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specificity and regulation of peptide transport on neurospora crassa. 2002128318
localization of enzymes in the cell walls of neurospora crassa. 2002127745
effects of inhibitors on the plasma membrane and mitochondrial adenosine triphosphatases of neurospora crassa.a comparative study has been made of the effects of a variety of inhibitors on the plasma membrane atpase and mitochondrial atpase of neurospora crassa. the most specific inhibitors proved to be vanadate and diethylstilbestrol for the plasma membrane atpase and azide, oligomycin, venturicidin, and leucinostatin for mitochondrial atpase. n,n'-dicyclohexylcarbodiimide, octylguanidine, triphenylsulfonium chloride, and quercetin and related bioflavonoids inhibited both enzymes, although with differe ...2003151557
fast light-regulated genes of neurospora crassa.several physiological reactions including the sexual differentiation of the ascomycete neurospora crassa are triggered by blue light. mutants in the white-collar genes wc-1 and wc-2 are blind for all the blue light effects tested so far. we have previously shown that blue light induces some translatable mrnas at different times after beginning the illumination. here we report the cdna cloning of four genes that are induced by blue light. induction of these transcripts is temporally ordered (lag ...20082527354
proteases of neurospora crassa: their role in morphology. 2002127746
gene topography and function. i. gene expression in germinating conidia of neurospora crassa.in an attempt to find clues for the significance of the gene ordering along the eukaryotic chromosome, a system consisting of germinating conida of neurospora crassa was studied. thirteen enzyme activities corresponding to genes widely distributed on five chromosomes were determined in dormant and in germinating conidia. ten of these enzymes showed lower activities in the resting state, and the time patterns of their increase were determined during germination. the results obtained do not suppor ...2002128350
characterization of neurospora crassa polyadenylated messenger ribonucleic acid: structure of the 5'-terminus. 2003148868
neurospora crassa temperature-sensitive mutant apparently defective in protein synthesis.a temperature-sensitive mutant of neurospora was isolated which appeared to be defective in the initiation of protein synthesis. the defect in mutant 34cts was apparently due to a single gene mutation, and was recessive in heterokaryons. conidial germination was normal and hyphal growth was nearly so in the mutant at 20 c, but both were greatly inhibited at 35 c. after 15 min at 35 c there was a reduced rate of protein synthesis, followed by decreases in ribonucleic acid and deoxyribonucleic aci ...2002123244
spin-labeled mitochondrial lipids in neurospora crassa. 20144301148
the structure of an rnai polymerase links rna silencing and transcription.rna silencing refers to a group of rna-induced gene-silencing mechanisms that developed early in the eukaryotic lineage, probably for defence against pathogens and regulation of gene expression. in plants, protozoa, fungi, and nematodes, but apparently not insects and vertebrates, it involves a cell-encoded rna-dependent rna polymerase (crdrp) that produces double-stranded rna triggers from aberrant single-stranded rna. we report the 2.3-a resolution crystal structure of qde-1, a crdrp from neur ...200617147473
a widespread class of reverse transcriptase-related cellular genes.reverse transcriptases (rts) polymerize dna on rna templates. they fall into several structurally related but distinct classes and form an assemblage of rt-like enzymes that, in addition to rts, also includes certain viral rna-dependent rna polymerases (rdrp) synthesizing rna on rna templates. it is generally believed that most rt-like enzymes originate from retrotransposons or viruses and have no specific function in the host cell, with telomerases being the only notable exception. here we repo ...201121876125
identification and biochemical characterization of molybdenum cofactor-binding proteins from arabidopsis thaliana.the molybdenum cofactor (moco) forms part of the catalytic center in all eukaryotic molybdenum enzymes and is synthesized in a highly conserved pathway. among eukaryotes, very little is known about the processes taking place subsequent to moco biosynthesis, i.e. moco transfer, allocation, and insertion into molybdenum enzymes. in the model plant arabidopsis thaliana, we identified a novel protein family consisting of nine members that after recombinant expression are able to bind moco with k(d) ...200920040598
crystal structure and ligand binding of the mid domain of a eukaryotic argonaute protein.argonaute (ago) proteins are core components of rna-induced silencing complexes and have essential roles in rna-mediated gene silencing. they are characterized by a bilobal architecture, consisting of one lobe containing the amino-terminal and paz domains and another containing the mid and piwi domains. except for the paz domain, structural information on eukaryotic ago domains is not yet available. in this study, we report the crystal structure of the mid domain of the eukaryotic ago protein qd ...201020539312
crystal structure of the mid-piwi lobe of a eukaryotic argonaute protein.argonaute proteins (agos) are essential effectors in rna-mediated gene silencing pathways. they are characterized by a bilobal architecture, in which one lobe contains the n-terminal and paz domains and the other contains the mid and piwi domains. here, we present the first crystal structure of the mid-piwi lobe from a eukaryotic ago, the neurospora crassa qde-2 protein. compared to prokaryotic agos, the domain orientation is conserved, indicating a conserved mode of nucleic acid binding. the pi ...201121646546
the uve1 endonuclease is regulated by the white collar complex to protect cryptococcus neoformans from uv damage.the pathogenic fungus cryptococcus neoformans uses the bwc1-bwc2 photoreceptor complex to regulate mating in response to light, virulence and ultraviolet radiation tolerance. how the complex controls these functions is unclear. here, we identify and characterize a gene in cryptococcus, uve1, whose mutation leads to a uv hypersensitive phenotype. the homologous gene in fission yeast schizosaccharomyces pombe encodes an apurinic/apyrimidinic endonuclease acting in the uvde-dependent excision repai ...201324039606
physiological role of acyl coenzyme a synthetase homologs in lipid metabolism in neurospora crassa.acyl coenzyme a (coa) synthetase (acs) enzymes catalyze the activation of free fatty acids (fas) to coa esters by a two-step thioesterification reaction. activated fas participate in a variety of anabolic and catabolic lipid metabolic pathways, including de novo complex lipid biosynthesis, fa β-oxidation, and lipid membrane remodeling. analysis of the genome sequence of the filamentous fungus neurospora crassa identified seven putative fatty acss (acs-1 through acs-7). acs-3 was found to be the ...201323873861
