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neurospora crassa tox-1 gene encodes a ph- and temperature-tolerant mini-cellulase.cellulases that endure extreme conditions are essential in various industrial sectors. this study reports a mini-cellulase gene tox-1 from neurospora crassa. the gene tox-1 was cloned in escherichia coli after chimerization with the yebf gene and substitutions of certain isoleucine and valine with leucine residues. the yeast transformants could grow on rice straw-agar medium. the 44-amino acid peptide and its two mutant variants displayed potent cellulase activities in congo red assay and enzyma ...201627229865
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 fungus tremella mesenterica encodes the longest metallothionein currently known: gene, protein and metal binding characterization.fungal cu-thioneins, and among them, the paradigmatic neurospora crassa metallothionein (mt) (26 residues), were once considered as the shortest mts--the ubiquitous, versatile metal-binding proteins--among all organisms, and thus representatives of their primeval forms. nowadays, fungal mts of diverse lengths and sequence features are known, following the huge heterogeneity of the kingdom of fungi. at the opposite end of n. crassa mt, the recently reported cryptococcus neoformans cnmt1 and cnmt2 ...201626882011
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
circadian clocks and the regulation of virulence in fungi: getting up to speed.you cannot escape time. therefore, it seems wise to learn how to keep track of it and use it to your advantage. circadian clocks are molecular circuits that allow organisms to temporally coordinate a plethora of processes, including gene expression, with a close to 24h rhythm, optimizing cellular function in synchrony with daily environmental cycles. the molecular bases of these clocks have been extensively studied in the fungus neurospora crassa, providing a detailed molecular description. surp ...201627039027
the interaction of fungi with the environment orchestrated by rnai.the fungal kingdom has been key in the investigation of the biogenesis and function of small rnas (srnas). the discovery of phenomena such as quelling in neurospora crassa represents pioneering work in the identification of the main elements of the rna interference (rnai) machinery. recent discoveries in the regulatory mechanisms in some yeast and filamentous fungi are helping us reach a deeper understanding of the transcriptional and post-transcriptional gene-silencing mechanisms involved in ge ...201626932186
a novel zn2 cys6 transcription factor bcgaar regulates d-galacturonic acid utilization in botrytis cinerea.d-galacturonic acid (gala) is the most abundant monosaccharide component of pectin. previous transcriptome analysis in the plant pathogenic fungus botrytis cinerea identified eight gala-inducible genes involved in pectin decomposition, gala transport and utilization. co-expression of these genes indicates that a specific regulatory mechanism occurs in b. cinerea. in this study, promoter regions of these genes were analysed and eight conserved sequence motifs identified. the bclga1 promoter, cont ...201626691528
analysis of the association between codon optimality and mrna stability in schizosaccharomyces pombe.recent experiments have shown that codon optimality is a major determinant of mrna stability in saccharomyces cerevisiae and that this phenomenon may be conserved in escherichia coli and some metazoans, although work in neurospora crassa is not consistent with this model.201627825301
deoxycholic acid transformations catalyzed by selected filamentous fungi.more than 100 filamentous fungi strains, mostly ascomycetes and zygomycetes from different phyla, were screened for the ability to convert deoxycholic acid (dca) to valuable bile acid derivatives. along with 11 molds which fully degraded dca, several strains were revealed capable of producing cholic acid, ursocholic acid, 12-keto-lithocholic acid (12-keto-lca), 3-keto-dca, 15β-hydroxy-dca and 15β-hydroxy-12-oxo-lca as major products from dca. the last metabolite was found to be a new compound. t ...201626718089
expression of two novel β-glucosidases from chaetomium atrobrunneum in trichoderma reesei and characterization of the heterologous protein products.two novel gh3 family thermostable β-glucosidases from the filamentous fungus chaetomium atrobrunneum (cel3a and cel3b) were expressed in trichoderma reesei, purified by two-step ion exchange chromatography, and characterized. both enzymes were active over a wide range of ph as compared to neurospora crassa β-glucosidase gh3-3, which was also expressed in t. reesei and purified. the optimum temperature of both c. atrobrunneum enzymes was around 60 °c at ph 5, and both enzymes had better thermal a ...201627714589
