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the neurospora crassa exocyst complex tethers spitzenkörper vesicles to the apical plasma membrane during polarized growth.fungal hyphae are among the most highly polarized cells. hyphal polarized growth is supported by tip-directed transport of secretory vesicles, which accumulate temporarily in a stratified manner in an apical vesicle cluster, the spitzenkörper. the exocyst complex is required for tethering of secretory vesicles to the apical plasma membrane. we determined that the presence of an octameric exocyst complex is required for the formation of a functional spitzenkörper and maintenance of regular hyphal ...201424523289
an in vitro peptide complementation assay for cyt-18-dependent group i intron splicing reveals a new role for the n-terminus.the mitochondrial tyrosyl trna synthetase from neurospora crassa (cyt-18 protein) is a bifunctional group i intron splicing cofactor. cyt-18 is capable of splicing multiple group i introns from a wide variety of sources by stabilizing the catalytically active intron structures. cyt-18 and mt tyrrss from related fungal species have evolved to assist in group i intron splicing in part by the accumulation of three n-terminal domain insertions. biochemical and structural analysis indicate that the n ...201424520960
identification and characterization of a galacturonic acid transporter from neurospora crassa and its application for saccharomyces cerevisiae fermentation processes.pectin-rich agricultural wastes potentially represent favorable feedstocks for the sustainable production of alternative energy and bio-products. their efficient utilization requires the conversion of all major constituent sugars. the current inability of the popular fermentation host saccharomyces cerevisiae to metabolize the major pectic monosaccharide d-galacturonic acid (d-gala) significantly hampers these efforts. while it has been reasoned that the optimization of cellular d-gala uptake wi ...201424502254
the proteome and phosphoproteome of neurospora crassa in response to cellulose, sucrose and carbon starvation.improving cellulolytic enzyme production by plant biomass degrading fungi holds great potential in reducing costs associated with production of next-generation biofuels generated from lignocellulose. how fungi sense cellulosic materials and respond by secreting enzymes has mainly been examined by assessing function of transcriptional regulators and via transcriptional profiling. here, we obtained global proteomic and phosphoproteomic profiles of the plant biomass degrading filamentous fungus neu ...201424881580
an asymmetric pan3 dimer recruits a single pan2 exonuclease to mediate mrna deadenylation and decay.the pan2-pan3 complex functions in general and microrna-mediated mrna deadenylation. however, mechanistic insight into pan2 and its complex with the asymmetric pan3 dimer is lacking. here, we describe crystal structures that show that neurospora crassa pan2 comprises two independent structural units: a c-terminal catalytic unit and an n-terminal assembly unit that engages in a bipartite interaction with pan3 dimers. the catalytic unit contains the exonuclease domain in an intimate complex with a ...201424880343
heterochromatin controls γh2a localization in neurospora crassa.in response to genotoxic stress, atr and atm kinases phosphorylate h2a in fungi and h2ax in animals on a c-terminal serine. the resulting modified histone, called γh2a, recruits chromatin-binding proteins that stabilize stalled replication forks or promote dna double-strand-break repair. to identify genomic loci that might be prone to replication fork stalling or dna breakage in neurospora crassa, we performed chromatin immunoprecipitation (chip) of γh2a followed by next-generation sequencing (c ...201424879124
secret lifestyles of neurospora crassa.neurospora crassa has a long history as an excellent model for genetic, cellular, and biochemical research. although this fungus is known as a saprotroph, it normally appears on burned vegetations or trees after forest fires. however, due to a lack of experimental evidence, the nature of its association with living plants remains enigmatic. here we report that scots pine (pinus sylvestris) is a host plant for n. crassa. the endophytic lifestyle of n. crassa was found in its interaction with scot ...201424875794
experimental evolution of bet hedging under manipulated environmental uncertainty in neurospora crassa.all organisms are faced with environmental uncertainty. bet-hedging theory expects unpredictable selection to result in the evolution of traits that maximize the geometric-mean fitness even though such traits appear to be detrimental over the shorter term. despite the centrality of fitness measures to evolutionary analysis, no direct test of the geometric-mean fitness principle exists. here, we directly distinguish between predictions of competing fitness maximization principles by testing cohen ...201424870047
dynamics of transcriptome evolution in the model eukaryote neurospora.mounting evidence indicates that changes in the transcriptome contribute significantly to the phenotypic differentiation of closely related species. nonetheless, further genome-wide studies, spanning a broad range of organisms, are needed to decipher the factors driving transcriptome evolution. the model neurospora (ascomycota) comprises a simple system for empirically studying the evolutionary dynamics of the transcriptome. here, we studied the evolution of gene expression in neurospora crassa ...201424848562
neurospora crassa 1,3-α-glucan synthase, ags-1, is required for cell wall biosynthesis during macroconidia development.the neurospora crassa genome encodes two 1,3-α-glucan synthases. one of these 1,3-α-glucan synthase genes, ags-1, was shown to be required for the synthesis of 1,3-α-glucan in the aerial hyphae and macroconidia cell walls. 1,3-α-glucan was found in the conidia cell wall, but was absent from the vegetative hyphae cell wall. deletion of ags-1 affected conidial development. δags-1 produced only 5 % as many conidia as the wt and most of the conidia produced by δags-1 were not viable. the ags-1 upstr ...201424847001
