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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
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
[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
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
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
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
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
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
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
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
biological activity of secondary metabolites isolated from mangrove fungi neurospora crassa.the screening of antimicrobial compounds from mangrove-associated fungi is a promising way to meet the increasing threat of drug-resistant strains of human pathogens. in the present study, a novel foliar fungus was isolated from the leaves of rhizophora mucronata. crude fungal extracts were obtained through solvent extraction. the fungal extracts exhibited an effective antimicrobial activity against bacterial strains. this study reports the cytotoxic activity of the secondary metabolite produced ...201324640249
heterologous expression and kinetic characterisation of neurospora crassa β-xylosidase in pichia pastoris.to degrade plant hemicelluloses fungi employ β-xylosidases to hydrolyse xylooligosaccharides, released by endo-xylanases, into xylose. we have expressed the β-xylosidase from neurospora crassa in pichia pastoris under the control of alcohol oxidase 1 (aox1) promoter. the recombinant enzyme is optimally active at 50 °c and ph 5.0 with km and vmax values of 8.9 mm and 1052 μmol min⁻¹ mg⁻¹ respectively against 4-nitrophenyl β-xylopyranoside. xylose is a non-competitive inhibitor with a k(i) of 1.72 ...201424629269
natural antisense transcripts and long non-coding rna in neurospora crassa.the prevalence of long non-coding rnas (lncrna) and natural antisense transcripts (nats) has been reported in a variety of organisms. while a consensus has yet to be reached on their global importance, an increasing number of examples have been shown to be functional, regulating gene expression at the transcriptional and post-transcriptional level. here, we use rna sequencing data from the abi solid platform to identify lncrna and nats obtained from samples of the filamentous fungus neurospora c ...201424621812
[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
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
evidence of a critical role for cellodextrin transporte 2 (cdt-2) in both cellulose and hemicellulose degradation and utilization in neurospora crassa.cdt-1 and cdt-2 are two cellodextrin transporters discovered in the filamentous fungus neurospora crassa. previous studies focused on characterizing the role of these transporters in only a few conditions, including cellulose degradation, and the function of these two transporters is not yet completely understood. in this study, we show that deletion of cdt-2, but not cdt-1, results in growth defects not only on avicel but also on xylan. cdt-2 can be highly induced by xylan, and this mutant has ...201424586693
transcriptional comparison of the filamentous fungus neurospora crassa growing on three major monosaccharides d-glucose, d-xylose and l-arabinose.d-glucose, d-xylose and l-arabinose are the three major monosaccharides in plant cell walls. complete utilization of all three sugars is still a bottleneck for second-generation cellulolytic bioethanol production, especially for l-arabinose. however, little is known about gene expression profiles during l-arabinose utilization in fungi and a comparison of the genome-wide fungal response to these three major monosaccharides has not yet been reported.201424581151
calcium/calmodulin-dependent kinases are involved in growth, thermotolerance, oxidative stress survival, and fertility in neurospora crassa.calcium/calmodulin-dependent kinases (ca(2+)/camks) are ser/thr protein kinases that respond to change in cytosolic free ca(2+) ([ca(2+)]c) and play multiple cellular roles in organisms ranging from fungi to humans. in the filamentous fungus neurospora crassa, four ca(2+)/cam-dependent kinases, ca(2+)/camk-1 to 4, are encoded by the genes ncu09123, ncu02283, ncu06177, and ncu09212, respectively. we found that camk-1 and camk-2 are essential for full fertility in n. crassa. the survival of ∆camk- ...201424570326
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
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
a new variant of self-excising β-recombinase/six cassette for repetitive gene deletion and homokaryon purification in neurospora crassa.in a previous study, we developed a cassette employing a bacterial β-recombinase acting on six recognition sequences (β-rec/six), which allowed repetitive site-specific gene deletion and marker recycling in neurospora crassa. however, only one positive selection marker was used in the cassette. a tedious subsequent procedure was needed to purify homokaryons due to the lack of a negative selection after cassette eviction. additionally, the endoxylanase xylp promoter from penicillium chrysogenum u ...201424556286