suppressor of fusion, a fusarium oxysporum homolog of ndt80, is required for nutrient-dependent regulation of anastomosis.heterokaryon formation is an essential step in asexual recombination in fusarium oxysporum. filamentous fungi have an elaborate nonself recognition machinery to prevent formation and proliferation of heterokaryotic cells, called heterokaryon incompatibility (hi). in f. oxysporum the regulation of this machinery is not well understood. in neurospora crassa, vib-1, a putative transcription factor of the p53-like ndt80 family of transcription factors, has been identified as global regulator of hi. ...201627531696
selective enzymatic transformation to aldehydes in vivo by fungal carboxylate reductase from neurospora crassa.the enzymatic reduction of carboxylic acids is in its infancy with only a handful of biocatalysts available to this end. we have increased the spectrum of carboxylate-reducing enzymes (cars) with the sequence of a fungal car from neurospora crassa or74a (nccar). nccar was efficiently expressed in e. coli using an autoinduction protocol at low temperature. it was purified and characterized in vitro, revealing a broad substrate acceptance, a ph optimum at ph 5.5-6.0, a tm of 45 °c and inhibition b ...201627917101
the hsp90 co-chaperones sti1, aha1, and p23 regulate adaptive responses to antifungal azoles.heat shock protein 90 (hsp90) is essential for tumor progression in humans and drug resistance in fungi. however, the roles of its many co-chaperones in antifungal resistance are unknown. in this study, by susceptibility test of neurospora crassa mutants lacking each of 18 hsp90/calcineurin system member genes (including 8 hsp90 co-chaperone genes) to antifungal drugs and other stresses, we demonstrate that the hsp90 co-chaperones sti1 (hop1 in yeast), aha1, and p23 (sba1 in yeast) were required ...201627761133
directed evolution of xylose specific transporters to facilitate glucose-xylose co-utilization.a highly active xylose specific transporter without glucose inhibition is highly desirable in cost-effective production of biofuels from lignocellulosic biomass. however, currently available xylose specific transporters suffer from low overall activity and most are inhibited by glucose. in this study, we applied a directed evolution strategy to engineer the xylose specific transporter an25 from neurospora crassa with improved xylose transportation capacity. after four rounds of directed evolutio ...201626284760
the blue-light receptor cmwc-1 mediates fruit body development and secondary metabolism in cordyceps militaris.light is an essential factor for pigment formation and fruit body development in cordyceps militaris, a well-known edible and medicinal fungus. cmwc-1, a homolog of the blue-light receptor gene white collar-1 (wc-1) in neurospora crassa, was cloned from the c. militaris genome in our previous study. here, cmwc-1 gene inactivation results in thicker aerial hyphae, disordered fruit body development, a significant reduction in conidial formation, and carotenoid and cordycepin production. these char ...201626476643
selection of pairings reaching evenly across the data (spread): a simple algorithm to design maximally informative fully crossed mating experiments.we present a novel algorithm for the design of crossing experiments. the algorithm identifies a set of individuals (a 'crossing-set') from a larger pool of potential crossing-sets by maximizing the diversity of traits of interest, for example, maximizing the range of genetic and geographic distances between individuals included in the crossing-set. to calculate diversity, we use the mean nearest neighbor distance of crosses plotted in trait space. we implement our algorithm on a real dataset of ...201626419337
fungal genome sequencing: basic biology to biotechnology.the genome sequences provide a first glimpse into the genomic basis of the biological diversity of filamentous fungi and yeast. the genome sequence of the budding yeast, saccharomyces cerevisiae, with a small genome size, unicellular growth, and rich history of genetic and molecular analyses was a milestone of early genomics in the 1990s. the subsequent completion of fission yeast, schizosaccharomyces pombe and genetic model, neurospora crassa initiated a revolution in the genomics of the fungal ...201625721271
