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
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 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
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
a new variant of self-excising β-recombinase/six cassette for repetitive gene deletion and homokaryon purification in neurospora 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
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
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
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
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
distribution and evolution of het gene homologs in the 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
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
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
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
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
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
location and contribution of individual β-glucosidase from neurospora crassa to total β-glucosidase activity.this study investigated the cellular location and the contribution of individual β-glucosidase (bgl) to total bgl activity in neurospora crassa. among the seven bgl genes, bgl3, bgl5, and bgl7 were transcribed at basal levels, whereas bgl1, bgl2, bgl4, and bgl6 were significantly up-regulated when the wild-type strain was induced with cellulose (avicel). bgl1 and bgl4 were found to be contributors to intracellular bgl activity, whereas the activities of bgl2 and bgl6 were mainly extracellular. s ...201324162785
regulation of gene expression in neurospora crassa with a copper responsive promoter.precise control of gene expression is a powerful method to elucidate biological function, and protein overexpression is an important tool for industry and biochemistry. expression of the neurospora crassa tcu-1 gene (ncu00830), encoding a high-affinity copper transporter, is tightly controlled by copper availability. excess copper represses, and copper depletion, via the use of a copper chelator, activates expression. the kinetics of induction and repression of tcu-1 are rapid, and the effects a ...201324142928
polygalacturonases from moniliophthora perniciosa are regulated by fermentable carbon sources and possible post-translational modifications.we report the first molecular and in silico analysis of monilophthora perniciosa polygalacturonases (pgs). three mppg genes (mppg1, mppg2 and mppg3) were identified and analyzed at transcriptional level, by rt-qpcr, in dikaryotic m. perniciosa mycelium grown on solid-bran based medium and on liquid medium supplemented with different fermentable and non-fermentable carbon sources. the mppg genes presented different expression patterns suggesting different individual regulation. however, all are m ...201324140149
sers properties of different sized and shaped gold nanoparticles biosynthesized under different environmental conditions by neurospora crassa extract.surface-enhanced raman scattering (sers) is a surface-sensitive technique that enhances raman scattering by molecules adsorbed on rough metal surfaces. it is known that metal nanoparticles, especially gold and silver nanoparticles, exhibit great sers properties, which make them very attractive for the development of biosensors and biocatalysts. on the other hand, the development of ecofriendly methods for the synthesis of metallic nanostructures has become the focus of research in several countr ...201324130891
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
neurospora crassa, a model system for epigenetics research.the filamentous fungus neurospora crassa has provided a rich source of knowledge on epigenetic phenomena that would have been difficult or impossible to gain from other systems. neurospora sports features found in higher eukaryotes but absent in both budding and fission yeast, including dna methylation and h3k27 methylation, and also has distinct rna interference (rnai)-based silencing mechanisms operating in mitotic and meiotic cells. this has provided an unexpected wealth of information on gen ...201324086046
discovery of cellobionic acid phosphorylase in cellulolytic bacteria and fungi.a novel phosphorylase was characterized as new member of glycoside hydrolase family 94 from the cellulolytic bacterium xanthomonas campestris and the fungus neurospora crassa. the enzyme catalyzed reversible phosphorolysis of cellobionic acid. we propose 4-o-β-d-glucopyranosyl-d-gluconic acid: phosphate α-d-glucosyltransferase as the systematic name and cellobionic acid phosphorylase as the short names for the novel enzyme. several cellulolytic fungi of the phylum ascomycota also possess homolog ...201324055472
