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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 anserina.in 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 dna.in 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 mutants.direct 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 crassa.in 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
global analyses of ceratocystis cacaofunesta mitochondria: from genome to proteome.the ascomycete fungus ceratocystis cacaofunesta is the causal agent of wilt disease in cacao, which results in significant economic losses in the affected producing areas. despite the economic importance of the ceratocystis complex of species, no genomic data are available for any of its members. given that mitochondria play important roles in fungal virulence and the susceptibility/resistance of fungi to fungicides, we performed the first functional analysis of this organelle in ceratocystis us ...201323394930
both osrecq1 and osrdr1 are required for the production of small rna in response to dna-damage in rice.small rna-mediated gene silencing pathways play important roles in the regulation of development, genome stability and various stress responses in many eukaryotes. recently, a new type of small interfering rnas (qirnas) approximately 20-21 nucleotides long in neurospora crassa have been shown to mediate gene silencing in the dna damage response (ddr) pathway. however, the mechanism for rna silencing in the ddr pathway is largely unknown in plants. here, we report that a class of small rnas (qirn ...201323383126
eukaryotic tpp riboswitch regulation of alternative splicing involving long-distance base pairing.thiamin pyrophosphate (tpp) riboswitches are found in organisms from all three domains of life. examples in bacteria commonly repress gene expression by terminating transcription or by blocking ribosome binding, whereas most eukaryotic tpp riboswitches are predicted to regulate gene expression by modulating rna splicing. given the widespread distribution of eukaryotic tpp riboswitches and the diversity of their locations in precursor messenger rnas (pre-mrnas), we sought to examine the mechanism ...201323376932
glucoamylase is a potential allergen in products expressed in neurospora crassa. 201323352537
transcription of the major neurospora crassa microrna-like small rnas relies on rna polymerase iii.most plant and animal micrornas (mirnas) are transcribed by rna polymerase ii. we previously discovered mirna-like small rnas (milrnas) in the filamentous fungus neurospora crassa and uncovered at least four different pathways for milrna production. to understand the evolutionary origin of milrnas, we determined the roles of polymerases ii and iii (pol ii and pol iii) in milrna transcription. our results show that pol iii is responsible for the transcription of the major milrnas produced in this ...201323349642
using a model filamentous fungus to unravel mechanisms of lignocellulose deconstruction.filamentous fungi are the main source of enzymes used to degrade lignocellulose to fermentable sugars for the production of biofuels. while the most commonly used organism for the production of cellulases in an industrial setting is trichoderma reesei (hypocrea jecorina), recent work in the model filamentous fungus neurospora crassa has shown that the variety of molecular, genetic and biochemical techniques developed for this organism can expedite analyses of the complexities involved in the uti ...201323339486
nonself recognition through intermolecular disulfide bond formation of ribonucleotide reductase in neurospora.type i ribonucleotide reductases (rnrs) are conserved across diverse taxa and are essential for the conversion of rna into dna precursors. in neurospora crassa, the large subunit of rnr (un-24) is unusual in that it also has a nonself recognition function, whereby coexpression of oak ridge (or) and panama (pa) alleles of un-24 in the same cell leads to growth inhibition and cell death. we show that coexpressing these incompatible alleles of un-24 in n. crassa results in a high molecular weight u ...201323335337
biotechnological production of ethanol from renewable resources by neurospora crassa: an alternative to conventional yeast fermentations?microbial production of ethanol might be a potential route to replace oil and chemical feedstocks. bioethanol is by far the most common biofuel in use worldwide. lignocellulosic biomass is the most promising renewable resource for fuel bioethanol production. bioconversion of lignocellulosics to ethanol consists of four major unit operations: pretreatment, hydrolysis, fermentation, and product separation/distillation. conventional bioethanol processes for lignocellulosics apply commercial fungal ...201323318834