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
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
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
[light-dependent carotenoid synthesis : iv. the role of oxygen in photoinduction].the purpose of these studies was to investigate wether the primary step in the reaction chain of the light-dependent carotenoid synthesis in heterotrophic organisms, the photochemical reaction of the photoreceptor, requires oxygen. submerged cultures of fusarium aquaeductuum and neurospora crassa were used.biosynthesis of carotenoids following a photoinduction occurred only under aerobic conditions. when the mycelia were illuminated in the absence of oxygen and subsequently transferred to aerobi ...196824515321
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
bright to dim oscillatory response of the neurospora circadian oscillator.the fungus neurospora crassa constitutes an important model system extensively used in chronobiology. several studies have addressed how environmental cues, such as light, can reset or synchronize a circadian system. by means of an optimized firefly luciferase reporter gene and a controllable lighting system, we show that neurospora can display molecular circadian rhythms in dim light when cultures receive bright light prior to entering dim light conditions. we refer to this behavior as the "bri ...201424492882
a kinetic study of the effects of light on circadian rhythmicity of the frq promoter of neurospora crassa.the role of the frq gene in the neurospora crassa circadian rhythm has been widely studied, but technical limitations have hindered a thorough analysis of frq circadian expression waveform. through our experiments, we have shown an improved precision in defining neurospora's circadian rhythm kinetics using a codon optimized firefly luciferase gene reporter linked to a frq promoter. in vivo examination of this real-time reporter has allowed for a better understanding of the relationship of the li ...201424492881
[light dependent carotenoid synthesis : vii. cycloheximide or anaerobic conditions as reversible blocks of the sequence of light-induced reactions].anaerobic conditions completely block the synthesis of carotenoids in fusarium aquaeduciuum or neurospora crassa. even after sufficient illumination of the mycelia in the presence of air, the subsequent production of carotenoids in the dark is suppressed entirely when the fungi are transferred to an oxygen-free atmosphere. in turn, restoring aerobic conditions sets off the pigment synthesis in the dark without renewed photoinduction at any time within 48 hours, although the yields of carotenoids ...197124488430
photo morphogenesis and photo response of the blue-light receptor gene cmwc-1 in different strains of cordyceps militaris.light is a necessary environmental factor for stroma formation and development of cordyceps militaris, a well-known edible and medicinal fungus. in this study, photo morphogenesis and the blue-light receptor gene were studied using five representative strains of c. militaris. the results suggest that light was essential for colony pigmentation and could promote conidia production. cmwc-1, the homologe of the blue-light photoreceptor of neurospora crassa, was cloned from the genome of c. militari ...201424484244
circadian rhythms synchronize mitosis in neurospora crassa.the cell cycle and the circadian clock communicate with each other, resulting in circadian-gated cell division cycles. alterations in this network may lead to diseases such as cancer. therefore, it is critical to identify molecular components that connect these two oscillators. however, molecular mechanisms between the clock and the cell cycle remain largely unknown. a model filamentous fungus, neurospora crassa, is a multinucleate system used to elucidate molecular mechanisms of circadian rhyth ...201424474764
genetically engineered photoinducible homodimerization system with improved dimer-forming efficiency.vivid (vvd) is a photoreceptor derived from neurospora crassa that rapidly forms a homodimer in response to blue light. although vvd has several advantages over other photoreceptors as photoinducible homodimerization system, vvd has a critical limitation in its low dimer-forming efficiency. to overcome this limitation of wild-type vvd, here we conduct site-directed saturation mutagenesis in the homodimer interface of vvd. we have found that the ile52cys mutation of vvd (vvd-52c) substantially im ...201424428544
kinetic properties of de-repressible alkaline phosphatase (ec 3.1.3.1) from neurospora crassa.de-repressible alkaline phosphatase from n. crassa shows inhibition by pnp-p and a hyperbolic mixed-type inhibition by pi. both increasing concentrations of pi and decreases in assay ph abolished inhibition by the substrate. also, pi promoted polymerization of the enzyme molecule, whose effect may account for the inhibitory behaviour shown by the enzyme in the presence of low pi concentrations.199124424939