[improving cellulases production with neurospora crassa by morphology mutants screening].filamentous fungi are widely used for large-scale production of cellulases. morphological characteristics of mycelia under submerged condition are closely correlated with cellulases productivity. in order to find out the critical genes involved in the mycelial morphology development and cellulases production in liquid fermentation, 95 neurospora crassa morphological mutants (named as szy1-95) were screened for cellulases production. compared with the wild type, cellulases production in four muta ...201424818479
directed evolution of a cellodextrin transporter for improved biofuel production under anaerobic conditions in saccharomyces cerevisiae.introduction of a cellobiose utilization pathway consisting of a cellodextrin transporter and a β-glucosidase into saccharomyces cerevisiae enables co-fermentation of cellobiose and xylose. cellodextrin transporter 1 (cdt1) from neurospora crassa has been established as an effective transporter for the engineered cellobiose utilization pathways. however, cellodextrin transporter 2 (cdt2) from the same species is a facilitator and has the potential to be more efficient than cdt1 under anaerobic c ...201424519319
septum development in neurospora crassa: the septal actomyosin tangle.septum formation in neurospora crassa was studied by fluorescent tagging of actin, myosin, tropomyosin, formin, fimbrin, bud-4, and chs-1. in chronological order, we recognized three septum development stages: 1) septal actomyosin tangle (sat) assembly, 2) contractile actomyosin ring (car) formation, 3) car constriction together with plasma membrane ingrowth and cell wall construction. septation began with the assembly of a conspicuous tangle of cortical actin cables (sat) in the septation site ...201424800890
analysis of the antimicrobial effects of nonthermal plasma on fungal spores in ionic solutions.the antimicrobial efficiency of reactive species-based control strategies is significantly affected by the dynamics of reactive species in the biological environment. atmospheric-pressure nonthermal plasma is an ionized gas in which various reactive species are produced. the various levels of antimicrobial activity may result from the dynamic interaction of the plasma-generated reactive species with the environment. however, the nature of the interaction between plasma and environments is poorly ...201424794411
cdc-42 and rac-1 regulate opposite chemotropisms in neurospora crassa.cell polarization and fusion are crucial developmental processes that occur in response to intracellular and extracellular signals. asexual spores (conidia) of the mold neurospora crassa differentiate two types of polarized cell protrusions, germ tubes and conidial anastomosis tubes (cats), which exhibit negative and positive chemotropism, respectively. we provide the first evidence that shared and separate functions of the rho-type gtpases cdc-42 and rac-1 regulate these opposite chemotropisms. ...201424790223
specific domains of plant defensins differentially disrupt colony initiation, cell fusion and calcium homeostasis in neurospora crassa.msdef1 and mtdef4 from medicago spp. are small cysteine-rich defensins with potent antifungal activity against a broad range of filamentous fungi. each defensin has a hallmark γ-core motif (gxcx(3-9) c), which contains major determinants of its antifungal activity. in this study, the antifungal activities of msdef1, mtdef4, and peptides derived from their γ-core motifs, were characterized during colony initiation in the fungal model, neurospora crassa. these defensins and their cognate peptides ...201424773060
proper actin ring formation and septum constriction requires coordinated regulation of sin and mor pathways through the germinal centre kinase mst-1.nuclear dbf2p-related (ndr) kinases constitute a functionally conserved protein family of eukaryotic regulators that control cell division and polarity. in fungi, they function as effector kinases of the morphogenesis (mor) and septation initiation (sin) networks and are activated by pathway-specific germinal centre (gc) kinases. we characterized a third gc kinase, mst-1, that connects both kinase cascades. genetic and biochemical interactions with sin components and life cell imaging identify m ...201424762679
alteration of light-dependent gene regulation by the absence of the rco-1/rcm-1 repressor complex in the fungus neurospora crassa.the activation of transcription by light in the fungus neurospora crassa requires the white collar complex (wcc), a photoreceptor and transcription factor complex. after light reception two wccs interact and bind the promoters of light-regulated genes to activate transcription. this process is regulated by vvd, a small photoreceptor that disrupts the interaction between wccs and leads to a reduction in transcription after long exposures to light. the n. crassa rco-1/rcm-1 repressor complex is th ...201424747913
discovery of lpmo activity on hemicelluloses shows the importance of oxidative processes in plant cell wall degradation.the recently discovered lytic polysaccharide monooxygenases (lpmos) are known to carry out oxidative cleavage of glycoside bonds in chitin and cellulose, thus boosting the activity of well-known hydrolytic depolymerizing enzymes. because biomass-degrading microorganisms tend to produce a plethora of lpmos, and considering the complexity and copolymeric nature of the plant cell wall, it has been speculated that some lpmos may act on other substrates, in particular the hemicelluloses that tether t ...201424733907
the highly expressed methionine synthase gene of neurospora crassa is positively regulated by its proximal heterochromatic region.in neurospora crassa, the methionine synthase gene met-8 plays a key role in methionine synthesis. in this study, we found that met-8 protein levels were compromised in several mutants defective in proper heterochromatin formation. bioinformatics analysis revealed a 50-kb at-rich region adjacent to the met-8 promoter. chip assays confirmed that trimethylated h3k9 was enriched in this region, indicating that heterochromatin may form upstream of met-8. in an h3k9r mutant strain, the output of met- ...201424711369