reactive oxygen species can modulate circadian phase and period in neurospora crassa.reactive oxygen species (ros) may serve as signals coupling metabolism to other cell functions. in addition to being by-products of normal metabolism, they are generated at elevated levels under environmental stress situations. we analyzed how reactive oxygen species affect the circadian clock in the model organism neurospora crassa. in light/dark cycles, an increase in the levels of reactive oxygen species advanced the phase of both the conidiation rhythm and the expression of the clock gene fr ...201323277144
deletion and expression analysis of beta-(1,3)-glucanosyltransferase genes in neurospora crassa.gpi(glycosylphosphatidylinositol)-anchored beta-(1,3)-glucanosyltransferases play an active role in cell wall biosynthesis in fungi. neurospora crassa has 5 putative beta-(1,3)-glucanosyltransferase genes, namely, gel-1, gel-2, gel-3, gel-4, and gel-5, in its genome. among them, the gel-3 gene is constitutively expressed at the highest level in growing hyphae, whereas gel-1 is expressed at the lowest level. the gel-3 deletion mutant displayed slow growth, while other gel gene disruptants exhibit ...201323274249
[organization of mitochondria in the growing hyphae of neurospora crassa].in vivo fluorescent labeling of mitochondria in neurospora crassa showed the concentration of filamentous mitochondria within 30 μm of apex in growing hyphae. these mitochondrial assemblies propagated forward with the elongation of hyphae, split and segregated as the growing tip bifurcated and formed de novo when new branches formed farther away from the apex. the efficiency of the mitochondria concentration in the apical 30 μm zone is related to the growth rate and identical in hyphae cultivate ...201325509139
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
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
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
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
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
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 crassa.in 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
phylogenetics and gene structure dynamics of polygalacturonase genes in aspergillus and neurospora crassa.polygalacturonase (pg) gene is a typical gene family present in eukaryotes. forty-nine pgs were mined from the genomes of neurospora crassa and five aspergillus species. the pgs were classified into 3 clades such as clade 1 for rhamno-pgs, clade 2 for exo-pgs and clade 3 for exo- and endo-pgs, which were further grouped into 13 sub-clades based on the polypeptide sequence similarity. in gene structure analysis, a total of 124 introns were present in 44 genes and five genes lacked introns to give ...201325288950
microbial transformations of isophorone by alternaria alternata and neurospora crassa.isophorone (3,5,5-trimethyl-2-cyclohexen-1-one), a monoterpene, and the structurally related 1,8-cineole and camphor, have demonstrated a protective effect against cancer, biological activity against a variety of microorganisms, and anti-oxidant properties. the derivatization of isophorone is, therefore, an important field of xenobiochemistry, pharmacology and toxicology. the aim of this study was to obtain derivatives of isophorone through microbial biotransformation and evaluate the biotransfo ...201323472460
phosphorylation of calcineurin at a novel serine-proline rich region orchestrates hyphal growth and virulence in aspergillus fumigatus.the fungus aspergillus fumigatus is a leading infectious killer in immunocompromised patients. calcineurin, a calmodulin (cam)-dependent protein phosphatase comprised of calcineurin a (cnaa) and calcineurin b (cnab) subunits, localizes at the hyphal tips and septa to direct a. fumigatus invasion and virulence. here we identified a novel serine-proline rich region (sprr) located between two conserved cnaa domains, the cnab-binding helix and the cam-binding domain, that is evolutionarily conserved ...201323990785
fungal siderophore biosynthesis is partially localized in peroxisomes.siderophores play a central role in iron metabolism and virulence of most fungi. both aspergillus fumigatus and aspergillus nidulans excrete the siderophore triacetylfusarinine c (tafc) for iron acquisition. in a. fumigatus, green fluorescence protein-tagging revealed peroxisomal localization of the tafc biosynthetic enzymes sidi (mevalonyl-coa ligase), sidh (mevalonyl-coa hydratase) and sidf (anhydromevalonyl-coa transferase), while elimination of the peroxisomal targeting signal (pts) impaired ...201323617799