convergent transcription induces dynamic dna methylation at disirna loci.cytosine methylation of dna is an important epigenetic gene silencing mechanism in plants, fungi, and animals. in the filamentous fungus neurospora crassa, nearly all known dna methylations occur in transposon relics and repetitive sequences, and dna methylation does not depend on the canonical rnai pathway. disirnas are dicer-independent small non-coding rnas that arise from gene-rich part of the neurospora genome. here we describe a new type of dna methylation that is associated with the disir ...201324039604
early colony establishment in neurospora crassa requires a map kinase regulatory network.vegetative fusion is essential for the development of an interconnected colony in many filamentous fungi. in the ascomycete fungus neurospora crassa, vegetative fusion occurs between germinated conidia (germlings) via specialized structures termed "conidial anastomosis tubes" (cats) and between hyphae within a mature colony. in n. crassa, both cat and hyphal fusion are under the regulation of a conserved map kinase cascade (nrc1, mek2, and mak2). here we show that the predicted downstream target ...201324037267
ham-2 and ham-3 are central for the assembly of the neurospora stripak complex at the nuclear envelope and regulate nuclear accumulation of the map kinase mak-1 in a mak-2-dependent manner.intercellular communication and somatic cell fusion are important for fungal colony establishment, multicellular differentiation and have been associated with host colonization and virulence of pathogenic species. by a combination of genetic, biochemical and live cell imaging techniques, we characterized the neurospora crassa stripak complex that is essential for self-signalling and consists of the six proteins ham-2/strip, ham-3/striatin, ham-4/slmap, mob-3/phocein, ppg-1/pp2a-c and pp2a-a. we ...201324028079
coexistence and expression profiles of two alternative splice variants of the pheromone receptor gene pre‑1 in neurospora this study, we show that two splice variants of the pheromone receptor gene (pre-1) transcript coexist in vegetative and reproductive tissues of the filamentous ascomycete fungus neurospora crassa. the two splice variants differ by intron retention of the last intron, which is predicted to result in a premature stop codon and loss of 322 amino acids in the c-terminal cytosolic region of pre-1. using quantitative pcr, we show that expression of the variants is influenced by mating type (mat), ...201324022114
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
is the fungus magnaporthe losing dna methylation?the long terminal repeat retrotransposon, magnaporthe gypsy-like element (maggy), has been shown to be targeted for cytosine methylation in a subset of magnaporthe oryzae field isolates. analysis of the f1 progeny from a genetic cross between methylation-proficient (br48) and methylation-deficient (gfsi1-7-2) isolates revealed that methylation of the maggy element was governed by a single dominant gene. positional cloning followed by gene disruption and complementation experiments revealed that ...201323979580
mass flow and velocity profiles in neurospora hyphae: partial plug flow dominates intra-hyphal transport.movement of nuclei, mitochondria and vacuoles through hyphal trunks of neurospora crassa were vector-mapped using fluorescent markers and green fluorescent protein tags. the vectorial movements of all three were strongly correlated, indicating the central role of mass (bulk) flow in cytoplasm movements in n. crassa. profiles of velocity versus distance from the hyphal wall did not match the parabolic shape predicted by the ideal hagen-poiseuille model of flow at low reynolds number. instead, the ...201323970568
evolutionary dynamics of sex-biased genes in a hermaphrodite fungus.differential gene expression is believed to largely explain sexually dichotomous phenotypes. this phenomenon is especially significant in hermaphrodites, in which male and female sexual tissues have identical genotypes. sex differences in transcription have been linked to molecular evolution: genes with higher expression in male compared with female sexual tissues (i.e., male-biased genes) have been associated with rapid gene divergence in various animals and plants, implying that selective diff ...201323966547
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
the n-terminal region of the neurospora ndr kinase cot1 regulates morphology via its interactions with mob2a/b.nuclear dbf2p-related (ndr) protein kinases are important for cell differentiation and polar morphogenesis in various organisms, yet some of their functions are still elusive. dysfunction of the neurospora crassa ndr kinase cot1 leads to cessation of tip extension and hyperbranching. ndr kinases require the physical interaction between the kinase's n-terminal region (ntr) and the mps1-binding (mob) proteins for their activity and functions. to study the interactions between cot1 and mob2 protein ...201323962317