light-dependent carotenoid synthesis : x. lag-phase after a second illumination period in fusarium aquaeductuum and neurospora crassa.in dark grown mycelia of the fungi fusarium aquaeductuum lagh. and neurospora crassa the kinetics of carotenoid accumulation after brief illuminations were investigated. following a second illumination period photoinduced carotenoid biosynthesis started also only after a lag-phase. such a "secondary" lag-phase not only appeared when the pigment accumulation induced by the first light treatment had been completed, but also when the second illumination was given during the period of rapid caroteno ...197724420226
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
short communication: purification and properties of acid phosphatase (ec 3.1.3.2) secreted by strain 74a of the mould neurospora crassa.both pi-repressible acid phosphatases, iib (mycelial) and iic (extracellular), synthesized by neurospora crassa and purified to apparent homogeneity by 7.5% page, are monomers, are inhibited by 2 mm zncl2 and are non-specifically stimulated by salts. however, the iic form is activated by p-nitrophenylphosphate (in a negative co-operativity effect with a k 0.5 of 2.5 mm) whereas form iib shows michaelis kinetics, with a k m of 0.5 mm. thus, since both enzymatic forms may be expressed by the same ...199624415104
purification of neurospora crassa alkaline phosphatase without dnase activity for use in molecular biology.alkaline phosphatase, excreted by neurospora crassa preg (c) and purified to apparent homogeneity by 7.5% page, did not show dnaase activity and removed the terminal 5'-phosphate group from plasmid bluescript m13(+) linearized with ecori. the preg (c) strain may therefore replace other sources of alkaline phosphatase for use in dephosphorylating linearized plasmidial dna in cloning experiments.199524414902
circadian rhythms in neurospora crassa on a polydimethylsiloxane microfluidic device for real-time gas perturbations.racetubes, a conventional system employing hollow glass tubes, are typically used for monitoring circadian rhythms from the model filamentous fungus, neurospora crassa. however, a major technical limitation in using a conventional system is that racetubes are not amenable for real-time gas perturbations. in this work, we demonstrate a simple microfluidic device combined with real-time gas perturbations for monitoring circadian rhythms in neurospora crassa using bioluminescence assays. the develo ...201324404062
assessing the effects of light on differentiation and virulence of the plant pathogen botrytis cinerea: characterization of the white collar complex.organisms are exposed to a tough environment, where acute daily challenges, like light, can strongly affect several aspects of an individual's physiology, including pathogenesis. while several fungal models have been widely employed to understand the physiological and molecular events associated with light perception, various other agricultural-relevant fungi still remain, in terms of their responsiveness to light, in the dark. the fungus botrytis cinerea is an aggressive pathogen able to cause ...201324391918
distribution and evolution of het gene homologs in the basidiomycota.in filamentous fungi a system known as somatic incompatibility (si) governs self/non-self recognition. si is controlled by a regulatory signaling network involving proteins encoded at the het (heterokaryon incompatible) loci. despite the wide occurrence of si, the molecular identity and structure of only a small number of het genes and their products have been characterized in the model fungi neurospora crassa and podospora anserina. our aim was to identify and study the distribution and evoluti ...201424380733
the importance of pyroglutamate in cellulase cel7a.the commercialization of lignocellulosic biofuels relies in part on the ability to engineer cellulase enzymes to have properties compatible with practical processing conditions. the cellulase cel7a has been a common engineering target because it is present in very high concentrations in commercial cellulase cocktails. significant effort has thus been focused on its recombinant expression. in particular, the yeast saccharomyces cerevisiae has often been used both in the engineering and basic stud ...201424375151
fungal gene expression levels do not display a common mode of distribution.rna-seq studies in metazoa have revealed a distinct, double-peaked (bimodal) distribution of gene expression independent of species and cell type. however, two studies in filamentous fungi yielded conflicting results, with a bimodal distribution in pyronema confluens and varying distributions in sordaria macrospora. to obtain a broader overview of global gene expression distributions in fungi, an additional 60 publicly available rna-seq data sets from six ascomycetes and one basidiomycete were a ...201324373411
neurospora crassa ncs-1, mid-1 and nca-2 double-mutant phenotypes suggest diverse interaction among three ca(2+)-regulating gene products. 201324371176