fungal cell wall organization and biosynthesis.the composition and organization of the cell walls from saccharomyces cerevisiae, candida albicans, aspergillus fumigatus, schizosaccharomyces pombe, neurospora crassa, and cryptococcus neoformans are compared and contrasted. these cell walls contain chitin, chitosan, β-1,3-glucan, β-1,6-glucan, mixed β-1,3-/β-1,4-glucan, α-1,3-glucan, melanin, and glycoproteins as major constituents. a comparison of these cell walls shows that there is a great deal of variability in fungal cell wall composition ...201323419716
two functional motifs define the interaction, internalization and toxicity of the cell-penetrating antifungal peptide paf26 on fungal cells.the synthetic, cell penetrating hexapeptide paf26 (rkkwfw) is antifungal at low micromolar concentrations and has been proposed as a model for cationic, cell-penetrating antifungal peptides. its short amino acid sequence facilitates the analysis of its structure-activity relationships using the fungal models neurospora crassa and saccharomyces cerevisiae, and human and plant pathogens aspergillus fumigatus and penicillium digitatum, respectively. previously, paf26 at low fungicidal concentration ...201323349973
the oxygenase cao-1 of neurospora crassa is a resveratrol cleavage enzyme.the genome of the ascomycete neurospora crassa encodes cao-1 and cao-2, two members of the carotenoid cleavage oxygenase family that target double bonds in different substrates. previous studies demonstrated the role of cao-2 in cleaving the c40 carotene torulene, a key step in the synthesis of the c35 apocarotenoid pigment neurosporaxanthin. in this work, we investigated the activity of cao-1, assuming that it may provide retinal, the chromophore of the nop-1 rhodopsin, by cleaving β-carotene. ...201323893079
molecular tools for functional genomics in filamentous fungi: recent advances and new strategies.advances in genetic transformation techniques have made important contributions to molecular genetics. various molecular tools and strategies have been developed for functional genomic analysis of filamentous fungi since the first dna transformation was successfully achieved in neurospora crassa in 1973. increasing amounts of genomic data regarding filamentous fungi are continuously reported and large-scale functional studies have become common in a wide range of fungal species. in this review, ...201323988676
the l-cysteine desulfurase nfs1 is localized in the cytosol where it provides the sulfur for molybdenum cofactor biosynthesis in humans.in humans, the l-cysteine desulfurase nfs1 plays a crucial role in the mitochondrial iron-sulfur cluster biosynthesis and in the thiomodification of mitochondrial and cytosolic trnas. we have previously demonstrated that purified nfs1 is able to transfer sulfur to the c-terminal domain of mocs3, a cytosolic protein involved in molybdenum cofactor biosynthesis and trna thiolation. however, no direct evidence existed so far for the interaction of nfs1 and mocs3 in the cytosol of human cells. here, ...201323593335
the uve1 endonuclease is regulated by the white collar complex to protect cryptococcus neoformans from uv damage.the pathogenic fungus cryptococcus neoformans uses the bwc1-bwc2 photoreceptor complex to regulate mating in response to light, virulence and ultraviolet radiation tolerance. how the complex controls these functions is unclear. here, we identify and characterize a gene in cryptococcus, uve1, whose mutation leads to a uv hypersensitive phenotype. the homologous gene in fission yeast schizosaccharomyces pombe encodes an apurinic/apyrimidinic endonuclease acting in the uvde-dependent excision repai ...201324039606
physiological role of acyl coenzyme a synthetase homologs in lipid metabolism in neurospora crassa.acyl coenzyme a (coa) synthetase (acs) enzymes catalyze the activation of free fatty acids (fas) to coa esters by a two-step thioesterification reaction. activated fas participate in a variety of anabolic and catabolic lipid metabolic pathways, including de novo complex lipid biosynthesis, fa β-oxidation, and lipid membrane remodeling. analysis of the genome sequence of the filamentous fungus neurospora crassa identified seven putative fatty acss (acs-1 through acs-7). acs-3 was found to be the ...201323873861
sterol c-22 desaturase erg5 mediates the sensitivity to antifungal azoles in neurospora crassa and fusarium verticillioides.antifungal azoles inhibit ergosterol biosynthesis by interfering with lanosterol 14α-demethylase. in this study, seven upregulated and four downregulated ergosterol biosynthesis genes in response to ketoconazole treatment were identified in neurospora crassa. azole sensitivity test of knockout mutants for six ketoconazole-upregulated genes in ergosterol biosynthesis revealed that deletion of only sterol c-22 desaturase erg5 altered sensitivity to azoles: the erg5 mutant was hypersensitive to azo ...201323755044