investigation of the functional role of aldose 1-epimerase in engineered cellobiose utilization.functional expression of a cellodextrin transporter and an intracellular β-glucosidase from neurospora crassa in saccharomyces cerevisiae enables simultaneous co-fermentation of cellobiose and non-glucose sugars such as xylose. here we investigate the functional role of aldose 1-epimerase (aep) in engineered cellobiose utilization. one aep (gal10) and two putative aeps (yhr210c and ynr071c sharing 50.6% and 51.0% amino acid identity with gal10, respectively) were selected. deletion of gal10 led ...201323954547
mtb-3, a microtubule plus-end tracking protein (+tip) of neurospora crassa.the microtubule (mt) "plus end" constitutes the platform for the accumulation of a structurally and functionally diverse group of proteins, collectively called "mt plus-end tracking proteins" (+tips). +tips control mt dynamics and link mts to diverse sub-cellular structures. neurospora crassamicrotubule binding protein-3 (mtb-3) is the homolog of yeast eb1, a highly conserved +tip. to address the function of mtb-3, we examined strains with mtb-3 deletions, and we tagged mtb-3 with gfp to assess ...201323950979
analysis of mutations in neurospora crassa ermes components reveals specific functions related to β-barrel protein assembly and maintenance of mitochondrial morphology.the endoplasmic reticulum mitochondria encounter structure (ermes) tethers the er to mitochondria and contains four structural components: mmm1, mdm12, mdm10, and mmm2 (mdm34). the gem1 protein may play a role in regulating ermes function. saccharomyces cerevisiae and neurospora crassa strains lacking any of mmm1, mdm12, or mdm10 are known to show a variety of phenotypic defects including altered mitochondrial morphology and defects in the assembly of β-barrel proteins into the mitochondrial out ...201323940790
genome wide association identifies novel loci involved in fungal communication.understanding how genomes encode complex cellular and organismal behaviors has become the outstanding challenge of modern genetics. unlike classical screening methods, analysis of genetic variation that occurs naturally in wild populations can enable rapid, genome-scale mapping of genotype to phenotype with a medium-throughput experimental design. here we describe the results of the first genome-wide association study (gwas) used to identify novel loci underlying trait variation in a microbial e ...201323935534
reconstruction and validation of a genome-scale metabolic model for the filamentous fungus neurospora crassa using farm.the filamentous fungus neurospora crassa played a central role in the development of twentieth-century genetics, biochemistry and molecular biology, and continues to serve as a model organism for eukaryotic biology. here, we have reconstructed a genome-scale model of its metabolism. this model consists of 836 metabolic genes, 257 pathways, 6 cellular compartments, and is supported by extensive manual curation of 491 literature citations. to aid our reconstruction, we developed three optimization ...201323935467
characterization of bud emergence 46 (bem46) protein: sequence, structural, phylogenetic and subcellular localization analyses.the bud emergence 46 (bem46) protein from neurospora crassa belongs to the α/β-hydrolase superfamily. recently, we have reported that the bem46 protein is localized in the perinuclear er and also forms spots close by the plasma membrane. the protein appears to be required for cell type-specific polarity formation in n. crassa. furthermore, initial studies suggested that the bem46 amino acid sequence is conserved in eukaryotes and is considered to be one of the widespread conserved "known unknown ...201323916612
recent advances in septum biogenesis in neurospora crassa.hyphae of the ascomycota are tubular cells compartmentalized by perforated septa, whose central pore allows the flow of organelles and cytoplasm. while in plants and yeast septation leads to cell separation, in filamentous fungi the formation of crosswalls appears to have an architectural role, limits the extent of mechanical damage thus maintaining hyphal integrity, and also is of fundamental importance as part of cell differentiation. the increasing number of available fungal genome sequences, ...201323890213
synchronous activation of cell division by light or temperature stimuli in the dimorphic yeast schizosaccharomyces japonicus.many fungi respond to light and regulate fungal development and behavior. a blue light-activated complex has been identified in neurospora crassa as the product of the wc-1 and wc-2 genes. orthologs of wc-1 and wc-2 have hitherto been found only in filamentous fungi and not in yeast, with the exception of the basidiomycete pathogenic yeast cryptococcus. here, we report that the fission yeast schizosaccharomyces japonicus responds to blue light depending on wcs1 and wcs2, orthologs of components ...201323873862