the rna helicase frh is an atp-dependent regulator of ck1a in the circadian clock of neurospora crassa.the neurospora clock protein frq forms a complex with casein kinase 1a (ck1a) and frh, a dead box-containing rna helicase with a clock-independent essential function in rna metabolism. in the course of a circadian period, frq is progressively hyperphosphorylated and eventually degraded. timed hyperphosphorylation of frq is crucial for timekeeping of the clock. here we show that the atpase activity of frh attenuates the kinetics of ck1a-mediated hyperphosphorylation of frq. hyperphosphorylation o ...201424710172
engineering the filamentous fungus neurospora crassa for lipid production from lignocellulosic biomass.microbially produced triacylglycerol (tag) is a potential feedstock for the production of biodiesel, but its commercialization will require high yields from low-cost renewable feedstocks such as lignocellulose. the present study employs a multi-gene approach to increasing tag biosynthesis in the filamentous fungus neurospora crassa. we demonstrate the redirection of carbon flux from glycogen biosynthesis towards fatty acid biosynthesis in a glycogen synthase deletion strain (δgsy-1). furthermore ...201424700367
crystallization and preliminary x-ray crystallographic analysis of importin-α from neurospora crassa.importin-α recognizes cargo proteins that contain classical nuclear localization sequences (nls) and, in complex with importin-β, is able to translocate nuclear proteins through the nuclear pore complex. the filamentous fungus neurospora crassa is a well studied organism that has been widely used as a model organism for fundamental aspects of eukaryotic biology, and is important for understanding the specific mechanisms of protein transport to the cell nucleus. in this work, the crystallization ...201424699749
direct recognition of homology between double helices of dna in neurospora crassa.chromosomal regions of identical or nearly identical dna sequence can preferentially associate with one another in the apparent absence of dna breakage. molecular mechanism(s) underlying such homology-dependent pairing phenomena remain(s) unknown. using neurospora crassa repeat-induced point mutation (rip) as a model system, we show that a pair of dna segments can be recognized as homologous, if they share triplets of base pairs arrayed with the matching periodicity of 11 or 12 base pairs. this ...201424699390
discovering functions of unannotated genes from a transcriptome survey of wild fungal isolates.most fungal genomes are poorly annotated, and many fungal traits of industrial and biomedical relevance are not well suited to classical genetic screens. assigning genes to phenotypes on a genomic scale thus remains an urgent need in the field. we developed an approach to infer gene function from expression profiles of wild fungal isolates, and we applied our strategy to the filamentous fungus neurospora crassa. using transcriptome measurements in 70 strains from two well-defined clades of this ...201424692637
fungal secretomes enhance sugar beet pulp hydrolysis.the recalcitrance of lignocellulose makes enzymatic hydrolysis of plant biomass for the production of second generation biofuels a major challenge. this work investigates an efficient and economic approach for the enzymatic hydrolysis of sugar beet pulp (sbp), which is a difficult to degrade, hemicellulose-rich by-product of the table sugar industry. three fungal strains were grown on different substrates and the production of various extracellular hydrolytic and oxidative enzymes involved in pe ...201424677771
identification and characterization of lfd1, a novel protein involved in membrane merger during cell fusion in neurospora crassa.despite its essential role in development, molecular mechanisms of membrane merger during cell-cell fusion in most eukaryotic organisms remain elusive. in the filamentous fungus neurospora crassa, cell fusion occurs during asexual spore germination, where genetically identical germlings show chemotropic interactions and cell-cell fusion. fusion of germlings and hyphae is required for the formation of the interconnected mycelial network characteristic of filamentous fungi. previously, a multipass ...201424673848
cellulose surface degradation by a lytic polysaccharide monooxygenase and its effect on cellulase hydrolytic efficiency.lytic polysaccharide monooxygenase (lpmo) represents a unique principle of oxidative degradation of recalcitrant insoluble polysaccharides. used in combination with hydrolytic enzymes, lpmo appears to constitute a significant factor of the efficiency of enzymatic biomass depolymerization. lpmo activity on different cellulose substrates has been shown from the slow release of oxidized oligosaccharides into solution, but an immediate and direct demonstration of the enzyme action on the cellulose s ...201425361767
the transcription factor ste12 mediates the regulatory role of the tmk1 map kinase in mycoparasitism and vegetative hyphal fusion in the filamentous fungus trichoderma atroviride.mycoparasitic species of the fungal genus trichoderma are potent antagonists able to combat plant pathogenic fungi by direct parasitism. an essential step in this mycoparasitic fungus-fungus interaction is the detection of the fungal host followed by activation of molecular weapons in the mycoparasite by host-derived signals. the trichoderma atroviride map kinase tmk1, a homolog of yeast fus3/kss1, plays an essential role in regulating the mycoparasitic host attack, aerial hyphae formation and c ...201425356841
gene expression differences among three neurospora species reveal genes required for sexual reproduction in neurospora crassa.many fungi form complex three-dimensional fruiting bodies, within which the meiotic machinery for sexual spore production has been considered to be largely conserved over evolutionary time. indeed, much of what we know about meiosis in plant and animal taxa has been deeply informed by studies of meiosis in saccharomyces and neurospora. nevertheless, the genetic basis of fruiting body development and its regulation in relation to meiosis in fungi is barely known, even within the best studied mult ...201425329823