molecular analysis of intragenic recombination at the tryptophan synthetase locus in neurospora crassa.fifteen different classically generated and mapped mutations at the tryptophan synthetase locus in neurospora crassa have been characterized to the level of the primary sequence of the gene. this sequence analysis has demonstrated that intragenic recombination is accurate to order mutations within one open reading frame. while classic genetic analysis correctly ordered the mutations, the position of mutations characterized by gene sequence analysis was more accurate. a leaky mutation was found t ...201324371173
determinants of regioselective hydroxylation in the fungal polysaccharide monooxygenases.the ubiquitous fungal polysaccharide monooxygenases (pmos) (also known as gh61 proteins, lpmos, and aa9 proteins) are structurally related but have significant variation in sequence. a heterologous expression method in neurospora crassa was developed as a step toward connecting regioselectivity of the chemistry to pmo phylogeny. activity assays, as well as sequence and phylogenetic analyses, showed that the majority of fungal pmos fall into three major groups with distinctive active site surface ...201424350607
global analysis of serine/threonine and tyrosine protein phosphatase catalytic subunit genes in neurospora crassa reveals interplay between phosphatases and the p38 mitogen-activated protein kinase.protein phosphatases are integral components of the cellular signaling machinery in eukaryotes, regulating diverse aspects of growth and development. the genome of the filamentous fungus and model organism neurospora crassa encodes catalytic subunits for 30 protein phosphatase genes. in this study, we have characterized 24 viable n. crassa phosphatase catalytic subunit knockout mutants for phenotypes during growth, asexual development, and sexual development. we found that 91% of the mutants had ...201424347630
evidence for transceptor function of cellodextrin transporters in neurospora crassa.neurospora crassa colonizes burnt grasslands and metabolizes both cellulose and hemicellulose from plant cell walls. when switched from a favored carbon source to cellulose, n. crassa dramatically up-regulates expression and secretion of genes encoding lignocellulolytic enzymes. however, the means by which n. crassa and other filamentous fungi sense the presence of cellulose in the environment remains unclear. previously, we have shown that a n. crassa mutant carrying deletions of three β-glucos ...201424344125
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
a c4-oxidizing lytic polysaccharide monooxygenase cleaving both cellulose and cello-oligosaccharides.lignocellulosic biomass is a renewable resource that significantly can substitute fossil resources for the production of fuels, chemicals, and materials. efficient saccharification of this biomass to fermentable sugars will be a key technology in future biorefineries. traditionally, saccharification was thought to be accomplished by mixtures of hydrolytic enzymes. however, recently it has been shown that lytic polysaccharide monooxygenases (lpmos) contribute to this process by catalyzing oxidati ...201424324265
conserved rna helicase frh acts nonenzymatically to support the intrinsically disordered neurospora clock protein frq.protein conformation dictates a great deal of protein function. a class of naturally unstructured proteins, termed intrinsically disordered proteins (idps), demonstrates that flexibility in structure can be as important mechanistically as rigid structure. at the core of the circadian transcription/translation feedback loop in neurospora crassa is the protein frequency (frq), shown here shown to share many characteristics of idps. frq in turn binds to frequency-interacting rna helicase (frh), who ...201324316221
the biphasic fluence response of carotenogenesis in neurospora crassa: temporary insensitivity of the photoreceptor system.whether or not illumination is continuous or interrupted during the span in which increasing illumination time periods (i.e., increasing fluences) have no different effect on carotenogenesis is optional, in regard to the amount of carotenoids produced (revealing the plateau of the biphasic fluence response curve). this indicates temporary insensitivity. when the time delay between the onsets of the initial and second illumination is extended beyond the expanse of the plateau, the amount of carot ...198124301981
the ontogeny of lipid bodies (spherosomes) in plant cells : ultrastructural evidence.maturing embryos of 16 oil plants, anise suspension culture cells, and neurospora crassa cells were prepared for electron microscopy at different stages during massive lipid accumulation. lipid-rich structures of certain species were best preserved by dehydration of fixed tissues in ethanol without propylene oxide, embedding in spurr's medium, and polymerization at room temperature. in all cells examined, spherical lipid bodies (spherosomes) showed a moderately osmiophilic, amorphous matrix and ...198124301718