nuclear dynamics in a fungal chimera.a fungal colony is a syncytium composed of a branched and interconnected network of cells. chimerism endows colonies with increased virulence and ability to exploit nutritionally complex substrates. moreover, chimera formation may be a driver for diversification at the species level by allowing lateral gene transfer between strains that are too distantly related to hybridize sexually. however, the processes by which genomic diversity develops and is maintained within a single colony are little u ...201323861490
optimization of cdt-1 and xyl1 expression for balanced co-production of ethanol and xylitol from cellobiose and xylose by engineered saccharomyces cerevisiae.production of ethanol and xylitol from lignocellulosic hydrolysates is an alternative to the traditional production of ethanol in utilizing biomass. however, the conversion efficiency of xylose to xylitol is restricted by glucose repression, causing a low xylitol titer. to this end, we cloned genes cdt-1 (encoding a cellodextrin transporter) and gh1-1 (encoding an intracellular β-glucosidase) from neurospora crassa and xyl1 (encoding a xylose reductase that converts xylose into xylitol) from sch ...201323844185
simultaneous saccharification and fermentation by engineered saccharomyces cerevisiae without supplementing extracellular β-glucosidase.simultaneous saccharification and fermentation (ssf) has been considered a promising and economical process for cellulosic ethanol production. further cost savings could be gained by reducing enzyme loading and engineering host strain for ethanol production. in this study, we demonstrate efficient ethanol production by ssf without supplementation of β-glucosidase using an engineered saccharomyces cerevisiae strain expressing a cellodextrin transporter and an intracellular β-glucosidase from neur ...201323835155
sulfur-regulated control of the met-2⁺ gene of neurospora crassa encoding cystathionine β-lyase.cystathionine β-lyase performs an essential role in the transsulfuration pathway by its primary reaction of forming homocysteine from cystathionine. understanding how the neurospora crassa met-2⁺ gene, which encodes cystathionine β-lyase, is regulated is important in determining the basis of the cellular control of transsulfuration. the aim of this study was to determine the nature of a potential regulatory connection of met-2⁺ to the neurospora sulfur regulatory network.201323835025
conformational stability and crystal packing: polymorphism in neurospora crassa cat-3.polymorphism is frequently observed from different crystallization conditions. in proteins, the effect on conformational variability is poorly documented, with only a few reported examples. here, three polymorphic crystal structures determined for a large-subunit catalase, cat-3 from neurospora crassa, are reported. two of them belonged to new space groups, p1 and p43212, and a third structure belonged to the same space group, p212121, as the previously deposited 2.3 å resolution structure (pdb ...201323832201
transcriptomic profiling-based mutant screen reveals three new transcription factors mediating menadione resistance in neurospora gain insight into the regulatory mechanisms of oxidative stress responses in filamentous fungi, the genome-wide transcriptional response of neurospora crassa to menadione was analysed by digital gene expression (dge) profiling, which identified 779 upregulated genes and 576 downregulated genes. knockout mutants affecting 130 highly-upregulated genes were tested for menadione sensitivity, which revealed that loss of the transcription factor siderophore regulation (sre) (a transcriptional repre ...201323809652
residual recombination in neurospora crassa spo11 deletion homozygotes occurs during meiosis.spo11 is considered responsible for initiation of meiotic recombination in higher organisms, but previous analysis using spo11 (rip) mutants suggests that the his-3 region of neurospora crassa experiences spo11-independent recombination. however, despite possessing several stop codons, it is conceivable that the mutants are not completely null. also, since lack of spo11 interferes with chromosomal pairing and proper segregation at meiosis i, spores can be partially diploid for a period after mei ...201323801409
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
evolution, selection and isolation: a genomic view of speciation in fungal plant pathogens.895 i. 895 ii. 896 iii. 898 iv. 900 v. 902 vi. 904 vii. 905 905 references 905 summary: speciation of fungal plant pathogens has been associated with host jumps, host domestication, clonal divergence, and hybridization. although we have substantial insight into the speciation histories of several important plant pathogens, we still know very little about the underlying genetics of reproductive isolation. studies in saccharomyces cerevisiae, neurospora crassa, and nonfungal model systems illustra ...201323782262