isolation, characterization and application of a cellulose-degrading strain neurospora crassa s1 from oil palm empty fruit bunch.oil palm empty fruit bunch (efb) is a lignocellulosic waste produced in palm oil industry. efb mainly consists of cellulose, hemicellulose (mainly xylan) and lignin and has a great potential to be reused. converting efb to fermentable sugars and value-added chemicals is a much better choice than treating efb as waste.201425384340
ham-5 functions as a map kinase scaffold during cell fusion in neurospora crassa.cell fusion in genetically identical neurospora crassa germlings and in hyphae is a highly regulated process involving the activation of a conserved map kinase cascade that includes nrc-1, mek-2 and mak-2. during chemotrophic growth in germlings, the map kinase cascade members localize to conidial anastomosis tube (cat) tips every ∼8 minutes, perfectly out of phase with another protein that is recruited to the tip: soft, a recently identified scaffold for the mak-1 map kinase pathway in sordaria ...201425412208
fungal communication requires the mak-2 pathway elements ste-20 and ras-2, the nrc-1 adapter ste-50 and the map kinase scaffold ham-5.intercellular communication is critical for the survival of unicellular organisms as well as for the development and function of multicellular tissues. cell-to-cell signaling is also required to develop the interconnected mycelial network characteristic of filamentous fungi and is a prerequisite for symbiotic and pathogenic host colonization achieved by molds. somatic cell-cell communication and subsequent cell fusion is governed by the mak-2 mitogen activated protein kinase (mapk) cascade in th ...201425411845
characterization of the neurospora crassa cell fusion proteins, ham-6, ham-7, ham-8, ham-9, ham-10, amph-1 and whi-2.intercellular communication of vegetative cells and their subsequent cell fusion is vital for different aspects of growth, fitness, and differentiation of filamentous fungi. cell fusion between germinating spores is important for early colony establishment, while hyphal fusion in the mature colony facilitates the movement of resources and organelles throughout an established colony. approximately 50 proteins have been shown to be important for somatic cell-cell communication and fusion in the mo ...201425279949
bioinformatic and biochemical characterizations of c-s bond formation and cleavage enzymes in the fungus neurospora crassa ergothioneine biosynthetic pathway.ergothioneine is a histidine thiol derivative. its mycobacterial biosynthetic pathway has five steps (egta-e catalysis) with two novel reactions: a mononuclear nonheme iron enzyme (egtb) catalyzed oxidative c-s bond formation and a plp-mediated c-s lyase (egte) reaction. our bioinformatic and biochemical analyses indicate that the fungus neurospora crassa has a more concise ergothioneine biosynthetic pathway because its nonheme iron enzyme, egt1, makes use of cysteine instead of γ-glu-cys as the ...201425275953
characterizing the roles of cryphonectria parasitica rna-dependent rna polymerase-like genes in antiviral defense, viral recombination and transposon transcript accumulation.an inducible rna-silencing pathway, involving a single dicer protein, dcl2, and a single argonaute protein, agl2, was recently shown to serve as an effective antiviral defense response in the chestnut blight fungus cryphonectria parasitica. eukaryotic rna-dependent rna polymerases (rdrps) are frequently involved in transcriptional and posttranscriptional gene silencing and antiviral defense. we report here the identification and characterization of four rdrp genes (rdr1-4) in the c. parasitica g ...201425268858
a protein kinase screen of neurospora crassa mutant strains reveals that the snf1 protein kinase promotes glycogen synthase phosphorylation.glycogen functions as a carbohydrate reserve in a variety of organisms and its metabolism is highly regulated. the activities of glycogen synthase and glycogen phosphorylase, the rate-limiting enzymes of the synthesis and degradation processes, respectively, are regulated by allosteric modulation and reversible phosphorylation. to identify the protein kinases affecting glycogen metabolism in neurospora crassa, we performed a screen of 84 serine/threonine kinase knockout strains. we identified mu ...201425253091
neurospora crassa as a model organism to explore the interconnected network of the cell cycle and the circadian clock.budding and fission yeast pioneered uncovering molecular mechanisms of eukaryotic cell division cycles. however, they do not possess canonical circadian clock machinery that regulates physiological processes with a period of about 24h. on the other hand, neurospora crassa played a critical role in elucidating molecular mechanisms of circadian rhythms, but have not been utilized frequently for cell cycle studies. recent findings demonstrate that there exists a conserved coupling between the cell ...201425239547
a high-throughput gene disruption methodology for the entomopathogenic fungus metarhizium robertsii.systematic gene disruption is a direct way to interrogate a fungal genome to functionally characterize the full suite of genes involved in various biological processes. metarhizium robertsii is extraordinarily versatile, and it is a pathogen of arthropods, a saprophyte and a beneficial colonizer of rhizospheres. thus, m. robertsii can be used as a representative to simultaneously study several major lifestyles that are not shared by the "model" fungi saccharomyces cerevisiae and neurospora crass ...201425222118
reaction-diffusion systems for spatio-temporal intracellular protein networks: a beginner's guide with two examples.spatio-temporal dynamics of a variety of proteins is, among other things, regulated by post-translational modifications of these proteins. such modifications can thus influence stability and biochemical activities of the proteins, activity and stability of their upstream targets within specific signalling pathways. commonly used mathematical tools for such protein-protein (and/or protein-mrna) interactions in single cells, namely, michaelis-menten and hill kinetics, yielding a system of ordinary ...201425210594