parallel structural evolution of mitochondrial ribosomes and oxphos complexes.the five macromolecular complexes that jointly mediate oxidative phosphorylation (oxphos) in mitochondria consist of many more subunits than those of bacteria, yet, it remains unclear by which evolutionary mechanism(s) these novel subunits were recruited. even less well understood is the structural evolution of mitochondrial ribosomes (mitoribosomes): while it was long thought that their exceptionally high protein content would physically compensate for their uniquely low amount of ribosomal rna ...201525861818
characterization of cercospora nicotianae hypothetical proteins in cercosporin resistance.the photoactivated toxin, cercosporin, produced by cercospora species, plays an important role in pathogenesis of this fungus to host plants. cercosporin has almost universal toxicity to cells due to its production of reactive oxygen species including singlet oxygen. for that reason, cercospora species, which are highly resistant to their own toxin, are good candidates to identify genes for resistance to cercosporin and to the reactive oxygen species it produces. in previous research, the zinc c ...201526474162
mutagenesis of trichoderma reesei endoglucanase i: impact of expression host on activity and stability at elevated temperatures.trichoderma reesei is a key cellulase source for economically saccharifying cellulosic biomass for the production of biofuels. lignocellulose hydrolysis at temperatures above the optimum temperature of t. reesei cellulases (~50°c) could provide many significant advantages, including reduced viscosity at high-solids loadings, lower risk of microbial contamination during saccharification, greater compatibility with high-temperature biomass pretreatment, and faster rates of hydrolysis. these potent ...201525879765
molecular evolution and functional characterisation of an ancient phenylalanine ammonia-lyase gene (nnpal1) from nelumbo nucifera: novel insight into the evolution of the pal family in angiosperms.phenylalanine ammonia-lyase (pal; e.c.4.3.1.5) is a key enzyme of the phenylpropanoid pathway in plant development, and it catalyses the deamination of phenylalanine to trans-cinnamic acid, leading to the production of secondary metabolites. this enzyme has been identified in many organisms, ranging from prokaryotes to higher plants. because nelumbo nucifera is a basal dicot rich in many secondary metabolites, it is a suitable candidate for research on the phenylpropanoid pathway.201424884360
stress-induced cell death is mediated by ceramide synthesis in neurospora crassa.the combined stresses of moderate heat shock (45 degrees c) and analog-induced glucose deprivation constitute a lethal stress for neurospora crassa. we found that this cell death requires fatty acid synthesis and the cofactor biotin. in the absence of the cofactor, the stressed cells are particularly sensitive to exogenous ceramide, which is lethal at low concentrations. when we extracted endogenous sphingolipids, we found that unique ceramides were induced (i) by the inhibitory glucose analog 2 ...200818952903
czt-1 is a novel transcription factor controlling cell death and natural drug resistance in neurospora crassa.we pinpoint czt-1 (cell death-activated zinc cluster transcription factor) as a novel transcription factor involved in tolerance to cell death induced by the protein kinase inhibitor staurosporine in neurospora crassa. transcriptional profiling of staurosporine-treated wild-type cells by rna-sequencing showed that genes encoding the machinery for protein synthesis are enriched among the genes repressed by the drug. functional category enrichment analyses also show that genes encoding components ...201424717808
a circadian oscillator in the fungus botrytis cinerea regulates virulence when infecting arabidopsis thaliana.the circadian clock of the plant model arabidopsis thaliana modulates defense mechanisms impacting plant-pathogen interactions. nevertheless, the effect of clock regulation on pathogenic traits has not been explored in detail. moreover, molecular description of clocks in pathogenic fungi--or fungi in general other than the model ascomycete neurospora crassa--has been neglected, leaving this type of question largely unaddressed. we sought to characterize, therefore, the circadian system of the pl ...201526124115
light activation of the lov protein vivid generates a rapidly exchanging dimer.the fungal photoreceptor vivid (vvd) plays an important role in the adaptation of blue-light responses in neurospora crassa. vvd, an fad-binding lov (light, oxygen, voltage) protein, couples light-induced cysteinyl adduct formation at the flavin ring to conformational changes in the n-terminal cap (ncap) of the vvd pas domain. size-exclusion chromatography (sec), equilibrium ultracentrifugation, and static and dynamic light scattering show that these conformational changes generate a rapidly exc ...200818553928
illuminating solution responses of a lov domain protein with photocoupled small-angle x-ray scattering.the pas-lov domain is a signal-transducing component found in a large variety of proteins that is responsible for sensing different stimuli such as light, oxygen, and voltage. the lov protein vvd regulates blue light responses in the filamentous fungi neurospora crassa. using photocoupled, time-resolved small-angle x-ray scattering, we extract the solution protein structure in both dark-adapted and light-activated states. two distinct dark-adapted conformations are detected in the wild-type prot ...200919712683
the l-cysteine desulfurase nfs1 is localized in the cytosol where it provides the sulfur for molybdenum cofactor biosynthesis in humans.in humans, the l-cysteine desulfurase nfs1 plays a crucial role in the mitochondrial iron-sulfur cluster biosynthesis and in the thiomodification of mitochondrial and cytosolic trnas. we have previously demonstrated that purified nfs1 is able to transfer sulfur to the c-terminal domain of mocs3, a cytosolic protein involved in molybdenum cofactor biosynthesis and trna thiolation. however, no direct evidence existed so far for the interaction of nfs1 and mocs3 in the cytosol of human cells. here, ...201323593335
phosphate starvation in fungi induces the replacement of phosphatidylcholine with the phosphorus-free betaine lipid diacylglyceryl-n,n,n-trimethylhomoserine.diacylglyceryl-n,n,n-trimethylhomoserine (dgts) is a phosphorus-free betaine-lipid analog of phosphatidylcholine (ptdcho) synthesized by many soil bacteria, algae, and nonvascular plants. synthesis of dgts and other phosphorus-free lipids in bacteria occurs in response to phosphorus (p) deprivation and results in the replacement of phospholipids by nonphosphorous lipids. the genes encoding dgts biosynthetic enzymes have previously been identified and characterized in bacteria and the alga chlamy ...201424728191
systems biology of the qa gene cluster in neurospora crassa.an ensemble of genetic networks that describe how the model fungal system, neurospora crassa, utilizes quinic acid (qa) as a sole carbon source has been identified previously. a genetic network for qa metabolism involves the genes, qa-1f and qa-1s, that encode a transcriptional activator and repressor, respectively and structural genes, qa-2, qa-3, qa-4, qa-x, and qa-y. by a series of 4 separate and independent, model-guided, microarray experiments a total of 50 genes are identified as qa-respon ...201121695121
