scholarly journals Broad Substrate-Specific Phosphorylation Events Are Associated With the Initial Stage of Plant Cell Wall Recognition in Neurospora crassa

2019 ◽  
Vol 10 ◽  
Author(s):  
Maria Augusta C. Horta ◽  
Nils Thieme ◽  
Yuqian Gao ◽  
Kristin E. Burnum-Johnson ◽  
Carrie D. Nicora ◽  
...  
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Initial Stage

scholarly journals Direct Target Network of the Neurospora crassa Plant Cell Wall Deconstruction Regulators CLR-1, CLR-2, and XLR-1

mBio ◽  
2015 ◽  
Vol 6 (5) ◽  
Author(s):  
James P. Craig ◽  
Samuel T. Coradetti ◽  
Trevor L. Starr ◽  
N. Louise Glass
Keyword(s):  
Gene Expression ◽  
Cell Wall ◽  
Transcription Factors ◽  
Neurospora Crassa ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Nutrient Sensing ◽  
Wall Material ◽  
Content Type ◽  
Genes Encoding

ABSTRACTFungal deconstruction of the plant cell requires a complex orchestration of a wide array of intracellular and extracellular enzymes. InNeurospora crassa, CLR-1, CLR-2, and XLR-1 have been identified as key transcription factors regulating plant cell wall degradation in response to soluble sugars. The XLR-1 regulon was defined using a constitutively active mutant allele, resulting in hemicellulase gene expression and secretion under noninducing conditions. To define genes directly regulated by CLR-1, CLR-2, and XLR-1, we performed chromatin immunoprecipitation and next-generation sequencing (ChIPseq) on epitope-tagged constructs of these three transcription factors. WhenN. crassais exposed to plant cell wall material, CLR-1, CLR-2, and XLR-1 individually bind to the promoters of the most strongly induced genes in their respective regulons. These include promoters of genes encoding cellulases for CLR-1 and CLR-2 (CLR-1/CLR-2) and promoters of genes encoding hemicellulases for XLR-1. CLR-1 bound to its regulon under noninducing conditions; however, this binding alone did not translate into gene expression and enzyme secretion. Motif analysis of the bound genes revealed conserved DNA binding motifs, with the CLR-2 motif matching that of its closest paralog inSaccharomyces cerevisiae, Gal4p. Coimmunoprecipitation studies showed that CLR-1 and CLR-2 act in a homocomplex but not as a CLR-1/CLR-2 heterocomplex.IMPORTANCEUnderstanding fungal regulation of complex plant cell wall deconstruction pathways in response to multiple environmental signals via interconnected transcriptional circuits provides insight into fungus/plant interactions and eukaryotic nutrient sensing. Coordinated optimization of these regulatory networks is likely required for optimal microbial enzyme production.

scholarly journals Systems analysis of plant cell wall degradation by the model filamentous fungus Neurospora crassa

2009 ◽  
Vol 106 (52) ◽  
pp. 22157-22162 ◽  
Author(s):  
Chaoguang Tian ◽  
William T. Beeson ◽  
Anthony T. Iavarone ◽  
Jianping Sun ◽  
Michael A. Marletta ◽  
...  
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Plant Cell ◽  
Filamentous Fungus ◽  
Systems Analysis ◽  
Plant Cell Wall ◽  
Cell Wall Degradation ◽  
Plant Cell Wall Degradation

scholarly journals Network reconstruction and systems analysis of plant cell wall deconstruction by Neurospora crassa

2017 ◽  
Vol 10 (1) ◽  
Author(s):  
Areejit Samal ◽  
James P. Craig ◽  
Samuel T. Coradetti ◽  
J. Philipp Benz ◽  
James A. Eddy ◽  
...  
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Plant Cell ◽  
Systems Analysis ◽  
Plant Cell Wall ◽  
Network Reconstruction

scholarly journals Identification of Glutaminyl Cyclase Genes Involved in Pyroglutamate Modification of Fungal Lignocellulolytic Enzymes

mBio ◽  
2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Vincent W. Wu ◽  
Craig M. Dana ◽  
Anthony T. Iavarone ◽  
Douglas S. Clark ◽  
N. Louise Glass
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Filamentous Fungi ◽  
Plant Cell ◽  
Mammalian Cells ◽  
Thermal Denaturation ◽  
Plant Cell Wall ◽  
Industrial Plant ◽  
Amino Acid Derivative ◽  
Lignocellulolytic Enzymes

ABSTRACT The breakdown of plant biomass to simple sugars is essential for the production of second-generation biofuels and high-value bioproducts. Currently, enzymes produced from filamentous fungi are used for deconstructing plant cell wall polysaccharides into fermentable sugars for biorefinery applications. A post-translational N-terminal pyroglutamate modification observed in some of these enzymes occurs when N-terminal glutamine or glutamate is cyclized to form a five-membered ring. This modification has been shown to confer resistance to thermal denaturation for CBH-1 and EG-1 cellulases. In mammalian cells, the formation of pyroglutamate is catalyzed by glutaminyl cyclases. Using the model filamentous fungus Neurospora crassa , we identified two genes ( qc - 1 and qc - 2 ) that encode proteins homologous to mammalian glutaminyl cyclases. We show that qc - 1 and qc - 2 are essential for catalyzing the formation of an N-terminal pyroglutamate on CBH-1 and GH5-1. CBH-1 and GH5-1 produced in a Δqc - 1 Δqc - 2 mutant, and thus lacking the N-terminal pyroglutamate modification, showed greater sensitivity to thermal denaturation, and for GH5-1, susceptibility to proteolytic cleavage. QC-1 and QC-2 are endoplasmic reticulum (ER)-localized proteins. The pyroglutamate modification is predicted to occur in a number of additional fungal proteins that have diverse functions. The identification of glutaminyl cyclases in fungi may have implications for production of lignocellulolytic enzymes, heterologous expression, and biotechnological applications revolving around protein stability. IMPORTANCE Pyroglutamate modification is the post-translational conversion of N-terminal glutamine or glutamate into a cyclized amino acid derivative. This modification is well studied in animal systems but poorly explored in fungal systems. In Neurospora crassa , we show that this modification takes place in the ER and is catalyzed by two well-conserved enzymes, ubiquitously conserved throughout the fungal kingdom. We demonstrate that the modification is important for the structural stability and aminopeptidase resistance of CBH-1 and GH5-1, two important cellulase enzymes utilized in industrial plant cell wall deconstruction. Many additional fungal proteins predicted in the genome of N. crassa and other filamentous fungi are predicted to carry an N-terminal pyroglutamate modification. Pyroglutamate addition may also be a useful way to stabilize secreted proteins and peptides, which can be easily produced in fungal production systems.