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
occurrence of xanthine dehydrogenase in chlamydomonas reinhardii: a common cofactor shared by xanthine dehydrogenase and nitrate reductase.wild-type chlamydomonas reinhardii cells have xanthine dehydrogenase activity when grown with nitrate, nitrite, urea, or amino acid media. mutant strains 102, 104, and 307 of chlamydomonas, lacking both xanthine dehydrogenase and nitrate reductase activities, were incapable of restoring the nadph-nitrate reductase activity of the mutant nit-1 of neurospora crassa, whereas wild type cells and mutants 203 and 305 had xanthine dehydrogenase and were able to reconstitute the nitrate reductase activi ...198124276829
light-induced phase shifting of the circadian clock in neurospora crassa requires ammonium salts at high ph.effects of external ionic conditions on light induced phase shifting of the circadian rhythm of conidiation in neurospora crassa were examined in simple buffer solutions for discerning effects of individual ions. mycelia were cultured to liquid media of different phs and then transferted to 10 mm piperazine-n,n'-bis(2-ethanesulmonic acid) (pipes) buffer of various phs and irradiated with while light. the phase of the rhythm of dark controls was not changed by transfer from medium to buffer. when ...198224271975
behaviour of dna-induced inositol-independent transformants of neurospora crassa in sexual crosses.treatment of inositolless (inl) strains of neurospora crassa with dna from the wild type (allo-dna) gives rise to inositol-independent (inl (+)) colonies. some of these dna-induced inl (+) strains (transformants) are sterile in sexual crosses on minimal medium that selects for the maintaining of the inl (+) character. the same inl (+) transformants, when crossed with an inl standard strain, are fertile on complete (inositol-containing) medium. there are, however, an increased number of unusual n ...198224270340
neurospora crassa protein arginine methyl transferases are involved in growth and development and interact with the ndr kinase cot1.the protein arginine methyltransferaseas (prmts) family is conserved from yeast to human, and regulates stability, localization and activity of proteins. we have characterized deletion strains corresponding to genes encoding for prmt1/3/5 (designated amt-1, amt-3 and skb-1, respectively) in neurospora crassa. deletion of prmt-encoding genes conferred altered arg-methylated protein profiles, as determined immunologically. δamt-1 exhibited reduced hyphal elongation rates (70% of wild type) and inc ...201324260473
effect of 2,4,5-trichlorophenol on hyphal membrane potentials in rhythmic mutants of neurospora crassa.the uncoupler 2,4,5-trichlorophenol (tcp) was used to test for differences in maintaining the hyphal membrane potentials in the wild type and rhythmic mutants of neurospora crassa. trichlorophenol (0.1 mmol·1(-1)) resulted in a depolarization of 93 mv (wild type) and 144 mv in the mutant clock. a total recovery was achieved in both strains after washing out the uncoupler. the circadian conidiation mutant band was more sensitive than the two other strains and showed two different reaction pattern ...198224257777
human-like eukaryotic translation initiation factor 3 from neurospora crassa.eukaryotic translation initiation factor 3 (eif3) is a key regulator of translation initiation, but its in vivo assembly and molecular functions remain unclear. here we show that eif3 from neurospora crassa is structurally and compositionally similar to human eif3. n. crassa eif3 forms a stable 12-subunit complex linked genetically and biochemically to the 13(th) subunit, eif3j, which in humans modulates mrna start codon selection. based on n. crassa genetic analysis, most subunits in eif3 are e ...201324250809
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
are carotenoids the blue-light photoreceptor in the photoinduction of protoperithecia in neurospora crassa?a triple albino mutant of neurospora crassa with a measured content of carotenoids absorbing at 470 nm less than 0.5% of that of the wild type (calculated value less than 8·10(-4)%) had the same threshold for photoinduction of protoperithecia as the wild type when illuminated with monochromatic light at 471 nm. this is strong evidence against the hypothesis that the bulk of carotenoids are the blue-light photoreceptor for this phenomenon. however, it is impossible to exclude traces of carotenoid ...198624233735
role of nitrogen in the photoinduction of protoperithecia and carotenoids in neurospora crassa.nitrogen, as kno3 or nh4no3, can inhibit the photoinduction of protoperithecia in neurospora crassa when present in the medium at a high concentration but does not inhibit the photoinduction of carotenoids. the point at which the presence of high nitrogen levels is no longer inhibitory is 5 h after illumination.198724232879