a comparative study of hydrolysis and transglycosylation activities of fungal β-glucosidases.β-glucosidases (bgs) from aspergillus fumigatus, aspergillus niger, aspergillus oryzae, magnaporthe grisea, neurospora crassa, and penicillium brasilianum were purified to homogeneity, and investigated for their (simultaneous) hydrolytic and transglycosylation activity in samples with high concentrations of either cellobiose or glucose. the rate of the hydrolytic process (which converts one cellobiose to two glucose molecules) shows a maximum around 10-15 mm cellobiose and decreases with further ...201322311644
assessing the effects of light on differentiation and virulence of the plant pathogen botrytis cinerea: characterization of the white collar complex.organisms are exposed to a tough environment, where acute daily challenges, like light, can strongly affect several aspects of an individual's physiology, including pathogenesis. while several fungal models have been widely employed to understand the physiological and molecular events associated with light perception, various other agricultural-relevant fungi still remain, in terms of their responsiveness to light, in the dark. the fungus botrytis cinerea is an aggressive pathogen able to cause ...201324391918
proteomic alterations induced by ionic liquids in aspergillus nidulans and neurospora crassa.this study constitutes the first attempt to understand at the proteomic level the fungal response to ionic liquid stress. ascomycota are able to grow in media supplemented with high concentrations of an ionic liquid, which, in turn, lead to major alterations in the fungal metabolic footprint. herein, we analysed the differential accumulation of mycelial proteins in aspergillus nidulans and neurospora crassa after their exposure to two of the most commonly used ionic liquids: 1-ethyl-3-methylimid ...201324120530
analysis of a conserved cellulase transcriptional regulator reveals inducer-independent production of cellulolytic enzymes in neurospora crassa.cellulose is recalcitrant to deconstruction to glucose for use in fermentation strategies for biofuels and chemicals derived from lignocellulose. in neurospora crassa, the transcriptional regulator, clr-2, is required for cellulolytic gene expression and cellulose deconstruction. to assess conservation and divergence of cellulase gene regulation between fungi from different ecological niches, we compared clr-2 function with its ortholog (clrb) in the distantly related species, aspergillus nidula ...201323766336
neurospora crassa nkin2, a kinesin-3 motor, transports early endosomes and is required for polarized growth.biological motors are molecular nanomachines, which convert chemical energy into mechanical forces. the combination of mechanoenzymes with structural components, such as the cytoskeleton, enables eukaryotic cells to overcome entropy, generate molecular gradients, and establish polarity. hyphae of filamentous fungi are among the most polarized cells, and polarity defects are most obvious. here, we studied the role of the kinesin-3 motor, nkin2, in neurospora crassa. we found that nkin2 localizes ...201323687116
conf and conj contribute to conidia germination and stress response in the filamentous fungus aspergillus nidulans.light induces various responses in fungi including formation of asexual and sexual reproductive structures. the formation of conidia in the filamentous fungus aspergillus nidulans is regulated by red and blue light receptors. expression of conidia associated con genes, which are widely spread in the fungal kingdom, increases upon exposure to light. we have characterized the light-inducible conf and conj genes of a. nidulans which are homologs of con-6 and con-10 of neurospora crassa. con genes a ...201323644150
eukaryotic resistance to fluoride toxicity mediated by a widespread family of fluoride export proteins.fluorine is an abundant element and is toxic to organisms from bacteria to humans, but the mechanisms by which eukaryotes resist fluoride toxicity are unknown. the escherichia coli gene crcb was recently shown to be regulated by a fluoride-responsive riboswitch, implicating it in fluoride response. there are >8,000 crcb homologs across all domains of life, indicating that it has an important role in biology. here we demonstrate that eukaryotic homologs [renamed fex (fluoride exporter)] function ...201324173035
effect of heterologous xylose transporter expression in candida tropicalis on xylitol production rate.xylose utilization is inhibited by glucose uptake in xylose-assimilating yeasts, including candida tropicalis, resulting in limitation of xylose uptake during the fermentation of glucose/xylose mixtures. in this study, a heterologous xylose transporter gene (at5g17010) from arabidopsis thaliana was selected because of its high affinity for xylose and was codon-optimized for functional expression in c. tropicalis. the codon-optimized gene was placed under the control of the gapdh promoter and was ...201323411871