array comparative genomic hybridizations: assessing the ability to recapture evolutionary relationships using an in silico approach.comparative genomic hybridization (cgh) with dna microarrays has many biological applications including surveys of copy number changes in tumorogenesis, species detection and identification, and functional genomics studies among related organisms. array cgh has also been used to infer phylogenetic relatedness among species or strains. although the use of the entire genome can be seen as a considerable advantage for use in phylogenetic analysis, few such studies have questioned the reliability of ...201121936922
characterization of the insertase for β-barrel proteins of the outer mitochondrial membrane.the tob-sam complex is an essential component of the mitochondrial outer membrane that mediates the insertion of β-barrel precursor proteins into the membrane. we report here its isolation and determine its size, composition, and structural organization. the complex from neurospora crassa was composed of tob55-sam50, tob38-sam35, and tob37-sam37 in a stoichiometry of 1:1:1 and had a molecular mass of 140 kd. a very minor fraction of the purified complex was associated with one mdm10 protein. usi ...201223128244
the oxygenase cao-1 of neurospora crassa is a resveratrol cleavage enzyme.the genome of the ascomycete neurospora crassa encodes cao-1 and cao-2, two members of the carotenoid cleavage oxygenase family that target double bonds in different substrates. previous studies demonstrated the role of cao-2 in cleaving the c40 carotene torulene, a key step in the synthesis of the c35 apocarotenoid pigment neurosporaxanthin. in this work, we investigated the activity of cao-1, assuming that it may provide retinal, the chromophore of the nop-1 rhodopsin, by cleaving β-carotene. ...201323893079
potential conservation of circadian clock proteins in the phylum nematoda as revealed by bioinformatic searches.although several circadian rhythms have been described in c. elegans, its molecular clock remains elusive. in this work we employed a novel bioinformatic approach, applying probabilistic methodologies, to search for circadian clock proteins of several of the best studied circadian model organisms of different taxa (mus musculus, drosophila melanogaster, neurospora crassa, arabidopsis thaliana and synechoccocus elongatus) in the proteomes of c. elegans and other members of the phylum nematoda. wi ...201425396739
molecular tools for functional genomics in filamentous fungi: recent advances and new strategies.advances in genetic transformation techniques have made important contributions to molecular genetics. various molecular tools and strategies have been developed for functional genomic analysis of filamentous fungi since the first dna transformation was successfully achieved in neurospora crassa in 1973. increasing amounts of genomic data regarding filamentous fungi are continuously reported and large-scale functional studies have become common in a wide range of fungal species. in this review, ...201323988676
establishment of crispr/cas9 in alternaria alternata.the filamentous fungus alternaria alternata is a potent producer of many secondary metabolites, some of which like alternariol or alternariol-methyl ether are toxic and/or cancerogenic. many alternaria species do not only cause post-harvest losses of food and feed, but are aggressive plant pathogens. despite the great economic importance and the large number of research groups working with the fungus, the molecular toolbox is rather underdeveloped. gene deletions often result in heterokaryotic s ...201728286319
role of the alternaria alternata blue-light receptor lrea (white-collar 1) in spore formation and secondary metabolism.alternaria alternata is a filamentous fungus that causes considerable loss of crops of economically important feed and food worldwide. it produces more than 60 different secondary metabolites, among which alternariol (aoh) and altertoxin (atx) are the most important mycotoxins. we found that mycotoxin production and spore formation are regulated by light in opposite ways. whereas spore formation was largely decreased under light conditions, the production of aoh was stimulated 2- to 3-fold. atx ...201424532063
microbial transformations of isophorone by alternaria alternata and neurospora crassa.isophorone (3,5,5-trimethyl-2-cyclohexen-1-one), a monoterpene, and the structurally related 1,8-cineole and camphor, have demonstrated a protective effect against cancer, biological activity against a variety of microorganisms, and anti-oxidant properties. the derivatization of isophorone is, therefore, an important field of xenobiochemistry, pharmacology and toxicology. the aim of this study was to obtain derivatives of isophorone through microbial biotransformation and evaluate the biotransfo ...201323472460
the woronin body in the nematophagous fungus arthrobotrys oligospora is essential for trap formation and efficient pathogenesis.the woronin body is a unique organelle in pezizomycotina species. following the injury of hyphae, it can quickly seal the septal pores to reduce the loss of cytoplasm and promote hyphal healing. the woronin body is also considered a significant factor in efficient pathogenesis in many fungal pathogens. in this study, we identified aohex1, a homologue of neurospora crassa hex1, a main component of the woronin body in the genome of the nematode-trapping fungus arthrobotrys oligospora. to study the ...201728007213
genomic characteristics comparisons of 12 food-related filamentous fungi in trna gene set, codon usage and amino acid composition.filamentous fungi are widely exploited in food industry due to their abilities to secrete large amounts of enzymes and metabolites. the recent availability of fungal genome sequences has provided an opportunity to explore the genomic characteristics of these food-related filamentous fungi. in this paper, we selected 12 representative filamentous fungi in the areas of food processing and safety, which were aspergillus clavatus, a. flavus, a. fumigatus, a. nidulans, a. niger, a. oryzae, a. terreus ...201222305983
gliotoxin effects on fungal growth: mechanisms and exploitation.although initially investigated for its antifungal properties, little is actually known about the effect of gliotoxin on aspergillus fumigatus and other fungi. we have observed that exposure of a. fumigatus to exogenous gliotoxin (14 μg/ml), under gliotoxin-limited growth conditions, results in significant alteration of the expression of 27 proteins (up- and down-regulated >1.9-fold; p<0.05) including de novo expression of cu, zn superoxide dismutase, up-regulated allergen asp f3 expression and ...201222405895
fungal sex: the ascomycota.this article provides an overview of sexual reproduction in the ascomycetes, a phylum of fungi that is named after the specialized sacs or "asci" that hold the sexual spores. they have therefore also been referred to as the sac fungi due to these characteristic structures that typically contain four to eight ascospores. ascomycetes are morphologically diverse and include single-celled yeasts, filamentous fungi, and more complex cup fungi. the sexual cycles of many species, including those of the ...201627763253