loss of hp1 causes depletion of h3k27me3 from facultative heterochromatin and gain of h3k27me2 at constitutive heterochromatin.methylated lysine 27 on histone h3 (h3k27me) marks repressed "facultative heterochromatin," including developmentally regulated genes in plants and animals. the mechanisms responsible for localization of h3k27me are largely unknown, perhaps in part because of the complexity of epigenetic regulatory networks. we used a relatively simple model organism bearing both facultative and constitutive heterochromatin, neurospora crassa, to explore possible interactions between elements of heterochromatin. ...201626537359
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
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
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
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
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
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
the n-acetylglucosamine catabolic gene cluster in trichoderma reesei is controlled by the ndt80-like transcription factor ron1.chitin is an important structural constituent of fungal cell walls composed of n-acetylglucosamine (glcnac) monosaccharides, but catabolism of glcnac has not been studied in filamentous fungi so far. in the yeast candida albicans, the genes encoding the three enzymes responsible for stepwise conversion of glcnac to fructose-6-phosphate are clustered. in this work, we analysed glcnac catabolism in ascomycete filamentous fungi and found that the respective genes are also clustered in these fungi. ...201626481444
xylitol production by saccharomyces cerevisiae overexpressing different xylose reductases using non-detoxified hemicellulosic hydrolysate of corncob.xylitol production was compared in fed batch fermentation by saccharomyces cerevisiae strains overexpressing xylose reductase (xr) genes from candida tropicalis, pichia stipitis, neurospora crassa, and an endogenous gene gre3. the gene encoding a xylose specific transporter (sut1) from p. stipitis was cloned to improve xylose transport and fed batch fermentation was used with glucose as a cosubstrate to regenerate nadph. xylitol yield was near theoretical for all the strains in fed batch ferment ...201628330197
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
[proteins from neurospora crassa. 2. immunoelectrophoretically detectable proteins and their relationship to growth time]. 20164973223
isolation, mapping, and characterization of trehalaseless mutants of neurospora crassa.mutant strains of neurospora crassa that lack trehalase and are unable to grow on trehalose were isolated, and the gene (tre) was positioned on the right arm of linkage group i. maltase and beta-galactosidase activities are almost identical in tre(-) strains, whereas that of invertase was reduced by more than half and those of acid phosphatase and amylase were somewhat increased. heterocaryons between standard and trehalaseless strains yield less than one-tenth the activity of the former. in add ...20165001211
repositioning of drugs using open-access data portal dtome: a test case with probenecid (review).the one gene-one enzyme hypothesis, first introduced by beadle and tatum in the 1940s and based on their genetic analysis and observation of phenotype changes in neurospora crassa challenged by various experimental conditions, has witnessed significant advances in recent decades. much of our understanding of the association between genes and their phenotype expression has benefited from the completion of the human genome project, and has shown continual transformation guided by the effort direct ...201626572802
fine-scale mapping in neurospora crassa by using genome-wide knockout strains.fine-scale genetic mapping is often hindered by the lack of adequate markers surrounding the locus of interest. in the filamentous ascomycete neurospora crassa, the genome has been sequenced and an effort has been made to generate genome-wide deletion strains for the entire gene set. accordingly, the hygromycin-resistant marker in each deletion strain can be used as a mapping locus in a classical three-point cross, along with the mapping target and a standard marker. we have demonstrated the fea ...201622067301
the growth of the isolated hyphal apices of neurospora crassa under carbon starvation. 201618841815
potassium distribution in the neurospora crassa hypha. 201611029033