homology modeling, ligand docking and in silico mutagenesis of neurospora hsp80 (90): insight into intrinsic atpase activity.the hsp90 family of proteins is an important component of the cellular response to elevated temperatures, environmental or physiological stress and nuclear receptor signalling. the primary object of this work is the 80-kda heat shock protein, a member of the hsp90 family, from the model filamentous fungus neurospora crassa, (henceforth referred to as hsp80nc). in contrast to more extensively characterized members of the same family, (e.g. hsp82sc of saccharomyces cerevisiae) it exhibits a higher ...201323735900
transcriptional profiling of neurospora crassa δmak-2 reveals that mitogen-activated protein kinase mak-2 participates in the phosphate signaling pathway.the filamentous fungus neurospora crassa is an excellent model system for examining molecular responses to ambient signals in eukaryotic microorganisms. inorganic phosphate (pi) is an essential growth-limiting nutrient in nature and is crucial for the synthesis of nucleic acids and the flow of genetic information. the genetic and molecular mechanisms controlling the response to pi starvation in n. crassa include at least four genes (nuc-2, preg, pogv, and nuc-1), which are involved in a hierarch ...201323733042
the plasma membrane proton pump pma-1 is incorporated into distal parts of the hyphae independently of the spitzenkörper in neurospora crassa.most models for fungal growth have proposed a directional traffic of secretory vesicles to the hyphal apex, where they temporarily aggregate at the spitzenkörper before they fuse with the plasma membrane (pm). the pm h(+)-translocating atpase (pma-1) is delivered via the classical secretory pathway (endoplasmic reticulum [er] to golgi) to the cell surface, where it pumps h(+) out of the cell, generating a large electrochemical gradient that supplies energy to h(+)-coupled nutrient uptake systems ...201323729384
the trichoderma atroviride photolyase-encoding gene is transcriptionally regulated by non-canonical light response elements.the blr-1 and blr-2 proteins of trichoderma atroviride are the neurospora crassa homologs of white collar-1 and -2, two transcription factors involved in the regulation of genes by blue light. blr-1 and blr-2 are essential for photoinduction of phr-1, a photolyase-encoding gene whose promoter exhibits sequences similar to well-characterized light regulatory elements of neurospora, including the albino proximal element and the light response element (lre). however, despite the fact that this gene ...201323721733
septins are important for cell polarity, septation and asexual spore formation in neurospora crassa and show different patterns of localisation at germ tube tips.septins are gtp-binding cytoskeletal proteins that contribute to cell polarity, vesicle trafficking, cytokinesis and cell morphogenesis. here we have characterised the six septins encoded by the genome of the model filamentous fungus neurospora crassa. analysis of septin null mutants demonstrated that septins limit the sites of emergence of germ tubes and are important for septation and conidiation in n. crassa. septins constituted a range of different higher-order structures in n. crassa - ring ...201323691103
an apparent increase in meiotic silencing strength in crosses involving inbred neurospora crassa strains.meiotic silencing by unpaired dna is a presumed rnai-mediated elimination of the transcripts of any gene that is not properly paired with a homolog in meiosis. wild-isolated strains of neurospora crassa can be classified into three types based on the apparent strength of meiotic silencing of the bml (β-tubulin) and mei-3 genes in crosses of the wild strains with the tester strains ::bml(r) and ::mei-3. crosses with "or" and "sad" type wild-isolates, respectively, did or did not silence both the ...201323685173
reduced glutathione export during programmed cell death of neurospora a previous study, we demonstrated that staurosporine (sts) induces programmed cell death (pcd) in the fungus neurospora crassa and that glutathione has the capability of inhibiting both sts-induced reactive oxygen species (ros) formation and cell death. here, we further investigated the role of glutathione in sts-induced pcd in n. crassa and observed an efflux of reduced glutathione (gsh) together with a change in the cell internal redox state to a more oxidative environment. this event was a ...201323666236