efficient gene targeting in ligase iv-deficient monascus ruber m7 by perturbing the non-homologous end joining pathway.inactivating the non-homologous end joining (nhej) pathway is a well established method to increase gene replacement frequency (grf) in filamentous fungi because nhej is predominant for the repair of dna double strand breaks (dsbs), while gene targeting is based on homologous recombination (hr). dna ligase iv, a component of the nhej system, is strictly required for the nhej in saccharomyces cerevisiae and neurospora crassa. to enhance the grf in monascus ruber m7, we deleted the mrlig4 gene enc ...201425209642
a family of starch-active polysaccharide monooxygenases.the recently discovered fungal and bacterial polysaccharide monooxygenases (pmos) are capable of oxidatively cleaving chitin, cellulose, and hemicelluloses that contain β(1→4) linkages between glucose or substituted glucose units. they are also known collectively as lytic pmos, or lpmos, and individually as aa9 (formerly gh61), aa10 (formerly cbm33), and aa11 enzymes. pmos share several conserved features, including a monocopper center coordinated by a bidentate n-terminal histidine residue and ...201425201969
regulated dna methylation and the circadian clock: implications in cancer.since the cloning and discovery of dna methyltransferases (dnmt), there has been a growing interest in dna methylation, its role as an epigenetic modification, how it is established and removed, along with the implications in development and disease. in recent years, it has become evident that dynamic dna methylation accompanies the circadian clock and is found at clock genes in neurospora, mice and cancer cells. the relationship among the circadian clock, cancer and dna methylation at clock gen ...201425198253
light-mediated participation of the vivid-like protein of fusarium fujikuroi vvda in pigmentation and development.the phytopathogen fusarium fujikuroi is a model fungus for the production of different secondary metabolites. these include the acidic xanthophyll neurosporaxanthin, produced through the activity of the enzymes encoded by the car genes, of which carra and carb play a major role. expression of these genes is induced by light but, in contrast to other fungi, the induction is not impaired under continuous illumination in null mutants of the f. fujikuroi wc-1-like gene, wcoa. therefore, we investiga ...201425154020
efficient detection of unpaired dna requires a member of the rad54-like family of homologous recombination proteins.meiotic silencing by unpaired dna (msud) is a process that detects unpaired regions between homologous chromosomes and silences them for the duration of sexual development. while the phenomenon of msud is well recognized, the process that detects unpaired dna is poorly understood. in this report, we provide two lines of evidence linking unpaired dna detection to a physical search for dna homology. first, we have found that a putative snf2-family protein (sad-6) is required for efficient msud in ...201425146971
vib1, a link between glucose signaling and carbon catabolite repression, is essential for plant cell wall degradation by neurospora crassa.filamentous fungi that thrive on plant biomass are the major producers of hydrolytic enzymes used to decompose lignocellulose for biofuel production. although induction of cellulases is regulated at the transcriptional level, how filamentous fungi sense and signal carbon-limited conditions to coordinate cell metabolism and regulate cellulolytic enzyme production is not well characterized. by screening a transcription factor deletion set in the filamentous fungus neurospora crassa for mutants una ...201425144221
global gene expression and focused knockout analysis reveals genes associated with fungal fruiting body development in neurospora crassa.fungi can serve as highly tractable models for understanding genetic basis of sexual development in multicellular organisms. applying a reverse-genetic approach to advance such a model, we used random and multitargeted primers to assay gene expression across perithecial development in neurospora crassa. we found that functionally unclassified proteins accounted for most upregulated genes, whereas downregulated genes were enriched for diverse functions. moreover, genes associated with development ...201424243796
a comparative systems analysis of polysaccharide-elicited responses in neurospora crassa reveals carbon source-specific cellular adaptations.filamentous fungi are powerful producers of hydrolytic enzymes for the deconstruction of plant cell wall polysaccharides. however, the central question of how these sugars are perceived in the context of the complex cell wall matrix remains largely elusive. to address this question in a systematic fashion we performed an extensive comparative systems analysis of how the model filamentous fungus neurospora crassa responds to the three main cell wall polysaccharides: pectin, hemicellulose and cell ...201424224966
solution structure and interface-driven self-assembly of nc2, a new member of the class ii hydrophobin proteins.hydrophobins are fungal proteins that self-assemble spontaneously to form amphipathic monolayers at hydrophobic:hydrophilic interfaces. hydrophobin assemblies facilitate fungal transitions between wet and dry environments and interactions with plant and animal hosts. nc2 is a previously uncharacterized hydrophobin from neurospora crassa. it is a highly surface active protein and is able to form protein layers on a water:air interface that stabilize air bubbles. on a hydrophobic substrate, nc2 fo ...201424218020
analysis of cellodextrin transporters from neurospora crassa in saccharomyces cerevisiae for cellobiose fermentation.saccharomyces cerevisiae can be engineered to ferment cellodextrins produced by cellulases as a product of cellulose hydrolysis. direct fermentation of cellodextrins instead of glucose is advantageous because glucose inhibits cellulase activity and represses the fermentation of non-glucose sugars present in cellulosic hydrolyzates. to facilitate cellodextrin utilization by s. cerevisiae, a fungal cellodextrin-utilizing pathway from neurospora crassa consisting of a cellodextrin transporter and a ...201424190499
molecular cloning and expression of fungal cellobiose transporters and β-glucosidases conferring efficient cellobiose fermentation in saccharomyces cerevisiae.cellobiose was once regarded as a byproduct that should be removed from biomass hydrolysates because of its inhibitory activity to cellulases. it was revealed, however, that cellobiose could serve as a co-substrate for xylose fermentation by engineered saccharomyces cerevisiae. despite its advantages, to date, little is known about cellodextrin transporters that endow s. cerevisiae with cellobiose transporting ability. in this study, engineered s. cerevisiae strains capable of fermenting cellobi ...201424184384