kinesin and dynein mutants provide novel insights into the roles of vesicle traffic during cell morphogenesis in neurospora.kinesin and cytoplasmic dynein are force-generating molecules that move in opposite directions along microtubules. they have been implicated in the directed transport of a wide variety of cellular organelles, but it is unclear whether they have overlapping or largely independent functions.201610469561
melanogenesis, tyrosinase expression, and reproductive differentiation in black and white truffles (ascomycotina).white and black truffles of the genus tuber are ascomycotina as well as neurospora crassa, which expresses tyrosinase dependently on the reproductive cycle. tyrosinase expression dependent on reproductive differentiation has been also described in black truffles. we present novel and comparative work on melanogenic activities in black and white truffles that both express true tyrosinases. l-tyrosine 3-monooxygenase and l-dopa oxidase activities colocalize as histochemically detected and are simi ...20169170162
mutation at the am locus of neurospora crassa.forty-eight new mutations at the am locus of neurospora crassa have been characterized. nineteen mutations were induced by uv; of these, eight were missense, two were frameshifts, two were nonsense, three were deletions and four were unidentified. twenty-nine mutations were induced with nitrous acid; of these, twenty-one were missense, three were frameshifts, one was nonsense, two were deletions and one was genetically unstable.20166177580
[isolation and certain features of a polyphosphate phosphohydrolase deficient mutant of the fungus neurospora crassa].a polyphosphatase deficient mutant of neurospora crassa has been isolated. the criterion for selecting the mutant was the capacity of the fungus to assimilate polyphosphates as the source of exogenous phosphorus. the mutant like the parent strain ad-6, was an adenine auxotroph but differed from the parent strain by a lower growth rate though, at the stationary stage, its biomass reached the same level as in the strain ad-6. the character of changes in the activity of polyphosphatase in the cours ...20166248740
conversion of d-lysine via l-pepecolic acid in neurospora crassa.neurospora crassa converts d-lysine to l-pipecolic acid and the latter to l-lysine by a pathway outlined herein and different from mechanisms hitherto known to be involved in interconversions of enantiomers of amino acids.2016125972
[isolation and properties of polyphosphatase of neurospora crassa].polyphosphatase (polyphosphate-phosphohydrolase) has been isolated from mycelium of neurospora crassa and purified to homogenous state. the enzyme is shown to be strictly specific to high molecular weight inorganic polyphosphates. km for phosphate in polymeric form is 6.8-10(-4) m. the molecular weight of this enzyme is 50 000 +/- 3000. to display its activity polyphosphatase requires the presence of bivalent cations of some metals, mg2+ ions being the best activator with co2+, mn2+ and fe2+ ion ...20162861
[polyphosphate depolymerase activity in cells of neurospora crassa (28610 as)]. 20164344204
[location of polyphosphatases, hydrolyzing polyphosphates to orthophosphates, in the subcellular structures of neurospora crassa]. 20164336095
[detection and some properties of neurospora crassa polyphosphatases hydrolyzing inorganic polyphosphates and orthophosphates]. 20164333905
ageing of neurospora crassa. iv. induction of senescence in wild type by dietary amino acid analogs and reversal by antioxidants and membrane stabilizers.the extensional growth rate of wild-type 74a8 n. crassa in the presence of various concentrations of 19 amino acid analogs was measured. seven analogs were not inhibitory at concentrations in the range of one to 10mm. of the remaining 12 analogs, nine inhibited growth in a novel way. the kinetics of growth in the presence of these analogs at 30 degrees were characterized by seven sequential phases: (1)lag; (2) acceleration of growth rate; (3) steady-state growth rate; (4) exponential rate of dec ...20157715
purification of the 2-oxoglutarate dehydrogenase and pyruvate dehydrogenase complexes of neurospora crassa mitochondria.a simple purification procedure for the 2-oxoglutarate dehydrogenase and the pyruvate dehydrogenase complexes of neurospora crassa mitochondria is described. after fractionated precipitations with polyethylene glycol, elimination of thiol proteins, and gel-filtration chromatography, the resulting preparations contained both activities. covalent chromatography on thiol-activated sepharose cl-4b allowed the specific binding of the 2-oxoglutarate dehydrogenase complex activity in the presence of 2- ...20152947635