wheat germ agglutinin in wheat seedling roots: induction by elicitors and fungi.treatment of wheat (triticum aestivum l.) seedlings with elicitors originating from either plant or fungal cell walls induces about a 2-fold increase of wheat germ agglutinin (wga) in the roots. while the wga content in roots of healthy plants normally decreases as a function of germination time, a transient accumulation of wga could be observed in plants challenged with different fungi, including rhizoctonia solani, fusarium culmorum, pythium ultimum and neurospora crassa. peak levels in challe ...199024226823
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
phospho-regulation of the neurospora crassa septation initiation network.proper cell division is essential for growth and development of uni- and multicellular organisms. the fungal septation initiation network (sin) functions as kinase cascade that connects cell cycle progression with the initiation of cytokinesis. miss-regulation of the homologous hippo pathway in animals results in excessive cell proliferation and formation of tumors, underscoring the conservation of both pathways. how sin proteins interact and transmit signals through the cascade is only beginnin ...201324205386
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
temperature-sensitive nuclear mutants of neurospora crassa deficient in mitochondrial ribosomes.we have isolated two non-allelic nuclear mutants of neurospora crassa that are temperature-sensitive for the production of cytochromes aa 3 and b. when grown at 23 °c the mutants are virtually indistinguishable from the parent wild-type strains. when grown at 41 °c the mutants have large amounts of kcn-insensitive respiration and lack cytochromes aa 3 and b. further examination of the mutants revealed that they were extremely deficient in their capacity for mitochondrial protein synthesis when g ...197924190802
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
high frequency transfer of species specific mitochondrial dna sequences between members of the aspergillaceae.the mitochondrial genome of aspergillus nidulans var. echinulatus is approximately 20% larger than that of the closely related species aspergillus nidulans (eidam) winter. restriction enzyme mapping and electron microscopy has revealed that the size difference is due to the presence of six inserted sequences in the former. with the exception of a small number of species specific restriction sites and the six insertions/deletions, the two mitochondrial genomes appear identical. protoplast fusion ...198124190134
a "stopper" mutant of neurospora crassa containing two populations of aberrant mitochondrial dna.[e35], an extranuclear mutant of neurospora crassa has all the phenotypic characteristics of the "stopper" mutants (de vries et al. 1980). in the present work, the mitochondrial dna as well as the mitochondrial translation products are characterized further. the primary mutational event appears to have been the deletion of about 4 kbp from the wild-type genome. moreover, after prolonged vegetative growth the mutant accumulates an 8-μm circular mtdna, which was demonstrated both by electronmicros ...198124190132
a regulatory phenotype associated with the en-am 1 mutant of neurospora crassa.we have investigated the nature of the en-am1 mutant of neurospora crassa and have found that it affects the regulation of proline oxidase and utilisation of other nitrogen sources. this mutant is closely linked to the gln gene but not allelic with it. data from crosses suggest that the two genes he on opposite sides of the in1 gene on linkage group vr.198224186376
genetic map of mitochondrial dna in podospora anserina.in order to develop an eukaryotic vector with the podospora plasmid, further characterization is required of the mitochondrial dna into which this plasmid is integrated, a physical map (restriction sites) of the podospora chondriome (size 95 kb) has been completed. as prerequisite for the establishment of a genetic (functional) map, 70% of the chondriome was cloned in e. coli vectors. using mitochondrial genes from saccharomyces cerevisiae, six structural genes were located on the podospora chon ...198224186230
characterization of a variant neurospora crassa mitochondrial dna which contains tandem reiterations of a 1.9 kb sequence.we have identified a second variant of neurospora mtdna which contains tandem, head-to-tail repeats of sequences at the boundary of eco ri-4 and 6. this region may contain a. major replication origin of neurospora mtdna.198124185958
cold-sensitive mutation in neurospora crassa affecting the production of 17s ribosomal rna from ribosomal precursor rna.the cold-sensitive mutant strain of neurospora crassa, crib-1 (pj30201) has a conditional defect in ribosome biosynthesis. at 10 °c this mutant underproduces the small (37s) ribosomal subunit. experiments were done to study rrna synthesis in crib-1 after a shift from the permissive (25 °c) to the nonpermissive temperature and vice versa. the results showed that the primary cold-sensitive defect in crib-1 is in the function, rather than the synthesis, of a molecule needed for the production of th ...198124185860
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