novel insights into the role of neurospora crassa ndufaf2, an evolutionarily conserved mitochondrial complex i assembly factor.complex i deficiency is commonly associated with mitochondrial oxidative phosphorylation diseases. mutations in nuclear genes encoding structural subunits or assembly factors of complex i have been increasingly identified as the cause of the diseases. one such factor, ndufaf2, is a paralog of the ndufa12 structural subunit of the enzyme, but the mechanism by which it exerts its function remains unknown. herein, we demonstrate that the neurospora crassa ndufaf2 homologue, the 13.4 l protein, is a ...201323648483
the bacterial secondary metabolite 2,4-diacetylphloroglucinol impairs mitochondrial function and affects calcium homeostasis in neurospora crassa.the bacterial secondary metabolite 2,4-diacetylphloroglucinol (dapg) is of interest as an active ingredient of biological control strains of pseudomonas fluorescens and as a potential lead pharmaceutical molecule because of its capacity to inhibit growth of diverse microbial and non-microbial cells. the mechanism by which this occurs is unknown and in this study the filamentous fungus neurospora crassa was used as a model to investigate the effects of dapg on a eukaryotic cell. colony growth, co ...201323624246
the type of mutations induced by carbon-ion-beam irradiation of the filamentous fungus neurospora crassa.heavy-ion beams are known to cause great damage to cellular components and are particularly renowned for their ability to generate dna double-strand breaks (dsbs). to gain insight into the mutagenic effect of carbon-ion beams and how such damage is repaired by the cell, neurospora crassa mutants deficient in one of three components involved in the repair of dsbs, nonhomologous end-joining (nhej), homologous recombination repair (hr), and the mre11-rad50-xrs2 (mrx) complex, were irradiated with a ...201323622717
production of xylitol by recombinant microalgae.microalgae have received significant attention recently as a potential low-cost host for the production of next-generation biofuels and natural products. here we show that the chloroplast genome of the eukaryotic green microalga chlamydomonas reinhardtii can be genetically engineered to produce xylitol through the introduction of a gene encoding a xylose reductase (xr) from the fungi neurospora crassa. increased levels of heterologous protein accumulation and xylitol production were achieved by ...201323597921
the transcriptional response to nonself in the fungus podospora fungi, heterokaryon incompatibility is a nonself recognition process occurring when filaments of different isolates of the same species fuse. compatibility is controlled by so-called het loci and fusion of strains of unlike het genotype triggers a complex incompatibility reaction that leads to the death of the fusion cell. herein, we analyze the transcriptional changes during the incompatibility reaction in podospora anserina. the incompatibility response was found to be associated with a mas ...201323589521
computational discovery of transcriptional regulatory modules in fungal ribosome biogenesis genes reveals novel sequence and function patterns.genes involved in ribosome biogenesis and assembly (rba) are responsible for ribosome formation. in saccharomyces cerevisiae, their transcription is regulated by two dissimilar dna motifs. we were interested in analyzing conservation and divergence of rba transcription regulation machinery throughout fungal evolution. we have identified orthologs of s. cerevisiae rba genes in 39 species across fungal phylogeny and searched upstream regions of these gene sets for dna sequences significantly simil ...201323555806
biochemical characterization of molybdenum cofactor-free nitrate reductase from neurospora crassa.nitrate reductase (nr) is a complex molybdenum cofactor (moco)-dependent homodimeric metalloenzyme that is vitally important for autotrophic organism as it catalyzes the first and rate-limiting step of nitrate assimilation. beside moco, eukaryotic nr also binds fad and heme as additional redox active cofactors, and these are involved in electron transfer from nad(p)h to the enzyme molybdenum center where reduction of nitrate to nitrite takes place. we report the first biochemical characterizatio ...201323539622
regional control of histone h3 lysine 27 methylation in neurospora.trimethylated lysine 27 on histone h3 (h3k27me3) is present in drosophila, arabidopsis, worms, and mammals, but is absent from yeasts that have been examined. we identified and analyzed h3k27me3 in the filamentous fungus neurospora crassa and in other neurospora species. h3k27me3 covers 6.8% of the n. crassa genome, encompassing 223 domains, including 774 genes, all of which are transcriptionally silent. n. crassa h3k27me3-marked genes are less conserved than unmarked genes and only ∼35% of gene ...201323530226