dna-dna and dna-protein crosslinking and repair in neurospora crassa following exposure to nitrogen mustard.a new method was developed for the analysis of dna-protein crosslinks in neurospora crassa. the formations of dna-protein and dna-dna crosslinks were assayed following exposure of spheroplasts to hn2. both types of crosslink were detected and were found to be repaired during recovery. moreover a mutant sensitive to hn2 was defective in the removal of both types of crosslink.20156239978
cold-induced increase of glycerol kinase in neurospora crassa. 20154270897
induction of beta-glucosidases in neurospora crassa.the induction of beta-glucosidases (ec 3.2.1.21) was studied in neurospora crassa. cellobiase was induced by cellobiose, but other inducers had little effect on this enzyme. cellobiase activity was very low in all stages of the vegetative life cycle in the absence of di-beta-glucoside inducer. aryl-beta-glucosidase was semiconstitutive at late stages of culture growth prior to conidiation. at early stages, aryl-beta-glucosidase was induced by cellobiose, laminaribiose, and gentiobiose, and weakl ...20154270443
studies on the glutamate dehydrogenase from neurospora crassa. 20154403951
isolation and characterization of an unsaponifiable lipid from neurospora crassa. 20154364989
cyclic amp-induced tyrosinase synthesis in neurospora crassa. 20154376002
chemically induced reversion and suppression of methionine-i alleles of neurospora crassa. 20154357302
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
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
the subunit structure of the nicotinamide-adenine dinucleotide phosphate-dependent glutamate dehydrogenase of neurospora crassa. 20154403952
behavior of cycloaldolase from neurospora crassa towards substrate analogs and aldolase inhibitors. 20154346519
evidence for distinct kynureninase and hydroxykynureninase activities in neurospora crassa.previous studies have indicated that a single enzyme, "kynureninase," catalyzes the reactions of l-kynurenine to anthranilate and l-3-hydroxykynurenine to 3-hydroxyanthranilate in neurospora crassa and in other organisms. the present report describes separate enzymes which catalyze these reactions in n. crassa. the first, a kynureninase, preferentially catalyzes kynurenine to anthranilate and is induced over 400-fold by tryptophan or a catabolite of tryptophan. the second, a hydroxykynureninase, ...20154331500
regulation of nitrate reductase in neurospora crassa: stability in vivo.nicotinamide adenine dinucleotide phosphate, reduced form (nadph)-nitrate reductase and its related enzyme activities, nadph-cytochrome c reductase and reduced benzyl viologen-nitrate reductase, are all induced following the transfer of ammonia-grown wild-type neurospora mycelia to nitrate medium. after nitrate reductase is induced to the maximal level, the addition of an ammonium salt to, or the removal of nitrate from, the cultures results in a rapid inactivation of nitrate reductase and its t ...20154401813
comparison of cysteine and tryptophan content of insoluble proteins derived from wild-type and mi-1 strains of neurospora crassa.the possibility of an amino acid substitution (cysteine for tryptophan) in a membrane protein of the [mi-1] strain of neurospora crassa has been investigated in detail by using a double radioactive labeling procedure. auxotrophic strains of neurospora having wild-type [+] or [mi-1] cytoplasm have been grown under conditions which result in the specific labeling of protein tryptophan with (3)h and protein cysteine with (35)s. although the least soluble 1 to 20% of the [mi-1] mitochondrial membran ...20154334762
formation of assimilatory nitrate reductase by in vitro inter-cistronic complementation in neurospora crassa.in vitro complementation of the soluble assimilatory nicotinamide adenine dinucleotide phosphate, reduced form (nadph)-nitrate reductase was attained by mixing cell-free preparations of certain neurospora nitrate reductase mutants: induced nit-1 (uniquely possessing inducible nadph-cytochrome c reductase) with (a) uninduced or induced nit-2 or nit-3, or (b) uninduced wild type. the complementing activity of induced nit-1 is soluble while that of nit-2, nit-3, and wild type is particulate but not ...20154391854
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