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
aspergillus oryzae aoso is a novel component of stress granules upon heat stress in filamentous fungi.stress granules are a type of cytoplasmic messenger ribonucleoprotein (mrnp) granule formed in response to the inhibition of translation initiation, which typically occurs when cells are exposed to stress. stress granules are conserved in eukaryotes; however, in filamentous fungi, including aspergillus oryzae, stress granules have not yet been defined. for this reason, here we investigated the formation and localization of stress granules in a. oryzae cells exposed to various stresses using an e ...201323991062
a comparative study of nitrilases identified by genome mining.escherichia coli strains expressing different nitrilases transformed nitriles or kcn. six nitrilases (from aspergillus niger (2), a. oryzae, neurospora crassa, arthroderma benhamiae, and nectria haematococca) were arylacetonitrilases, two enzymes (from a. niger and penicillium chrysogenum) were cyanide hydratases and the others (from p. chrysogenum, p. marneffei, gibberella moniliformis, meyerozyma guilliermondi, rhodococcus rhodochrous, and r. ruber) preferred (hetero)aromatic nitriles as subst ...201323475593
[on the biosynthesis of inorganic polyphosphates in neurospora crassa]. 20134300424
[the uptake of orthophosphate by adenine-deficient mutant of neurospora crassa 286-10 hsa]. 20135599898
mutagenicity of methylhydroxylamines in neurospora crassa. 20136054956
[on the localization of inorganic polyphosphates and nucleotides in neurospora crassa mycelium]. 20135999969
fungal type iii polyketide synthases.this article covers the literature on fungal type iii polyketide synthases (pkss) published from 2005 to 2014. since the first discovery of fungal type iii pks genes in aspergillus oryzae, reported in 2005, putative genes for type iii pkss have been discovered in fungal genomes. compared with type i pkss, type iii pkss are much less abundant in fungi. however, type iii pkss could have some critical roles in fungi. this article summarizes the studies on fungal type iii pks functional analysis, in ...201425182423
a large nonconserved region of the tethering protein leashin is involved in regulating the position, movement, and function of woronin bodies in aspergillus oryzae.the woronin body is a pezizomycotina-specific organelle that is typically tethered to the septum, but upon hyphal wounding, it plugs the septal pore to prevent excessive cytoplasmic loss. leashin (lah) is a large woronin body tethering protein that contains highly conserved n- and c-terminal regions and a long (∼2,500-amino-acid) nonconserved middle region. as the involvement of the nonconserved region in woronin body function has not been investigated, here, we functionally characterized indivi ...201424813188
efficient formation of heterokaryotic sclerotia in the filamentous fungus aspergillus oryzae.heterokaryon formation by hyphal fusion occurs during a sexual/parasexual cycle in filamentous fungi, and therefore, it is biotechnologically important for crossbreeding. in the industrial filamentous fungus aspergillus oryzae, a parasexual cycle has been reported, and it was recently suggested that sexual reproduction should be possible. however, as a. oryzae enters into hyphal fusion with a much lower frequency than neurospora crassa, the process of heterokaryon formation has not been extensiv ...201424201891
sensitivity of neurospora crassa to a marine-derived aspergillus tubingensis anhydride exhibiting antifungal activity that is mediated by the mas1 protein.the fungus aspergillus tubingensis (strain oy907) was isolated from the mediterranean marine sponge ircinia variabilis. extracellular extracts produced by this strain were found to inhibit the growth of several fungi. among the secreted extract components, a novel anhydride metabolite, tubingenoic anhydride a (1) as well as the known 2-carboxymethyl-3-hexylmaleic acid anhydride, asperic acid, and campyrone a and c were purified and their structure elucidated. compound 1 and 2-carboxymethyl-3-hex ...201425257783
the neurospora crassa cps-1 polysaccharide synthase functions in cell wall biosynthesis.the neurospora crassa cps-1 gene encodes a polysaccharide synthase with homology to the cryptococcus neoformans hyaluronic acid synthase cps1p. homologs of the cps-1 gene are found in the genomes of many fungi. loss of cps-1 results in a cell wall defect that affects all stages of the n. crassa life cycle, including vegetative growth, protoperithecia (female mating structure) development, and conidia (asexual spore) development. the cell wall of cps-1 deletion mutants is sensitive to cell wall p ...201424953997