mitochondrial dna of the filamentous ascomycete cochliobolus heterostrophus : characterization of the mitochondrial chromosome and population genetics of a restriction enzyme polymorphism.the mitochondrial chromosome of cochliobolus heterostrophus is a circle approximately 115 kb in circumference, among the largest known from fungi. a physical map of c. heterostrophus mtdna was constructed using the restriction enzymes bamhi, ecori, and pvuli by dna-dna hybridizations with cloned or purified mtdna bamhi fragments. the following sequences were located on the mtdna map: (1) the large and small mitochondrial ribosomal rna genes (identified by heterologous hybridization to cloned neu ...198424178002
cloning and preliminary characterization of a molybdenum cofactor gene of neurospora crassa.a neurospora crassa library, constructed in a derivative of the plasmid pbr322 (prk9), was used to transform two e. coli ch1d molybdenum cofactor mutants (ch1d, ch1d::mu). subsequently, one transformant from each of three independent transformation experiments was restriction mapped. all three transformants had an identical n. crassa dna insert (4.2 kb). southern blot analysis with one of the plasmids (pmoco, 1:4) showed hybridization to a single band of n. crassa genomic dna. when pmoco plasmid ...198424177998
biochemical characterization of the molybdenum cofactor mutants of neurospora crassa: in vivo and in vitro reconstitution of nadph-nitrate reductase activity.molybdenum cofactor (moco) mutants of neurospora crassa lack both nadph-nitrate reductase and xanthine dehydrogenase activity. in vivo and in vitro studies to further characterize these mutants are now reported. the moco mutants nit-9a and nit-9b are capable of growing, albeit poorly, with nitrate as the sole nitrogen source, provided high levels of molybdate are present. the moco mutants nit-9a, nit-9b and nit-9c, but not nit-1, nit-7 or nit-8, have significant levels of nadph-nitrate reductase ...198424177997
a mitochondrial reading frame which may code for a second form of atpase subunit 9 in aspergillus nidulans.the nucleotide sequence of a 74 codon reading frame from the aspergillus nidulans mitochondrial genome is presented. the derived amino acid sequence displays typical features of dicyclohexylcarbodiimide (dccd) binding proteins and is 84% homologous with a mitochondrial reading frame that potentially encodes an atpase subunit 9 polypeptide in neurospora crassa. however, in a. nidulans, as in n. crassa, there is strong biochemical and genetic evidence that this subunit is in fact nuclearly-encoded ...198424177948
isolation and characterization of a methylammonium resistant mutant of neurospora crassa.a mutant of neurospora crassa has been isolated which is resistant to methylammonium, a structural analog of ammonium. in contrast to wild type, this mutant, mea-1, has derepressed nitrate reductase and nitrite reductase activities in the presence of ammonium. however, glutamine still represses these nitrate assimilation enzymes in mea-1. the nit-2 mutant was epistatic to mea-1 since the mea-1; nit-2 double mutant has the nit-2 mutant phenotype. in addition, mea-1; nit-2 double mutants cannot ut ...198424177912
structure and function of the trp3 gene of saccharomyces cerevisiae: analysis of transcription, promoter sequence, and sequence coding for a glutamine amidotransferase.the structure and function of the trp3 gene of saccharomyces cerevisiae were analyzed. subcloning of an original 4.8 kb bamhi dna fragment, carrying the yeast trp3 gene, allowed for a localization of the gene on a 2.5 kb clai/bamhi fragment. transcription was found to proceed from the clai site towards the bamhi site. three major transcription start sites were determined at positions -92, -87, and -81 by s1-mapping. the synthesis of the trp3 gene is regulated by the general control, and was foun ...198424177735
a new, rapid and efficient transformation procedure for neurospora.a new, rapid, efficient and reliable method for transforming neurospora crassa is described. in this procedure, germinated conidia are treated with lithium acetate, then incubated with dna, followed by exposure to polyethylene glycol and then a brief heat shock, prior to plating on selective medium. optimal conditions to achieve a high transformation rate are reported. transformation can be obtained with both circular and linear plasmid dna and also with genomic dna. although the rate is substan ...198424177533
rnai function, diversity, and loss in the fungal kingdom.rnai is conserved and has been studied in a broad cross-section of the fungal kingdom, including neurospora crassa, schizosaccharomyces pombe, cryptococcus neoformans, and mucor circinelloides. and yet well known species, including the model yeast saccharomyces cerevisiae and the plant pathogen ustilago maydis, have lost rnai, providing insights and opportunities to illuminate benefits conferred both by the presence of rnai and its loss. some of the earliest studies of rnai were conducted in neu ...201324173579
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