anti-infective assessment of senecio smithioides (asteraceae) and isolation of 9-oxoeuryopsin, a furanoeremophilane-type sesquiterpene with antiplasmodial activity.the search for anti-infective activity in the antipyretic plant senecio smithioides was conducted. petroleum ether (pe), dichloromethane (ch2cl2), ethyl acetate (etoac) and hydroethanolic (96% etoh) extracts, and compounds 9-oxoeuryopsin (1), epoxydecompostin (2) and senecionine (3) were obtained from the aerial parts. all extracts and 1 were tested against chloroquine-resistant strain of plasmodium falciparum (ref. chloroquine), trypanosoma cruzi (ref. nifurtimox), leishmania braziliensis, leis ...201526606687
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
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
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
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
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
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
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
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
[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
compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics.interrogating polarized growth is technologically challenging due to extensive cellular branching and uncontrollable environmental conditions in conventional assays. here we present a robust and high-performance microfluidic system that enables observations of polarized growth with enhanced temporal and spatial control over prolonged periods. the system has built-in tunability and versatility to accommodate a variety of scientific applications requiring precisely controlled environments. using t ...201526530004
transcription profiling of the neurospora crassa response to a group of synthetic (thio)xanthones and a natural acetophenone.xanthones are a class of heterocyclic compounds characterized by a dibenzo-γ-pyrone nucleus. analysis of their mode of action in cells, namely uncovering alterations in gene expression, is important because these compounds have potential therapeutic applications. thus, we studied the transcriptional response of the filamentous fungus neurospora crassa to a group of synthetic (thio)xanthone derivatives with antitumor activity using high throughput rna sequencing. the induction of abc transporters ...201526484172
the chestnut blight fungus for studies on virus/host and virus/virus interactions: from a natural to a model host.the chestnut blight fungus, cryphonectria parasitica, is an important plant pathogenic ascomycete. the fungus hosts a wide range of viruses and now has been established as a model filamentous fungus for studying virus/host and virus/virus interactions. this is based on the development of methods for artificial virus introduction and elimination, host genome manipulability, available host genome sequence with annotations, host mutant strains, and molecular tools. molecular tools include sub-cellu ...201525454384
prevalent and distinct spliceosomal 3'-end processing mechanisms for fungal telomerase rna.telomerase rna (ter) is an essential component of the telomerase ribonucleoprotein complex. the mechanism for ter 3'-end processing is highly divergent among different organisms. here we report a unique spliceosome-mediated ter 3'-end cleavage mechanism in neurospora crassa that is distinct from that found specifically in the fission yeast schizosaccharomyces pombe. while the s. pombe ter intron contains the canonical 5'-splice site guaugu, the n. crassa ter intron contains a non-canonical 5'-sp ...201525598218
identification and characterization of lfd-2, a predicted fringe protein required for membrane integrity during cell fusion in neurospora crassa.the molecular mechanisms of membrane merger during somatic cell fusion in eukaryotic species are poorly understood. in the filamentous fungus neurospora crassa, somatic cell fusion occurs between genetically identical germinated asexual spores (germlings) and between hyphae to form the interconnected network characteristic of a filamentous fungal colony. in n. crassa, two proteins have been identified to function at the step of membrane fusion during somatic cell fusion: prm1 and lfd-1. the abse ...201525595444
hypotonic stress-induced calcium signaling in saccharomyces cerevisiae involves trp-like transporters on the endoplasmic reticulum membrane.saccharomyces cerevisiae cells respond to hypotonic stress (hts) by a cytosolic calcium rise, either generated by an influx of calcium from extracellular medium, when calcium is available, or by a release from intracellular stores in scarcity of extracellular calcium. calcium release from intracellular compartments is peculiarly inhibited by external calcium in a calcineurin-independent and cch1-, but not mid1-, driven manner. hts-induced calcium release is also negatively regulated by the er pr ...201525573187