rbdb, a rhomboid protease critical for srebp activation and virulence in aspergillus fumigatus.srebp transcription factors play a critical role in fungal virulence; however, the mechanisms of sterol regulatory element binding protein (srebp) activation in pathogenic fungi remains ill-defined. screening of the neurospora crassa whole-genome deletion collection for genes involved in hypoxia responses identified a gene for an uncharacterized rhomboid protease homolog, rbdb, required for growth under hypoxic conditions. loss of rbdb in aspergillus fumigatus also inhibited growth under hypoxic ...201727303716
identification and characterization of a novel aspergillus fumigatus rhomboid family putative protease, rbda, involved in hypoxia sensing and virulence.aspergillus fumigatus is the most common pathogenic mold infecting humans and a significant cause of morbidity and mortality in immunocompromised patients. in invasive pulmonary aspergillosis, a. fumigatus spores are inhaled into the lungs, undergoing germination and invasive hyphal growth. the fungus occludes and disrupts the blood vessels, leading to hypoxia and eventual tissue necrosis. the ability of this mold to adapt to hypoxia is regulated in part by the sterol regulatory element binding ...201627068092
structural divergence of the group i intron binding surface in fungal mitochondrial tyrosyl-trna synthetases that function in rna splicing.the mitochondrial tyrosyl-trna synthetases (mttyrrss) of pezizomycotina fungi, a subphylum that includes many pathogenic species, are bifunctional proteins that both charge mitochondrial trna(tyr) and act as splicing cofactors for autocatalytic group i introns. previous studies showed that one of these proteins, neurospora crassa cyt-18, binds group i introns by using both its n-terminal catalytic and c-terminal anticodon binding domains and that the catalytic domain uses a newly evolved group i ...201627036943
functional characterization of the woronin body protein wsca of the pathogenic mold aspergillus fumigatus.woronin bodies are fungal-specific organelles that seal damaged hyphal compartments and thereby contribute to the stress resistance and virulence of filamentous fungi. in this study, we have characterized the aspergillus fumigatus woronin body protein wsca. wsca is homologous to neurospora crassa wsc, a protein that was shown to be important for biogenesis, segregation and positioning of woronin bodies. wsca and wsc both belong to the mpv17/pmp22 family of peroxisomal membrane proteins. an a. fu ...201627016805
de-repression of csp-1 activates adaptive responses to antifungal azoles.antifungal azoles are the major drugs that are used to treat fungal infections. this study found that in response to antifungal azole stress, neurospora crassa could activate the transcriptional responses of many genes and increase azole resistance by reducing the level of conidial separation 1 (csp-1), a global transcription repressor, at azole-responsive genes. the expression of csp-1 was directly activated by the transcription factors wc-1 and wc-2. upon ketoconazole (ktc) stress, the transcr ...201626781458
fgric8 is involved in regulating vegetative growth, conidiation, deoxynivalenol production and virulence in fusarium graminearum.proteins of the resistance to inhibitors of cholinesterase 8 (ric8) group act as guanine nucleotide exchange factors (gefs) and play important roles in regulating g-protein signaling in animals. in filamentous fungi, putative ric8 orthologs have so far been identified in magnaporthe oryzae, neurospora crassa, aspergillus nidulans and aspergillus fumigatus. here, we report the functional investigation of a potential ric8 ortholog (fgric8) in the wheat head blight pathogen fusarium graminearum. ta ...201526341536
transcription factor ads-4 regulates adaptive responses and resistance to antifungal azole stress.azoles are commonly used as antifungal drugs or pesticides to control fungal infections in medicine and agriculture. fungi adapt to azole stress by rapidly activating the transcription of a number of genes, and transcriptional increases in some azole-responsive genes can elevate azole resistance. the regulatory mechanisms that control transcriptional responses to azole stress in filamentous fungi are not well understood. this study identified a bzip transcription factor, ads-4 (antifungal drug s ...201526100701
the apses family proteins in fungi: characterizations, evolution and functions.the apses protein family belongs to transcriptional factors of the basic helix-loop-helix (bhlh) class, the originally described members (apses: asm1p, phd1p, sok2p, efg1p and stuap) are used to designate this group of proteins, and they have been identified as key regulators of fungal development and other biological processes. apses proteins share a highly conserved dna-binding domain (apses domain) of about 100 amino acids, whose central domain is predicted to form a typical bhlh structure. b ...201525534868
predicting essential genes for identifying potential drug targets in aspergillus fumigatus.aspergillus fumigatus (af) is a ubiquitous and opportunistic pathogen capable of causing acute, invasive pulmonary disease in susceptible hosts. despite current therapeutic options, mortality associated with invasive af infections remains unacceptably high, increasing 357% since 1980. therefore, there is an urgent need for the development of novel therapeutic strategies, including more efficacious drugs acting on new targets. thus, as noted in a recent review, "the identification of essential ge ...201424569026
phosphorylation of calcineurin at a novel serine-proline rich region orchestrates hyphal growth and virulence in aspergillus fumigatus.the fungus aspergillus fumigatus is a leading infectious killer in immunocompromised patients. calcineurin, a calmodulin (cam)-dependent protein phosphatase comprised of calcineurin a (cnaa) and calcineurin b (cnab) subunits, localizes at the hyphal tips and septa to direct a. fumigatus invasion and virulence. here we identified a novel serine-proline rich region (sprr) located between two conserved cnaa domains, the cnab-binding helix and the cam-binding domain, that is evolutionarily conserved ...201323990785
fungal siderophore biosynthesis is partially localized in peroxisomes.siderophores play a central role in iron metabolism and virulence of most fungi. both aspergillus fumigatus and aspergillus nidulans excrete the siderophore triacetylfusarinine c (tafc) for iron acquisition. in a. fumigatus, green fluorescence protein-tagging revealed peroxisomal localization of the tafc biosynthetic enzymes sidi (mevalonyl-coa ligase), sidh (mevalonyl-coa hydratase) and sidf (anhydromevalonyl-coa transferase), while elimination of the peroxisomal targeting signal (pts) impaired ...201323617799
fungal cell wall organization and biosynthesis.the composition and organization of the cell walls from saccharomyces cerevisiae, candida albicans, aspergillus fumigatus, schizosaccharomyces pombe, neurospora crassa, and cryptococcus neoformans are compared and contrasted. these cell walls contain chitin, chitosan, β-1,3-glucan, β-1,6-glucan, mixed β-1,3-/β-1,4-glucan, α-1,3-glucan, melanin, and glycoproteins as major constituents. a comparison of these cell walls shows that there is a great deal of variability in fungal cell wall composition ...201323419716