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
nitrogen regulation of fungal secondary metabolism in fungi.fungi occupy diverse environments where they are constantly challenged by stressors such as extreme ph, temperature, uv exposure, and nutrient deprivation. nitrogen is an essential requirement for growth, and the ability to metabolize a wide variety of nitrogen sources enables fungi to colonize different environmental niches and survive nutrient limitations. favored nitrogen sources, particularly ammonium and glutamine, are used preferentially, while the expression of genes required for the use ...201425506342
genetic characterization of the neurospora crassa molybdenum cofactor biosynthesis.molybdenum (mo) is a trace element that is essential for important cellular processes. to gain biological activity, mo must be complexed in the molybdenum cofactor (moco), a pterin derivative of low molecular weight. moco synthesis is a multi-step pathway that involves a variable number of genes in eukaryotes, which are assigned to four steps of eukaryotic moco biosynthesis. moco biosynthesis mutants lack any moco-dependent enzymatic activities, including assimilation of nitrate (plants and fung ...201424569084
the transcription factor bcltf1 regulates virulence and light responses in the necrotrophic plant pathogen botrytis cinerea.botrytis cinerea is the causal agent of gray mold diseases in a range of dicotyledonous plant species. the fungus can reproduce asexually by forming macroconidia for dispersal and sclerotia for survival; the latter also participate in sexual reproduction by bearing the apothecia after fertilization by microconidia. light induces the differentiation of conidia and apothecia, while sclerotia are exclusively formed in the absence of light. the relevance of light for virulence of the fungus is not o ...201424415947
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
the aspergillus nidulans zn(ii)2cys6 transcription factor an5673/rhar mediates l-rhamnose utilization and the production of α-l-rhamnosidases.various plant-derived substrates contain l-rhamnose that can be assimilated by many fungi and its liberation is catalyzed by α-l-rhamnosidases. initial data obtained in our laboratory focussing on two aspergillus nidulans α-l-rhamnosidase genes (rhaa and rhae) showed α-l-rhamnosidase production to be tightly controlled at the level of transcription by the carbon source available. whilst induction is effected by l-rhamnose, unlike many other glycosyl hydrolase genes repression by glucose and othe ...201425416526
neurospora crassa female development requires the pacc and other signal transduction pathways, transcription factors, chromatin remodeling, cell-to-cell fusion, and autophagy.using a screening protocol we have identified 68 genes that are required for female development in the filamentous fungus neurospora crassa. we find that we can divide these genes into five general groups: 1) genes encoding components of the pacc signal transduction pathway, 2) other signal transduction pathway genes, including genes from the three n. crassa map kinase pathways, 3) transcriptional factor genes, 4) autophagy genes, and 5) other miscellaneous genes. complementation and rip studies ...201425333968
bem46 shows eisosomal localization and association with tryptophan-derived auxin pathway in neurospora crassa.bem46 proteins are evolutionarily conserved, but their functions remain elusive. we reported previously that the bem46 protein in neurospora crassa is targeted to the endoplasmic reticulum (er) and is essential for ascospore germination. in the present study, we established a bem46 knockout strain of n. crassa. this δbem46 mutant exhibited a level of ascospore germination lower than that of the wild type but much higher than those of the previously characterized bem46-overexpressing and rna inte ...201424928924
interdependence of the actin and the microtubule cytoskeleton during fungal growth.cell polarization is a theme in biology conserved from bacteria to man. one of the most extremely polarized cells in nature is the hyphae of filamentous fungi. a continuous flow of secretion vesicles from the hyphal cell body to the tip is essential for cell wall and membrane extension. microtubules (mts) and actin, along with their corresponding motor proteins, are involved in the secretion process. therefore, the arrangement of the cytoskeleton is a crucial step to establish and maintain polar ...201424879477