mechanism of the neurospora circadian clock, a frequency-centric view.circadian clocks are self-sustaining timekeepers found in almost all organisms on earth. the filamentous fungus neurospora crassa is a preeminent model for eukaryotic circadian clocks. investigations of the neurospora circadian clock system have led to elucidation of circadian clock regulatory mechanisms that are common to all eukaryotes. in this work, we will focus on the neurospora circadian oscillator mechanism with an emphasis on the regulation of the core clock component frequency.201525302868
the rab gtpase ypt-1 associates with golgi cisternae and spitzenkörper microvesicles in neurospora crassa.vesicle traffic involves budding, transport, tethering and fusion of vesicles with acceptor membranes. gtp-bound small rab gtpases interact with the membrane of vesicles, promoting their association with other factors before their subsequent fusion. filamentous fungi contain at their hyphal apex the spitzenkörper (spk), a multivesicular structure to which vesicles concentrate before being redirected to specific cell sites. the regulatory mechanisms ensuring the directionality of the vesicles tha ...201525425138
development and physiological characterization of cellobiose-consuming yarrowia lipolytica.yarrowia lipolytica is a promising production host for a wide range of molecules, but limited sugar consumption abilities prevent utilization of an abundant source of renewable feedstocks. in this study we created a y. lipolytica strain capable of utilizing cellobiose as a sole carbon source by using endogenous promoters to express the cellodextrin transporter cdt-1 and intracellular β-glucosidase gh1-1 from neurospora crassa. the engineered strain was also capable of simultaneous co-consumption ...201525421388
the putative cellodextrin transporter-like protein clp1 is involved in cellulase induction in neurospora crassa.neurospora crassa recently has become a novel system to investigate cellulase induction. here, we discovered a novel membrane protein, cellodextrin transporter-like protein 1 (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 c ...201525398875
functional characterization and low-resolution structure of an endoglucanase cel45a from the filamentous fungus neurospora crassa or74a: thermostable enzyme with high activity toward lichenan and β-glucan.biomass is the most abundant and short-term renewable natural resource on earth whose recalcitrance toward enzymatic degradation represents significant challenge for a number of biotechnological applications. the not so abundant but critically necessary class of gh45 endoglucanases constitutes an essential component of tailored industrial enzyme cocktails because they randomly and internally cleave cellulose molecules. moreover, gh45 glucanases are core constituents of major-brand detergent form ...201525711741
heterologous expression of mitochondria-targeted microbial nitrilase enzymes increases cyanide tolerance in arabidopsis.anthropogenic activities have resulted in cyanide (cn) contamination of both soil and water in many areas of the globe. while plants possess a detoxification pathway that serves to degrade endogenously generated cn, this system is readily overwhelmed, limiting the use of plants in bioremediation. genetic engineering of additional cn degradation pathways in plants is one potential strategy to increase their tolerance to cn. here we show that heterologous expression of microbial nitrilase enzymes ...201525711239
cell fusion in neurospora crassa.in recent years, the filamentous fungus neurospora crassa has advanced as a model organism for studying eukaryotic cell-cell communication and fusion. cell merger in this fungus employs an unusual mode of communication, in which the fusion partners appear to switch between signal sending and receiving. many molecular factors mediating this intriguing mechanism and the subsequent membrane merger have been identified. it has become apparent that conserved factors, such as map kinases, nadph oxidas ...201526340439