two functional motifs define the interaction, internalization and toxicity of the cell-penetrating antifungal peptide paf26 on fungal cells.the synthetic, cell penetrating hexapeptide paf26 (rkkwfw) is antifungal at low micromolar concentrations and has been proposed as a model for cationic, cell-penetrating antifungal peptides. its short amino acid sequence facilitates the analysis of its structure-activity relationships using the fungal models neurospora crassa and saccharomyces cerevisiae, and human and plant pathogens aspergillus fumigatus and penicillium digitatum, respectively. previously, paf26 at low fungicidal concentration ...201323349973
the transcriptional response to the inactivation of the pampk1 and pampk2 map kinase pathways in podospora anserina.transcription pattern during mycelium growth of podospora anserina was assayed by microarray analysis in wild type and in mutants affected in the map kinase genes pampk1 and pampk2 and in the nadph oxidase gene panox1. 15% of the genes have their expression modified by a factor two or more as growth proceeds in wild type. the genes whose expression is modified during growth in p. anserina are either not conserved or differently regulated in neurospora crassa and aspergillus niger, two fungi for ...201222721649
a comparative study of hydrolysis and transglycosylation activities of fungal β-glucosidases.β-glucosidases (bgs) from aspergillus fumigatus, aspergillus niger, aspergillus oryzae, magnaporthe grisea, neurospora crassa, and penicillium brasilianum were purified to homogeneity, and investigated for their (simultaneous) hydrolytic and transglycosylation activity in samples with high concentrations of either cellobiose or glucose. the rate of the hydrolytic process (which converts one cellobiose to two glucose molecules) shows a maximum around 10-15 mm cellobiose and decreases with further ...201322311644
aspergillus glaucus aquaglyceroporin gene glpf confers high osmosis tolerance in heterologous organisms.aquaglyceroporins (glpfs) that transport glycerol along with water and other uncharged solutes are involved in osmoregulation in myriad species. fungal species form a large group of eukaryotic organisms, and their glpfs may be diverse, exhibiting various activities. however, few filamentous fungal glpfs have been biologically investigated. here, a glpf gene from the halophilic fungus aspergillus glaucus (agglpf) was verified to be a channel of water or glycerol in xenopus laevis oocytes and was ...201526209670
improved gene targeting frequency in marine-derived filamentous fungus aspergillus glaucus by disrupting ligd.filamentous fungi from the marine environment have shown great potential as cell factories for the production of pharmacologically active metabolites, but extremely low frequency of homologous recombination brings difficulty to further molecular biology studies. to bypass this problem and develop a highly efficient gene targeting system in marine-derived filamentous fungus aspergillus glaucus, ligd, a homolog of neurospora crassa mus-53 which is considered to play a significant role in nonhomolo ...201222562375
an evolutionarily conserved transcriptional activator-repressor module controls expression of genes for d-galacturonic acid utilization in aspergillus niger.the expression of genes encoding extracellular polymer-degrading enzymes and the metabolic pathways required for carbon utilization in fungi are tightly controlled. the control is mediated by transcription factors that are activated by the presence of specific inducers, which are often monomers or monomeric derivatives of the polymers. a d-galacturonic acid-specific transcription factor named gaar was recently identified and shown to be an activator for the expression of genes involved in galact ...201728049705
molecular components of the neurospora crassa ph signaling pathway and their regulation by ph and the pac-3 transcription factor.environmental ph induces a stress response triggering a signaling pathway whose components have been identified and characterized in several fungi. neurospora crassa shares all six components of the aspergillus nidulans ph signaling pathway, and we investigate here their regulation during an alkaline ph stress response. we show that the n. crassa pal mutant strains, with the exception of δpal-9, which is the a. nidulans pali homolog, exhibit low conidiation and are unable to grow at alkaline ph. ...201627557053
light governs asexual differentiation in the grey mould fungus botrytis cinerea via the putative transcription factor bcltf2.botrytis cinerea is a plant pathogenic fungus known for its utilization of light as environmental cue to regulate asexual differentiation: conidia are formed in the light, while sclerotia are formed in the dark. as no orthologues of known regulators of conidiation (e.g., aspergillus nidulans brla, neurospora crassa fl) exist in the leotiomycetes, we initiated a de novo approach to identify the functional counterpart in b. cinerea. the search revealed the light-responsive c2h2 transcription facto ...201627347834
gln3 is a main regulator of nitrogen assimilation in candida glabrata.after candida albicans, the yeast candida glabrata ranks second as an aetiological agent of candidaemia and is the most frequently encountered non-candida albicans species in patients with invasive candidiasis. transcriptome analysis in c. albicans, c. glabrata and cryptoccocus neoformans has revealed that, when engulfed by macrophages, these yeasts upregulate genes involved in nutrient acquisition, including nitrogen transporters such as the general amino acid permease gap1, the dicarboxylic am ...201627222014
extreme diversity in the regulation of ndt80-like transcription factors in fungi.the saccharomyces cerevisiae ndt80 protein is the founding member of a class of p53-like transcription factors that is known as the ndt80/phog-like dna-binding family. the number of ndt80-like genes in different fungi is highly variable and their roles, which have been examined in only a few species, include regulation of meiosis, sexual development, biofilm formation, drug resistance, virulence, the response to nutrient stress and programmed cell death. the protein kinase ime2 regulates the sin ...201526497142
small heat shock proteins, phylogeny in filamentous fungi and expression analyses in aspergillus nidulans.small heat shock proteins (shsps) have been characterized in organisms from all three domains of life and viruses and are involved in a wide range of biological functions. however, the evolution and function of shsp in aspergillus species are largely unknown. in the present work, shsps were identified in 31 filamentous fungi, including species from aspergillus, penicillium, fusarium and magnaporthe, as well as botrytis cinerea and neurospora crassa. phylogenetic analysis revealed high level of d ...201626403724
seeing the world differently: variability in the photosensory mechanisms of two model fungi.light plays an important role for most organisms on this planet, serving either as a source of energy or information for the adaptation of biological processes to specific times of day. the fungal kingdom is estimated to contain well over a million species, possibly 10-fold more, and it is estimated that a majority of the fungi respond to light, eliciting changes in several physiological characteristics including pathogenesis, development and secondary metabolism. two model organisms for photobi ...201626373782
