the engineered S. cerevisiae grown on glucose, cellobiose, xylose or xylodextrins as a single carbon source in both aerobic and anaerobic conditions in biological triplicate. Differences in gene expression between non-glucose sugar and glucose metabolism revealed by RNA deep sequencing indicated that non-glucose sugar metabolism induced mitochondrial activation and reduced amino acid and protein biosynthesis under fermentation conditions.

2021-05-27 | GSE69404 | GEO

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

Transcriptional reprogramming of engineered cellobiose-utilizing Saccharomyces cerevisiae in response to cellobiose revealed by RNA-Seq

2015-04-10 | GSE54825 | GEO

Transcriptional reprogramming of engineered cellobiose-utilizing Saccharomyces cerevisiae in response to cellobiose revealed by RNA-Seq

Project description:Saccharomyces cerevisiae cannot metabolize cellobiose in nature. Here, S. cerevisiae was engineered to achieve cellobiose utilization by introducing both a cellodextrin transporter gene (cdt-1) and an intracellular β-glucosidase gene (gh1-1) from Neurospora crassa. We sequenced mRNA from anaerobic exponential cultures of engineered S. cerevisiae grown on cellobiose or glucose as a single carbon source in biological triplicate. Differences in gene expression between cellobiose and glucose metabolism revealed by RNA deep sequencing indicated that cellobiose metabolism induced mitochondrial activation and reduced amino acid biosynthesis under fermentation conditions. mRNA levels in cellobiose-grown and glucose-grown cells of engineered cellobiose-utilizing Saccharomyces cerevisiae were examined by deep sequencing, in triplicate, using Illumina Genome Analyzer-II. We sequenced 3 samples from cellobiose-grown cells and 3 samples from glucose-grown cells and identified differential expressions in the cellobiose versus glucose fermentations by using mRNA levels of glucose-grown cells as a reference.

2015-04-10 | E-GEOD-54825 | biostudies-arrayexpress

Effect of the res-2 transcription factor gene deletion on protein secretion and stress response in the hyperproducer strain Trichoderma reesei Rut-C30

Project description:The fungus Trichoderma reesei is one of the most used industrial cellulase producers due to its high capacity of protein secretion. The strain was shown to possess an expanded endoplasmic reticulum, but the genetic factors responsible for this phenotype remain still unidentified. Recently, three new transcription factors were described in Neurospora crassa which were demonstrated to be involved in protein secretion. The orthologous gene of one of them, Res-2, was deleted in the T. reesei hyperproducer strain Rut-C30 and its role on protein secretion analyzed. The mutant strain showed slower growth on all substrates tested, but the presence of the stress agent DTT had contrasting effects. Protein secretion tended to be lower in the deletion strain compared to the parental strain Rut-C30. Analysis of the transcriptomes of the Rut-C30 and the Δres-2 mutant strain in secretion stress conditions reveiled remarkably few differences, but induction of cellulase secretion and/or presence of DTT led to differential expression of genes coding for transporters, redox genes and lipid metabolism. These results suggest that in the T. reesei Rut-C30 strain, Res-2 does not act as a master regulator of the secretion pathway, but contributes to a high protein secretion by adjusting the expression of genes involved in membrane turnover and the redox status of the organism or specific organelles.

2023-11-14 | GSE233738 | GEO

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