the gene cuta of fusarium fujikuroi, encoding a protein of the haloacid dehalogenase family, is involved in osmotic stress and glycerol metabolism.survival of micro-organisms in natural habitats depends on their ability to adapt to variations in osmotic conditions. we previously described the gene cut-1 of neurospora crassa, encoding a protein of the haloacid dehalogenase family with an unknown function in the osmotic stress response. here we report on the functional analysis of cuta, the orthologous gene in the phytopathogenic fungus fusarium fujikuroi. cuta mrna levels increased transiently after exposure to 0.68 m nacl and were reduced ...201424129845
retracted article: the effects of each beta-glucosidase gene deletion on cellulase gene regulation in neurospora crassa (online publication: doi 10.1007/s10482-013- 9972-7).takao kasuga and zhiliang fan were listed as co-authors without their acknowledgement and would like to be removed from the authors list. this article is retracted on request of weihua wu due to inconsistent errors of intracellular and total beta-glucosidase activities assay and the mycelia weight measurement in this paper. these errors led to inaccurate results of total and intracellular beta-glucosidase activities, and the normalized endoglucanases and exoglucanases activities and therefore co ...201424000093
interconnections of reactive oxygen species homeostasis and circadian rhythm in neurospora crassa.both circadian rhythm and the production of reactive oxygen species (ros) are fundamental features of aerobic eukaryotic cells. the circadian clock enhances the fitness of organisms by enabling them to anticipate cycling changes in the surroundings. ros generation in the cell is often altered in response to environmental changes, but oscillations in ros levels may also reflect endogenous metabolic fluctuations governed by the circadian clock. on the other hand, an effective regulation and timing ...201423964982
adoption of the q transcriptional regulatory system for zebrafish transgenesis.the gal4-uas regulatory system of yeast is widely used to modulate gene expression in drosophila; however, there are limitations to its usefulness in transgenic zebrafish, owing to progressive methylation and silencing of the cpg-rich multicopy upstream activation sequence. although a modified, less repetitive uas construct may overcome this problem, it is highly desirable to have additional transcriptional regulatory systems that can be applied independently or in combination with the gal4/uas ...201423792917
inorganic phosphate uptake in unicellular eukaryotes.inorganic phosphate (pi) is an essential nutrient for all organisms. the route of pi utilization begins with pi transport across the plasma membrane.201424674820
evolution of rna-protein interactions: non-specific binding led to rna splicing activity of fungal mitochondrial tyrosyl-trna synthetases.the neurospora crassa mitochondrial tyrosyl-trna synthetase (mttyrrs; cyt-18 protein) evolved a new function as a group i intron splicing factor by acquiring the ability to bind group i intron rnas and stabilize their catalytically active rna structure. previous studies showed: (i) cyt-18 binds group i introns by using both its n-terminal catalytic domain and flexibly attached c-terminal anticodon-binding domain (ctd); and (ii) the catalytic domain binds group i introns specifically via multiple ...201425536042
the aging biological clock in neurospora crassa.the biological clock affects aging through ras-1 (bd) and lag-1, and these two longevity genes together affect a clock phenotype and the clock oscillator in neurospora crassa. using an automated cell-counting technique for measuring conidial longevity, we show that the clock-associated genes lag-1 and ras-1 (bd) are true chronological longevity genes. for example, wild type (wt) has an estimated median life span of 24 days, while the double mutant lag-1, ras-1 (bd) has an estimated median life s ...201425535564
[lah-3, a transcriptional factor involved in osmotic regulation in neurospora crassa].[objective] in order to identify the function of lah-3 in osmotic regulation, we generated lah-3 deletion strain and analyzed its phenotype by osmostress treatment. [methods] we used homologous recombination to replace lah-3gene by hph gene and treated these cells with 4% nacl and 1 m sorbitol to analyze the phenotype. northern blot was used to detect the expressions of osmoresponsing genes. western blot was used to examine the phosphorylation level of lah-3 and os-2 and the expression of os-2. ...201425522587
selection and evaluation of reference genes for expression studies with quantitative pcr in the model fungus neurospora crassa under different environmental conditions in continuous culture.neurospora crassa has served as a model organism for studying circadian pathways and more recently has gained attention in the biofuel industry due to its enhanced capacity for cellulase production. however, in order to optimize n. crassa for biotechnological applications, metabolic pathways during growth under different environmental conditions must be addressed. reverse-transcription quantitative pcr (rt-qpcr) is a technique that provides a high-throughput platform from which to measure the ex ...201425474155
biomineralization of metal carbonates by neurospora crassa.in this research, the urease-positive fungus neurospora crassa was investigated for the biomineralization of calcium carbonate and its potential application in metal biorecovery and/or bioremediation. after 12 d incubation at 25 °c in urea and calcium-containing medium, extensive biomineralization of fungal filaments was observed. energy dispersive x-ray analysis of crystalline precipitates on the hyphae of n. crassa showed that the main elements present in the crystals were ca, c, and o. x-ray ...201425423300