Production of plant cell wall degrading enzymes by the exo-1 mutant and wild type Neurospora crassa

2010 ◽  
Vol 150 ◽  
pp. 513-514
Author(s):  
A.M. Polizeli ◽  
M.A. Moraes ◽  
J.A. Jorge ◽  
H.F. Terenzi ◽  
M.L.T.M. Polizeli
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Wild Type ◽  
Cell Wall Degrading Enzymes ◽  
Degrading Enzymes

scholarly journals Broad substrate-specific phosphorylation events are associated with the initial stage of plant cell wall recognition in Neurospora crassa

2019 ◽  
Author(s):  
Maria Augusta Crivelente Horta ◽  
Nils Thieme ◽  
Yuqian Gao ◽  
Kristin E. Burnum-Johnson ◽  
Carrie D. Nicora ◽  
...  
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Plant Cell ◽  
Large Scale ◽  
Map Kinases ◽  
Molecular Mechanisms ◽  
Plant Cell Wall ◽  
High Specificity ◽  
Cell Wall Degradation ◽  
Plant Cell Wall Degradation

AbstractFungal plant cell wall degradation processes are governed by complex regulatory mechanisms, allowing the organisms to adapt their metabolic program with high specificity to the available substrates. While the uptake of representative plant cell wall mono- and disaccharides is known to induce specific transcriptional and translational responses, the processes related to early signal reception and transduction remain largely unkown. A fast and reversible way of signal transmission are post-translational protein modifications, such as phosphorylations, which could initiate rapid adaptations of the fungal metabolism to a new condition. To elucidate how changes in the initial substrate recognition phase of Neurospora crassa affect the global phosphorylation pattern, phospho-proteomics was performed after a short (2 minutes) induction period with several plant cell wall-related mono- and disaccharides. The MS/MS-based peptide analysis revealed large-scale substrate-specific protein phosphorylation and de-phosphorylations. Using the proteins identified by MS/MS, a protein-protein-interaction (PPI) network was constructed. The variance in phosphorylation of a large number of kinases, phosphatases and transcription factors indicate the participation of many known signaling pathways, including circadian responses, two-component regulatory systems, MAP kinases as well as the cAMP-dependent and heterotrimeric G-protein pathways. Adenylate cyclase, a key component of the cAMP pathway, was identified as a potential hub for carbon source-specific differential protein interactions. In addition, four phosphorylated F-Box proteins were identified, two of which, Fbx-19 and Fbx-22, were found to be involved in carbon catabolite repression responses. Overall, these results provide unprecedented and detailed insights into a so far less well known stage of the fungal response to environmental cues and allow to better elucidate the molecular mechanisms of sensory perception and signal transduction during plant cell wall degradation.

scholarly journals Network reconstruction and systems analysis of plant cell wall deconstruction byneurospora crassa

2017 ◽  
Author(s):  
Areejit Samal ◽  
James P. Craig ◽  
Samuel T. Coradetti ◽  
J. Philipp Benz ◽  
James A. Eddy ◽  
...  
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Filamentous Fungi ◽  
Plant Cell ◽  
Plant Cell Wall ◽  
Plant Biomass ◽  
Degradation Pathway ◽  
Heterogeneous Data ◽  
Data Types ◽  
Cell Wall Polysaccharides

AbstractPlant biomass degradation by fungal derived enzymes is rapidly expanding in economic importance as a clean and efficient source for biofuels. The ability to rationally engineer filamentous fungi would facilitate biotechnological applications for degradation of plant cell wall polysaccharides. However, incomplete knowledge of biomolecular networks responsible for plant cell wall deconstruction impedes experimental efforts in this direction. To expand this knowledge base, a detailed network of reactions important for deconstruction of plant cell wall polysaccharides into simple sugars was constructed for the filamentous fungusNeurospora crassa. To reconstruct this network, information was integrated from five heterogeneous data types: functional genomics, transcriptomics, proteomics, genetics, and biochemical characterizations. The combined information was encapsulated into a feature matrix and the evidence weighed to assign annotation confidence scores for each gene within the network. Comparative analyses of RNA-seq and ChIP-seq data shed light on the regulation of the plant cell wall degradation network (PCWDN), leading to a novel hypothesis for degradation of the hemicellulose mannan. The transcription factor CLR-2 was subsequently experimentally shown to play a key role in the mannan degradation pathway ofNeurospora crassa. Our network serves as a scaffold for integration of diverse experimental data, leading to elucidation of regulatory design principles for plant cell wall deconstruction by filamentous fungi, and guiding efforts to rationally engineer industrially relevant hyper-production strains.

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