epigenetics as an emerging tool for improvement of fungal strains used in biotechnology.filamentous fungi are today a major source of industrial biotechnology for the production of primary and secondary metabolites, as well as enzymes and recombinant proteins. all of them have undergone extensive improvement strain programs, initially by classical mutagenesis and later on by genetic manipulation. thereby, strategies to overcome rate-limiting or yield-reducing reactions included manipulating the expression of individual genes, their regulatory genes, and also their function. yet, re ...201526115753
a lipid-managing program maintains a stout spitzenkörper.the spitzenkörper (spk) is an accumulation of vesicles interleaved with actin microfilaments present at the cytosolic side of the apical plasma membrane (pm) of hyphal tips of many species of filamentous fungi. the physiological role of the spk has captivated fungal biologists over the years, but only very recently this 'organelle' is starting to be understood in the molecular terminology used for cell biological models. one aspect that has received little attention is the role of cellular membr ...201525921726
regulation of glycogen metabolism by the cre-1, rco-1 and rcm-1 proteins in neurospora crassa. the role of cre-1 as the central transcriptional regulator.the transcription factor crea/mig1/cre-1 is a repressor protein that regulates the use of alternative carbon sources via a mechanism known as carbon catabolite repression (ccr). in saccharomyces cerevisiae, mig1 recruits the complex ssn6-tup1, the neurospora crassa rcm-1 and rco-1 orthologous proteins, respectively, to bind to promoters of glucose-repressible genes. we have been studying the regulation of glycogen metabolism in n. crassa and the identification of the rco-1 corepressor as a regul ...201525889113
transient transformation of podosphaera xanthii by electroporation of conidia.powdery mildew diseases are a major phytosanitary issue causing important yield and economic losses in agronomic, horticultural and ornamental crops. powdery mildew fungi are obligate biotrophic parasites unable to grow on culture media, a fact that has significantly limited their genetic manipulation. in this work, we report a protocol based on the electroporation of fungal conidia, for the transient transformation of podosphaera fusca (synonym podosphaera xanthii), the main causal agent of cuc ...201525651833
dissecting the function of the different chitin synthases in vegetative growth and sexual development in neurospora crassa.chitin, one of the most important carbohydrates of the fungal cell wall, is synthesized by chitin synthases (chs). seven sequences encoding chss have been identified in the genome of neurospora crassa. previously, chs-1, -3 and -6 were found at the spitzenkörper(spk) core and developing septa. we investigated the functional importance of each chs in growth and development of n. crassa. the cellular distribution of each chs tagged with fluorescent proteins and the impact of corresponding gene del ...201525596036
neurospora crassa asm-1 complements the conidiation defect in a stua mutant of aspergillus nidulans.aspergillus nidulans stua and neurospora crassa asm-1 are orthologous apses (asm-1, phd1, sok2, efg1, stua) transcription factors conserved across a diverse group of fungi. stua and asm-1 have roles in asexual (conidiation) and sexual (ascospore formation) development in both organisms. to address the hypothesis that the last common ancestor of these diverse fungi regulated conidiation with similar genes, asm-1 was introduced into the stua1 mutant of a. nidulans. expression of asm-1 complemented ...201525550299
nitrogen regulation of fungal secondary metabolism in fungi.fungi occupy diverse environments where they are constantly challenged by stressors such as extreme ph, temperature, uv exposure, and nutrient deprivation. nitrogen is an essential requirement for growth, and the ability to metabolize a wide variety of nitrogen sources enables fungi to colonize different environmental niches and survive nutrient limitations. favored nitrogen sources, particularly ammonium and glutamine, are used preferentially, while the expression of genes required for the use ...201425506342
kinesin-3 in the basidiomycete ustilago maydis transports organelles along the entire microtubule array.the molecular motor kinesin-3 transports early endosomes along microtubules in filamentous fungi. it was reported that kinesin-3 from the ascomycete fungi aspergillus nidulans and neurospora crassa use a subset of post-translationally modified and more stable microtubules. here, i show that kinesin-3 from the basidiomycete ustilago maydis moves along all hyphal microtubules. this difference is likely due to variation in cell cycle control and associated organization of the microtubule array.201525459534
the aspergillus nidulans zn(ii)2cys6 transcription factor an5673/rhar mediates l-rhamnose utilization and the production of α-l-rhamnosidases.various plant-derived substrates contain l-rhamnose that can be assimilated by many fungi and its liberation is catalyzed by α-l-rhamnosidases. initial data obtained in our laboratory focussing on two aspergillus nidulans α-l-rhamnosidase genes (rhaa and rhae) showed α-l-rhamnosidase production to be tightly controlled at the level of transcription by the carbon source available. whilst induction is effected by l-rhamnose, unlike many other glycosyl hydrolase genes repression by glucose and othe ...201425416526
neurospora crassa female development requires the pacc and other signal transduction pathways, transcription factors, chromatin remodeling, cell-to-cell fusion, and autophagy.using a screening protocol we have identified 68 genes that are required for female development in the filamentous fungus neurospora crassa. we find that we can divide these genes into five general groups: 1) genes encoding components of the pacc signal transduction pathway, 2) other signal transduction pathway genes, including genes from the three n. crassa map kinase pathways, 3) transcriptional factor genes, 4) autophagy genes, and 5) other miscellaneous genes. complementation and rip studies ...201425333968
bem46 shows eisosomal localization and association with tryptophan-derived auxin pathway in neurospora crassa.bem46 proteins are evolutionarily conserved, but their functions remain elusive. we reported previously that the bem46 protein in neurospora crassa is targeted to the endoplasmic reticulum (er) and is essential for ascospore germination. in the present study, we established a bem46 knockout strain of n. crassa. this δbem46 mutant exhibited a level of ascospore germination lower than that of the wild type but much higher than those of the previously characterized bem46-overexpressing and rna inte ...201424928924
interdependence of the actin and the microtubule cytoskeleton during fungal growth.cell polarization is a theme in biology conserved from bacteria to man. one of the most extremely polarized cells in nature is the hyphae of filamentous fungi. a continuous flow of secretion vesicles from the hyphal cell body to the tip is essential for cell wall and membrane extension. microtubules (mts) and actin, along with their corresponding motor proteins, are involved in the secretion process. therefore, the arrangement of the cytoskeleton is a crucial step to establish and maintain polar ...201424879477