endogenous ergothioneine is required for wild type levels of conidiogenesis and conidial survival but does not protect against 254 nm uv-induced mutagenesis or kill.ergothioneine, a histidine derivative, is concentrated in conidia of ascomycetous fungi. to investigate the function of ergothioneine, we crossed the wild type neurospora crassa (egt(+)) and an ergothioneine non-producer (egt(-), δegt-1, a knockout in ncu04343.5) and used the egt(+) and egt(-) progeny strains for phenotypic analyses. compared to the egt(+) strains, egt(-) strains had a 59% reduction in the number of conidia produced on vogel's agar. after storage of egt(+) and egt(-) conidia at ...201425446508
the histone h3 lysine 9 methyltransferase dim-5 modifies chromatin at frequency and represses light-activated gene expression.the transcriptional program controlling the circadian rhythm requires coordinated regulation of chromatin. characterization of the chromodomain helicase dna-binding enzyme chd1 revealed dna methylation in the promoter of the central clock gene frequency (frq) in neurospora crassa. in this report, we show that the dna methylation at frq is not only dependent on the dna methyltransferase dim-2 but also on the h3k9 methyltransferase dim-5 and hp1. histone h3 lysine 9 trimethylation (h3k9me3) occurs ...201425429045
xylitol production at high temperature by engineered kluyveromyces marxianus.several recombinant kluyveromyces marxianus strains were constructed through overexpressing the neurospora crassa xylose reductase genes. yzj015, which maintained the original xylitol dehydrogenase gene, produced xylitol with the highest productivity (1.49 g l(-1) h(-1)) from 100 g l(-1) xylose at 42 °c. even at 45 °c, yzj015 was still able to produce 60.03 g l(-1) xylitol from 100 g l(-1) xylose with a productivity of 1.25 g l(-1)h(-1). in addition, for 20 rounds of cell recycling at 42 °c, yzj ...201424291795
transcription factor ccg-8 as a new regulator in the adaptation to antifungal azole stress.antifungal azoles are widely used for controlling fungal infections. fungi are able to change the expression of many genes when they adapt to azole stress, and increased expression of some of these genes can elevate resistance to azoles. however, the regulatory mechanisms behind transcriptional adaption to azoles in filamentous fungi are poorly understood. in this study, we found that deletion of the transcription factor gene ccg-8, which is known to be a clock-controlled gene, made neurospora c ...201424342650
genetic and metabolomic dissection of the ergothioneine and selenoneine biosynthetic pathway in the fission yeast, s. pombe, and construction of an overproduction system.ergothioneine is a small, sulfur-containing metabolite (229 da) synthesized by various species of bacteria and fungi, which can accumulate to millimolar levels in tissues or cells (e.g. erythrocytes) of higher eukaryotes. it is commonly marketed as a dietary supplement due to its proposed protective and antioxidative functions. in this study we report the genes forming the two-step ergothioneine biosynthetic pathway in the fission yeast, schizosaccharomyces pombe. we identified the first gene, e ...201424828577
exploring microrna-like small rnas in the filamentous fungus fusarium oxysporum.rna silencing such as quelling and meiotic silencing by unpaired dna (msud) and several other classes of special small rnas have been discovered in filamentous fungi recently. more than four different mechanisms of microrna-like rnas (milrnas) production have been illustrated in the model fungus neurospora crassa including a dicer-independent pathway. to date, very little work focusing on small rnas in fungi has been reported and no universal or particular characteristic of milrnas were defined ...201425141304
the neurospora crassa cps-1 polysaccharide synthase functions in cell wall biosynthesis.the neurospora crassa cps-1 gene encodes a polysaccharide synthase with homology to the cryptococcus neoformans hyaluronic acid synthase cps1p. homologs of the cps-1 gene are found in the genomes of many fungi. loss of cps-1 results in a cell wall defect that affects all stages of the n. crassa life cycle, including vegetative growth, protoperithecia (female mating structure) development, and conidia (asexual spore) development. the cell wall of cps-1 deletion mutants is sensitive to cell wall p ...201424953997
long-read, whole-genome shotgun sequence data for five model organisms.single molecule, real-time (smrt) sequencing from pacific biosciences is increasingly used in many areas of biological research including de novo genome assembly, structural-variant identification, haplotype phasing, mrna isoform discovery, and base-modification analyses. high-quality, public datasets of smrt sequences can spur development of analytic tools that can accommodate unique characteristics of smrt data (long read lengths, lack of gc or amplification bias, and a random error profile le ...201425977796
regulation of vectorial supply of vesicles to the hyphal tip determines thigmotropism in neurospora crassa.thigmotropism is the ability of an organism to respond to a topographical stimulus by altering its axis of growth. the thigmotropic response of the model fungus neurospora crassa was quantified using microfabricated glass slides with ridges of defined height. we show that the polarity machinery at the hyphal tip plays a role in the thigmotropic response of n. crassa. deletion of n. crassa genes encoding the formin, bni-1, and the rho-gtpase, cdc-42, an activator of bni-1 in yeast, cdc-24, its gu ...201424607352
sensitivity of neurospora crassa to a marine-derived aspergillus tubingensis anhydride exhibiting antifungal activity that is mediated by the mas1 protein.the fungus aspergillus tubingensis (strain oy907) was isolated from the mediterranean marine sponge ircinia variabilis. extracellular extracts produced by this strain were found to inhibit the growth of several fungi. among the secreted extract components, a novel anhydride metabolite, tubingenoic anhydride a (1) as well as the known 2-carboxymethyl-3-hexylmaleic acid anhydride, asperic acid, and campyrone a and c were purified and their structure elucidated. compound 1 and 2-carboxymethyl-3-hex ...201425257783