trans-species activity of a nonself recognition domain.the ability to distinguish nonself from self is a fundamental characteristic of biological systems. in the filamentous fungus neurospora crassa, multiple incompatibility genes mediate nonself recognition during vegetative growth. one of these genes, un-24, encodes both nonself recognition function and the large subunit of a type i ribonucleotide reductase, an evolutionarily conserved enzyme that is essential for the conversion of ndp precursors into dndps for use in dna synthesis. previous studi ...201323517247
alternative splicing in ascomycetes.alternative splicing is a complex and regulated process, which results in mrna with different coding capacities from a single gene. extend and types of alternative splicing vary greatly among eukaryotes. in this review, i focus on alternative splicing in ascomycetes, which in general have significant lower extend of alternative splicing than mammals. yeast-like species have low numbers of introns and consequently alternative splicing is lower compared to filamentous fungi. several examples from ...201323515838
novel proteins required for meiotic silencing by unpaired dna and sirna generation in neurospora crassa.during meiosis in the filamentous fungus neurospora crassa, unpaired genes are identified and silenced by a process known as meiotic silencing by unpaired dna (msud). previous work has uncovered six proteins required for msud, all of which are also essential for meiotic progression. additionally, they all localize in the perinuclear region, suggesting that it is a center of msud activity. nevertheless, at least a subset of msud proteins must be present inside the nucleus, as unpaired dna recogni ...201323502675
identification of small rnas associated with meiotic silencing by unpaired neurospora crassa, unpaired genes are silenced by a mechanism called meiotic silencing by unpaired dna (msud). although some rna interference proteins are necessary for this process, its requirement of small rnas has yet to be formally established. here we report the characterization of small rnas targeting an unpaired region, using illumina sequencing.201323502674
coordination of k+ transporters in neurospora: trk1 is scarce and constitutive, while hak1 is abundant and highly regulated.fungi, plants, and bacteria accumulate potassium via two distinct molecular machines not directly coupled to atp hydrolysis. the first, designated trk, hkt, or ktr, has eight transmembrane helices and is folded like known potassium channels, while the second, designated hak, kt, or kup, has 12 transmembrane helices and resembles mfs class proteins. one of each type functions in the model organism neurospora crassa, where both are readily accessible for biochemical, genetic, and electrophysiologi ...201323475706
biotransformation of 14-deoxy-14-methylenetriptolide into a novel hydroxylation product by neurospora crassa.the biotransformation of 14-deoxy-14-methylenetriptolide by neurospora crassa cgmcc as 3.1604 to produce a new hydroxylation derivative was studied. the structure of this novel compound was determined using spectral data. this biotransformation using whole cells conditioned for 4 days transformed approximately 65% (mol ratio) of the substrate into the compound (5r)-5-hydroxy-14-deoxy-14-methylenetriptolide.201323465510
genetics of cordyceps and related fungi.ascomycete cordyceps sensu lato consists of hundreds of species of fungi capable of infecting different insects. species of these fungi are either valued traditional chinese medicines or used for biocontrol of insect pests. phylogenomic analysis indicated that fungal entomopathogenicity has evolved for multiple times, and the species of cordyceps were diverged from the mycoparasite or plant endophyte. relative to plant pathogens and saprophytes, cordyceps species demonstrate characteristic genom ...201323435902
direct cellobiose production from cellulose using sextuple beta-glucosidase gene deletion neurospora crassa cellobiose production from cellulose by a genetically modified fungus-neurospora crassa, was explored in this study. a library of n. crassa sextuple beta-glucosidase (bgl) gene deletion strains was constructed. various concentrations of cellobiose were detected in the culture broth of the n. crassa sextuple beta-glucosidase (bgl) gene deletion strains when grown on avicel without exogenous cellulase addition. the sextuple bgl deletion strains expressing one of the three basally transcribe ...201323410930
the stringency of start codon selection in the filamentous fungus neurospora eukaryotic cells initiation may occur from near-cognate codons that differ from aug by a single nucleotide. the stringency of start codon selection impacts the efficiency of initiation at near-cognate codons and the efficiency of initiation at aug codons in different contexts. we used a codon-optimized firefly luciferase reporter initiated with aug or each of the nine near-cognate codons in preferred context to examine the stringency of start codon selection in the model filamentous fungus ne ...201323396971
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