[mechanisms and regulation of enzymatic hydrolysis of cellulose in filamentous fungi: classical cases and new models].cellulose is the most abundant renewable carbon source on earth. however, this polymer structure comprises a physical and chemical barrier for carbon access, which has limited its exploitation. in nature, only a few percentage of microorganisms may degrade this polymer by cellulase expression. filamentous fungi are one of the most active and efficient groups among these microorganisms. this review describes similarities and differences between cellulase activity mechanisms and regulatory mechani ...201524607657
genetic characterization of the neurospora crassa molybdenum cofactor biosynthesis.molybdenum (mo) is a trace element that is essential for important cellular processes. to gain biological activity, mo must be complexed in the molybdenum cofactor (moco), a pterin derivative of low molecular weight. moco synthesis is a multi-step pathway that involves a variable number of genes in eukaryotes, which are assigned to four steps of eukaryotic moco biosynthesis. moco biosynthesis mutants lack any moco-dependent enzymatic activities, including assimilation of nitrate (plants and fung ...201424569084
the transcription factor bcltf1 regulates virulence and light responses in the necrotrophic plant pathogen botrytis cinerea.botrytis cinerea is the causal agent of gray mold diseases in a range of dicotyledonous plant species. the fungus can reproduce asexually by forming macroconidia for dispersal and sclerotia for survival; the latter also participate in sexual reproduction by bearing the apothecia after fertilization by microconidia. light induces the differentiation of conidia and apothecia, while sclerotia are exclusively formed in the absence of light. the relevance of light for virulence of the fungus is not o ...201424415947
assessing the effects of light on differentiation and virulence of the plant pathogen botrytis cinerea: characterization of the white collar complex.organisms are exposed to a tough environment, where acute daily challenges, like light, can strongly affect several aspects of an individual's physiology, including pathogenesis. while several fungal models have been widely employed to understand the physiological and molecular events associated with light perception, various other agricultural-relevant fungi still remain, in terms of their responsiveness to light, in the dark. the fungus botrytis cinerea is an aggressive pathogen able to cause ...201324391918
proteomic alterations induced by ionic liquids in aspergillus nidulans and neurospora crassa.this study constitutes the first attempt to understand at the proteomic level the fungal response to ionic liquid stress. ascomycota are able to grow in media supplemented with high concentrations of an ionic liquid, which, in turn, lead to major alterations in the fungal metabolic footprint. herein, we analysed the differential accumulation of mycelial proteins in aspergillus nidulans and neurospora crassa after their exposure to two of the most commonly used ionic liquids: 1-ethyl-3-methylimid ...201324120530
analysis of a conserved cellulase transcriptional regulator reveals inducer-independent production of cellulolytic enzymes in neurospora crassa.cellulose is recalcitrant to deconstruction to glucose for use in fermentation strategies for biofuels and chemicals derived from lignocellulose. in neurospora crassa, the transcriptional regulator, clr-2, is required for cellulolytic gene expression and cellulose deconstruction. to assess conservation and divergence of cellulase gene regulation between fungi from different ecological niches, we compared clr-2 function with its ortholog (clrb) in the distantly related species, aspergillus nidula ...201323766336
neurospora crassa nkin2, a kinesin-3 motor, transports early endosomes and is required for polarized growth.biological motors are molecular nanomachines, which convert chemical energy into mechanical forces. the combination of mechanoenzymes with structural components, such as the cytoskeleton, enables eukaryotic cells to overcome entropy, generate molecular gradients, and establish polarity. hyphae of filamentous fungi are among the most polarized cells, and polarity defects are most obvious. here, we studied the role of the kinesin-3 motor, nkin2, in neurospora crassa. we found that nkin2 localizes ...201323687116
conf and conj contribute to conidia germination and stress response in the filamentous fungus aspergillus nidulans.light induces various responses in fungi including formation of asexual and sexual reproductive structures. the formation of conidia in the filamentous fungus aspergillus nidulans is regulated by red and blue light receptors. expression of conidia associated con genes, which are widely spread in the fungal kingdom, increases upon exposure to light. we have characterized the light-inducible conf and conj genes of a. nidulans which are homologs of con-6 and con-10 of neurospora crassa. con genes a ...201323644150
genetic control of asexual sporulation in filamentous fungi.asexual sporulation (conidiation) in the ascomycetous filamentous fungi involves the formation of conidia, formed on specialized structures called conidiophores. conidiation in filamentous fungi involves many common themes including spatial and temporal regulation of gene expression, specialized cellular differentiation, intra-/inter-cellular communications, and response to environmental factors. the commencement, progression and completion of conidiation are regulated by multiple positive and n ...201223092920
high abundance of serine/threonine-rich regions predicted to be hyper-o-glycosylated in the secretory proteins coded by eight fungal genomes.o-glycosylation of secretory proteins has been found to be an important factor in fungal biology and virulence. it consists in the addition of short glycosidic chains to ser or thr residues in the protein backbone via o-glycosidic bonds. secretory proteins in fungi frequently display ser/thr rich regions that could be sites of extensive o-glycosylation. we have analyzed in silico the complete sets of putatively secretory proteins coded by eight fungal genomes (botrytis cinerea, magnaporthe grise ...201222994653
ambient ph controls glycogen levels by regulating glycogen synthase gene expression in neurospora crassa. new insights into the ph signaling pathway.glycogen is a polysaccharide widely distributed in microorganisms and animal cells and its metabolism is under intricate regulation. its accumulation in a specific situation results from the balance between glycogen synthase and glycogen phosphorylase activities that control synthesis and degradation, respectively. these enzymes are highly regulated at transcriptional and post-translational levels. the existence of a dna motif for the aspergillus nidulans ph responsive transcription factor pacc ...201222952943
conserved and essential transcription factors for cellulase gene expression in ascomycete fungi.rational engineering of filamentous fungi for improved cellulase production is hampered by our incomplete knowledge of transcriptional regulatory networks. we therefore used the model filamentous fungus neurospora crassa to search for uncharacterized transcription factors associated with cellulose deconstruction. a screen of a n. crassa transcription factor deletion collection identified two uncharacterized zinc binuclear cluster transcription factors (clr-1 and clr-2) that were required for gro ...201222532664