paramutation phenomena in plants.paramutation is a particular epigenetic phenomenon discovered in zea mays by alexander brink in the 1950s, and then also found in other plants and animals. brink coined the term paramutation (from the greek syllable "para" meaning beside, near, beyond, aside) in 1958, with the aim to differentiate paramutation from mutation. the peculiarity of paramutation with respect to other gene silencing phenomena consists in the ability of the silenced allele (named paramutagenic) to silence the other alle ...201526335267
don't go with the cytoplasmic flow.cytoplasmic streaming occurs in a variety of organisms, but its biological function is often unclear. in this issue of developmental cell, pieuchot et al. (2015) report that streaming creates subcellular compartments in the filamentous fungus neurospora crassa. nuclei immobilized within these compartments differentiate and may drive a compartment-specific genetic program.201526305589
involvement of the adaptor protein 3 complex in lignocellulase secretion in neurospora crassa revealed by comparative genomic screening.lignocellulase hypersecretion has been achieved in industrial fungal workhorses such as trichoderma reesei, but the underlying mechanism associated with this process is not well understood. although previous comparative genomic studies have revealed that the mutagenic t. reesei strain rut-c30 harbors hundreds of mutations compared with its parental strain qm6a, how these mutations actually contribute to the hypersecretion phenotype remains to be elucidated.201526300971
role of bgt-1 and bgt-2, two predicted gpi-anchored glycoside hydrolases/glycosyltransferases, in cell wall remodeling in neurospora crassa.neurospora crassa bgt-1 (ncu06381) and bgt-2 (ncu09175) are two putative glycoside hydrolases (ghs) with additional predicted glycosyltransferase activity and binding sites for a glycosyl phosphatidyl inositol (gpi) anchor that would facilitate their attachment to the plasma membrane (pm). to discern their role in key morphogenetic events during vegetative development of n. crassa, bgt-1 and bgt-2 were labeled with the green fluorescent protein (gfp). the gfp was inserted immediately after the s ...201526541633
global analysis of predicted g protein-coupled receptor genes in the filamentous fungus, neurospora crassa.g protein-coupled receptors (gpcrs) regulate facets of growth, development, and environmental sensing in eukaryotes, including filamentous fungi. the largest predicted gpcr class in these organisms is the pth11-related, with members similar to a protein required for disease in the plant pathogen magnaporthe oryzae. however, the pth11-related class has not been functionally studied in any filamentous fungal species. here, we analyze phenotypes in available mutants for 36 gpcr genes, including 20 ...201526464358
epigenetic and posttranslational modifications in light signal transduction and the circadian clock in neurospora crassa.blue light, a key abiotic signal, regulates a wide variety of physiological processes in many organisms. one of these phenomena is the circadian rhythm presents in organisms sensitive to the phase-setting effects of blue light and under control of the daily alternation of light and dark. circadian clocks consist of autoregulatory alternating negative and positive feedback loops intimately connected with the cellular metabolism and biochemical processes. neurospora crassa provides an excellent mo ...201526198228
metabolic impacts of using nitrogen and copper-regulated promoters to regulate gene expression in neurospora crassa.the filamentous fungus neurospora crassa is a long-studied eukaryotic microbial system amenable to heterologous expression of native and foreign proteins. however, relatively few highly tunable promoters have been developed for this species. in this study, we compare the tcu-1 and nit-6 promoters for controlled expression of a gfp reporter gene in n. crassa. although the copper-regulated tcu-1 has been previously characterized, this is the first investigation exploring nitrogen-controlled nit-6 ...201526194204
structural and functional characterization of a lytic polysaccharide monooxygenase with broad substrate specificity.the recently discovered lytic polysaccharide monooxygenases (lpmos) carry out oxidative cleavage of polysaccharides and are of major importance for efficient processing of biomass. nclpmo9c from neurospora crassa acts both on cellulose and on non-cellulose β-glucans, including cellodextrins and xyloglucan. the crystal structure of the catalytic domain of nclpmo9c revealed an extended, highly polar substrate-binding surface well suited to interact with a variety of sugar substrates. the ability o ...201526178376
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