neurospora crassa: looking back and looking forward at a model microbe.investigation of the red bread mold that contaminated french bakeries nearly two centuries ago has led to a wealth of discoveries that have impacted our understanding of genetic, biochemical, and molecular mechanisms in microbes, from mendelian genetics and the gene-enzyme relationship to circadian rhythm and plant cell wall degradation. early neurospora research focused on elucidating mechanisms of genetic recombination and gene action and later progressed to addressing complex biological quest ...201425480699
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
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
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
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
efficient formation of heterokaryotic sclerotia in the filamentous fungus aspergillus oryzae.heterokaryon formation by hyphal fusion occurs during a sexual/parasexual cycle in filamentous fungi, and therefore, it is biotechnologically important for crossbreeding. in the industrial filamentous fungus aspergillus oryzae, a parasexual cycle has been reported, and it was recently suggested that sexual reproduction should be possible. however, as a. oryzae enters into hyphal fusion with a much lower frequency than neurospora crassa, the process of heterokaryon formation has not been extensiv ...201424201891
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
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
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
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
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
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
purification and properties of neurospora crassa laccase.extracellular neurospora laccase (p-diphenol:oxygen oxidoreductase; ec 1.10.3.2) has been purified to apparent homogeneity by classical purification techniques. the enzyme, which consists of mainly one form, has a molecular weight of 64,800 and contains 11% carbohydrate. the ultraviolet, visible, and electron paramagnetic resonance spectra indicate that both type i and type ii copper are present, as described for the polyporus versicolor enzyme. with the exception of phloroglucinol, only para- a ...20144278681
genetic analysis of a suppressor of urease-deficiency in neurospora crassa. 20144281763
regulation of exocellular proteases in neurospora crassa: role of neurospora proteases in induction.cells of neurospora crassa strain 74a, grown on sucrose for 12 h and transferred to a medium containing protein as sole carbon source, would not produce exocellular protease in significant amounts. when a filtrate from a culture induced to make protease by normal growth on a medium containing protein as principal carbon source was added to an exponential-phase culture in protein medium, exocellular protease was made in amounts similar to those made during normal induction. the material in the cu ...20144270446
genetic recombination in neurospora crassa affected by changes in the supply of cellular energy. 20144281764
preparation of mitochondrial membrane proteins from neurospora crassa: prevention of lipid autoxidation damage by an antioxidant. 20144266161
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
cyanide-resistant respiration in neurospora crassa.cell respiration in wild type and poky was studied as part of a long-term investigation of cyanide-resistant respiration in neurospora. respiration in wild type proceeds via a cytochrome chain which is similar to that of higher organisms; it is sensitive to antimycin a or cyanide. poky, on the other hand, respires by means of two alternative oxidase systems. one of these is analogous to the wild-type cytochrome chain in that it can be inhibited by antimycin a or cyanide; this system accounts for ...20144333318
spin-labeled mitochondrial lipids in neurospora crassa. 20144301148
cold-induced increase of glycerol kinase activity in neurospora crassa: rapid inactivation of the enzyme in vivo.the glycerol kinase activity induced by incubation of neurospora crassa at low temperatures was rapidly lost when cultures were returned to 26 c. after a short lag, the activity disappeared irreversibly with a half-life of approximately 15 min. the loss of activity was not due to a change in the level of an inhibitor or activator. glycerol reduced the activity loss but did not completely prevent it, which was an effect that was dependent on protein synthesis. the cold-induced activity was also a ...20144281774
end-product regulation of the tryptophan-nicotinic acid pathway in neurospora crassa.the regulation of the tryptophan-nicotinic acid pathway in neurospora crassa was examined with mutants (nic-2, nic-3) which require nicotinamide for growth. the accumulation of n-acetylkynurenin and 3-hydroxyanthranilic acid by these mutants served to estimate the level of function of the early reactions in the pathway. in still cultures, maximal accumulation occurred with media containing growth-limiting amounts of nicotinamide; the accumulation of intermediates was almost negligible with nicot ...20144107105
thermostimulation of conidiation and succinic oxidative metabolism of neurospora crassa. 20144303684
regulation of a sulfur-controlled protease in neurospora crassa.wild-type neurospora crassa produces and secretes extracellular protease(s) when grown on a medium containing a protein as its principle sulfur source. readily available sulfur sources, such as sulfate or methionine, repress the synthesis of the proteolytic activity. preliminary characterization of the proteolytic enzyme shows it to have a molecular weight of about 31,000, a ph optimum of 6 to 9 with casein as substrate, and esterolytic activity against acetyl-tyrosine ethyl ester with a ph opti ...20144270448
interaction of galactosaminoglycan with neurospora conidia.the inactivation of neurospora crassa conidia by galactosaminoglycan isolated from cultures of this organism was followed by measuring colony-forming ability and ability to take up radiolabeled metabolites. when kinetic data on the loss of transport function and on killing were analyzed by use of target theory, it appeared that few "hits" are required for inactivation. however, studies with radio-labeled galactosaminoglycan mucopolysaccharides showed that cells receiving a single lethal hit have ...20144281775
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