a mitochondrial reading frame which may code for a second form of atpase subunit 9 in aspergillus nidulans.the nucleotide sequence of a 74 codon reading frame from the aspergillus nidulans mitochondrial genome is presented. the derived amino acid sequence displays typical features of dicyclohexylcarbodiimide (dccd) binding proteins and is 84% homologous with a mitochondrial reading frame that potentially encodes an atpase subunit 9 polypeptide in neurospora crassa. however, in a. nidulans, as in n. crassa, there is strong biochemical and genetic evidence that this subunit is in fact nuclearly-encoded ...198424177948
genetic map of mitochondrial dna in podospora anserina.in order to develop an eukaryotic vector with the podospora plasmid, further characterization is required of the mitochondrial dna into which this plasmid is integrated, a physical map (restriction sites) of the podospora chondriome (size 95 kb) has been completed. as prerequisite for the establishment of a genetic (functional) map, 70% of the chondriome was cloned in e. coli vectors. using mitochondrial genes from saccharomyces cerevisiae, six structural genes were located on the podospora chon ...198224186230
high frequency transfer of species specific mitochondrial dna sequences between members of the aspergillaceae.the mitochondrial genome of aspergillus nidulans var. echinulatus is approximately 20% larger than that of the closely related species aspergillus nidulans (eidam) winter. restriction enzyme mapping and electron microscopy has revealed that the size difference is due to the presence of six inserted sequences in the former. with the exception of a small number of species specific restriction sites and the six insertions/deletions, the two mitochondrial genomes appear identical. protoplast fusion ...198124190134
identification of a novel l-rhamnose uptake transporter in the filamentous fungus aspergillus niger.the study of plant biomass utilization by fungi is a research field of great interest due to its many implications in ecology, agriculture and biotechnology. most of the efforts done to increase the understanding of the use of plant cell walls by fungi have been focused on the degradation of cellulose and hemicellulose, and transport and metabolism of their constituent monosaccharides. pectin is another important constituent of plant cell walls, but has received less attention. in relation to th ...201627984587
the introduction of the fungal d-galacturonate pathway enables the consumption of d-galacturonic acid by saccharomyces cerevisiae.pectin-rich wastes, such as citrus pulp and sugar beet pulp, are produced in considerable amounts by the juice and sugar industry and could be used as raw materials for biorefineries. one possible process in such biorefineries is the hydrolysis of these wastes and the subsequent production of ethanol. however, the ethanol-producing organism of choice, saccharomyces cerevisiae, is not able to catabolize d-galacturonic acid, which represents a considerable amount of the sugars in the hydrolysate, ...201627538689
exploring the potential of fungal arylacetonitrilases in mandelic acid synthesis.the application of arylacetonitrilases from filamentous fungi to the hydrolysis of high concentrations of (r,s)-mandelonitrile (100-500 mm) was demonstrated for the first time. escherichia coli strains expressing the corresponding genes were used as whole-cell catalysts. nitrilases from aspergillus niger, neurospora crassa, nectria haematococca, and arthroderma benhamiae (enzymes nitan, nitnc, nitnh, and nitab, respectively) exhibited different degrees of enantio- and chemoselectivity (amide for ...201525652193
fungal type iii polyketide synthases.this article covers the literature on fungal type iii polyketide synthases (pkss) published from 2005 to 2014. since the first discovery of fungal type iii pks genes in aspergillus oryzae, reported in 2005, putative genes for type iii pkss have been discovered in fungal genomes. compared with type i pkss, type iii pkss are much less abundant in fungi. however, type iii pkss could have some critical roles in fungi. this article summarizes the studies on fungal type iii pks functional analysis, in ...201425182423
a comparative study of nitrilases identified by genome mining.escherichia coli strains expressing different nitrilases transformed nitriles or kcn. six nitrilases (from aspergillus niger (2), a. oryzae, neurospora crassa, arthroderma benhamiae, and nectria haematococca) were arylacetonitrilases, two enzymes (from a. niger and penicillium chrysogenum) were cyanide hydratases and the others (from p. chrysogenum, p. marneffei, gibberella moniliformis, meyerozyma guilliermondi, rhodococcus rhodochrous, and r. ruber) preferred (hetero)aromatic nitriles as subst ...201323475593
characterization of cytotoxic compound from mangrove derived fungi irpex hydnoides vb4.to investigate the cytotoxic activity of endophytic fungi isolated from mangrove fungi.201123569763
molecular dissection of so (soft) protein in stress-induced aggregation and cell-to-cell interactive functions in filamentous fungal multicellularity.filamentous fungi grow by organizing multicellularity through hyphal compartmentalization and cell fusion. so (soft) protein, which was originally identified in neurospora crassa, plays distinct functional roles in cell-to-cell interactions, such as septal plugging and cell fusion. we previously reported that aoso, an aspergillus oryzae so homologue, forms aggregates at the septal pore in response to stress, as well as upon hyphal wounding. however, the functional regions that mediate the multic ...201627109373
improvement of omega-3 docosahexaenoic acid production by marine dinoflagellate crypthecodinium cohnii using rapeseed meal hydrolysate and waste molasses as feedstock.rapeseed meal and waste molasses are two important agro-industrial by-products which are produced in large quantities. in this study, solid state fermentation and fungal autolysis were performed to produce rapeseed meal hydrolysate (rmh) using fungal strains of aspergillus oryzae, penicillium oxalicum and neurospora crassa. the hydrolysate was used as fermentation feedstock for heterotrophic growth of microalga crypthecodinium cohnii that produce docosahexaenoic acid (dha). the addition of waste ...201525942565
a large nonconserved region of the tethering protein leashin is involved in regulating the position, movement, and function of woronin bodies in aspergillus oryzae.the woronin body is a pezizomycotina-specific organelle that is typically tethered to the septum, but upon hyphal wounding, it plugs the septal pore to prevent excessive cytoplasmic loss. leashin (lah) is a large woronin body tethering protein that contains highly conserved n- and c-terminal regions and a long (∼2,500-amino-acid) nonconserved middle region. as the involvement of the nonconserved region in woronin body function has not been investigated, here, we functionally characterized indivi ...201424813188
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