exploring microrna-like small rnas in the filamentous fungus fusarium oxysporum.rna silencing such as quelling and meiotic silencing by unpaired dna (msud) and several other classes of special small rnas have been discovered in filamentous fungi recently. more than four different mechanisms of microrna-like rnas (milrnas) production have been illustrated in the model fungus neurospora crassa including a dicer-independent pathway. to date, very little work focusing on small rnas in fungi has been reported and no universal or particular characteristic of milrnas were defined ...201425141304
neurospora crassa: looking back and looking forward at a model microbe.investigation of the red bread mold that contaminated french bakeries nearly two centuries ago has led to a wealth of discoveries that have impacted our understanding of genetic, biochemical, and molecular mechanisms in microbes, from mendelian genetics and the gene-enzyme relationship to circadian rhythm and plant cell wall degradation. early neurospora research focused on elucidating mechanisms of genetic recombination and gene action and later progressed to addressing complex biological quest ...201425480699
long-read, whole-genome shotgun sequence data for five model organisms.single molecule, real-time (smrt) sequencing from pacific biosciences is increasingly used in many areas of biological research including de novo genome assembly, structural-variant identification, haplotype phasing, mrna isoform discovery, and base-modification analyses. high-quality, public datasets of smrt sequences can spur development of analytic tools that can accommodate unique characteristics of smrt data (long read lengths, lack of gc or amplification bias, and a random error profile le ...201425977796
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
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
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
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
spin-labeled mitochondrial lipids in neurospora crassa. 20144301148
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
molecular evolution and functional characterisation of an ancient phenylalanine ammonia-lyase gene (nnpal1) from nelumbo nucifera: novel insight into the evolution of the pal family in angiosperms.phenylalanine ammonia-lyase (pal; e.c.4.3.1.5) is a key enzyme of the phenylpropanoid pathway in plant development, and it catalyses the deamination of phenylalanine to trans-cinnamic acid, leading to the production of secondary metabolites. this enzyme has been identified in many organisms, ranging from prokaryotes to higher plants. because nelumbo nucifera is a basal dicot rich in many secondary metabolites, it is a suitable candidate for research on the phenylpropanoid pathway.201424884360
role of the alternaria alternata blue-light receptor lrea (white-collar 1) in spore formation and secondary metabolism.alternaria alternata is a filamentous fungus that causes considerable loss of crops of economically important feed and food worldwide. it produces more than 60 different secondary metabolites, among which alternariol (aoh) and altertoxin (atx) are the most important mycotoxins. we found that mycotoxin production and spore formation are regulated by light in opposite ways. whereas spore formation was largely decreased under light conditions, the production of aoh was stimulated 2- to 3-fold. atx ...201424532063
potential conservation of circadian clock proteins in the phylum nematoda as revealed by bioinformatic searches.although several circadian rhythms have been described in c. elegans, its molecular clock remains elusive. in this work we employed a novel bioinformatic approach, applying probabilistic methodologies, to search for circadian clock proteins of several of the best studied circadian model organisms of different taxa (mus musculus, drosophila melanogaster, neurospora crassa, arabidopsis thaliana and synechoccocus elongatus) in the proteomes of c. elegans and other members of the phylum nematoda. wi ...201425396739
cytochrome b in neurospora crassa mitochondria. site of translation of the heme protein. 20144273944
use of liquid nitrogen and high ionic strength for the isolation of functional polyribosomes from neurospora crassa. 20144271372
properties of two nuclease genes in neurospora crassa.two genes, nuc-1 and nuc-2, of neurospora crassa which were responsible for the nucleic acid digestion, were located on linkage group 1 and 2, respectively. a temperature-sensitive mutant (b1ts-2) was obtained from a nuc-2 mutant. nuclease mutants showed a reduced activity of nuclease n(3) which was found to be a complex consisting of nuclease n(3) (') and inhibitor molecule. nuclease n(3), nuclease n(3) (') and inhibitor were partially purified and estimated to have the approximate molecular we ...20144260181
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