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activation of a trp-like channel and intracellular ca2+ dynamics during phospholipase-c-mediated cell death.the model organism neurospora crassa undergoes programmed cell death when exposed to staurosporine. here, we show that staurosporine causes defined changes in cytosolic free ca(2+) ([ca(2+)]c) dynamics and a distinct ca(2+) signature that involves ca(2+) influx from the external medium and internal ca(2+) stores. we investigated the molecular basis of this ca(2+) response by using [ca(2+)]c measurements combined with pharmacological and genetic approaches. phospholipase c was identified as a piv ...201425037570
genome-wide characterization of light-regulated genes in neurospora crassa.the filamentous fungus neurospora crassa responds to light in complex ways. to thoroughly study the transcriptional response of this organism to light, rna-seq was used to analyze capped and polyadenylated mrna prepared from mycelium grown for 24 hr in the dark and then exposed to light for 0 (control) 15, 60, 120, and 240 min. more than three-quarters of all defined protein coding genes (79%) were expressed in these cells. the increased sensitivity of rna-seq compared with previous microarray s ...201425053707
genetic control of anastomosis in podospora anserina.we developed a new microscopy procedure to study anastomoses in the model ascomycete podospora anserina and compared it with the previous method involving the formation of balanced heterokaryons. both methods showed a good correlation. heterokaryon formation was less quantifiable, but enabled to observe very rare events. microscopic analysis evidenced that anastomoses were greatly influence by growth conditions and were severely impaired in the idc mutants of the pampk1, pampk2, idc1 and panox1 ...201425064063
the evolutionary history of the genes involved in the biosynthesis of the antioxidant ergothioneine.ergothioneine (egt) is a histidine betaine derivative that exhibits antioxidant action in humans. egt is primarily synthesized by fungal species and a number of bacterial species. a five-gene cluster (egta, egtb, egtc, egtd &egte) responsible for egt production in mycobacteria smegmatis has recently been identified. the first fungal biosynthetic egt gene (ncegt-1) has also been identified in neurospora crassa. ncegt-1 contains domains similar to those found in m. smegmatis egtb and egtd. egt is ...201425068406
structural and functional organization of growing tips of neurospora crassa hyphae.data are presented on a variety of intracellular structures of the vegetative hyphae of the filamentous fungus neurospora crassa and the involvement of these structures in the tip growth of the hyphae. current ideas on the molecular and genetic mechanisms of tip growth and regulation of this process are considered. on the basis of comparison of data on behaviors of mitochondria and microtubules and data on the electrical heterogeneity of the hyphal apex, a hypothesis is proposed about a possible ...201425108323
two pentatricopeptide repeat domain proteins are required for the synthesis of respiratory complex i.in this study pentatricopeptide repeat (ppr) proteins in filamentous ascomycetes are identified and functionally characterized. ppr proteins, which have in common a degenerated 35 amino acid motif often arranged in multiple tandems, are known to be implicated in various steps of rna metabolism in mitochondria and chloroplasts. in filamentous ascomycetes we identified a common set of nine ppr proteins. for seven of these proteins, which were not yet characterized, knockout mutants of neurospora c ...201525108509
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
deciphering the uniqueness of mucoromycotina cell walls by combining biochemical and phylogenomic approaches.most fungi from the mucoromycotina lineage occur in ecosystems as saprobes, although some species are phytopathogens or may induce human mycosis. mucoromycotina represent early diverging models that are most valuable for understanding fungal evolution. here we reveal the uniqueness of the cell wall structure of the mucoromycotina rhizopus oryzae and phycomyces blakesleeanus compared with the better characterized cell wall of the ascomycete neurospora crassa. we have analysed the corresponding po ...201525143134
vib1, a link between glucose signaling and carbon catabolite repression, is essential for plant cell wall degradation by neurospora crassa.filamentous fungi that thrive on plant biomass are the major producers of hydrolytic enzymes used to decompose lignocellulose for biofuel production. although induction of cellulases is regulated at the transcriptional level, how filamentous fungi sense and signal carbon-limited conditions to coordinate cell metabolism and regulate cellulolytic enzyme production is not well characterized. by screening a transcription factor deletion set in the filamentous fungus neurospora crassa for mutants una ...201425144221
efficient detection of unpaired dna requires a member of the rad54-like family of homologous recombination proteins.meiotic silencing by unpaired dna (msud) is a process that detects unpaired regions between homologous chromosomes and silences them for the duration of sexual development. while the phenomenon of msud is well recognized, the process that detects unpaired dna is poorly understood. in this report, we provide two lines of evidence linking unpaired dna detection to a physical search for dna homology. first, we have found that a putative snf2-family protein (sad-6) is required for efficient msud in ...201425146971
light-mediated participation of the vivid-like protein of fusarium fujikuroi vvda in pigmentation and development.the phytopathogen fusarium fujikuroi is a model fungus for the production of different secondary metabolites. these include the acidic xanthophyll neurosporaxanthin, produced through the activity of the enzymes encoded by the car genes, of which carra and carb play a major role. expression of these genes is induced by light but, in contrast to other fungi, the induction is not impaired under continuous illumination in null mutants of the f. fujikuroi wc-1-like gene, wcoa. therefore, we investiga ...201425154020
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
regulated dna methylation and the circadian clock: implications in cancer.since the cloning and discovery of dna methyltransferases (dnmt), there has been a growing interest in dna methylation, its role as an epigenetic modification, how it is established and removed, along with the implications in development and disease. in recent years, it has become evident that dynamic dna methylation accompanies the circadian clock and is found at clock genes in neurospora, mice and cancer cells. the relationship among the circadian clock, cancer and dna methylation at clock gen ...201425198253
a family of starch-active polysaccharide monooxygenases.the recently discovered fungal and bacterial polysaccharide monooxygenases (pmos) are capable of oxidatively cleaving chitin, cellulose, and hemicelluloses that contain β(1→4) linkages between glucose or substituted glucose units. they are also known collectively as lytic pmos, or lpmos, and individually as aa9 (formerly gh61), aa10 (formerly cbm33), and aa11 enzymes. pmos share several conserved features, including a monocopper center coordinated by a bidentate n-terminal histidine residue and ...201425201969
efficient gene targeting in ligase iv-deficient monascus ruber m7 by perturbing the non-homologous end joining pathway.inactivating the non-homologous end joining (nhej) pathway is a well established method to increase gene replacement frequency (grf) in filamentous fungi because nhej is predominant for the repair of dna double strand breaks (dsbs), while gene targeting is based on homologous recombination (hr). dna ligase iv, a component of the nhej system, is strictly required for the nhej in saccharomyces cerevisiae and neurospora crassa. to enhance the grf in monascus ruber m7, we deleted the mrlig4 gene enc ...201425209642
reaction-diffusion systems for spatio-temporal intracellular protein networks: a beginner's guide with two examples.spatio-temporal dynamics of a variety of proteins is, among other things, regulated by post-translational modifications of these proteins. such modifications can thus influence stability and biochemical activities of the proteins, activity and stability of their upstream targets within specific signalling pathways. commonly used mathematical tools for such protein-protein (and/or protein-mrna) interactions in single cells, namely, michaelis-menten and hill kinetics, yielding a system of ordinary ...201425210594
a high-throughput gene disruption methodology for the entomopathogenic fungus metarhizium robertsii.systematic gene disruption is a direct way to interrogate a fungal genome to functionally characterize the full suite of genes involved in various biological processes. metarhizium robertsii is extraordinarily versatile, and it is a pathogen of arthropods, a saprophyte and a beneficial colonizer of rhizospheres. thus, m. robertsii can be used as a representative to simultaneously study several major lifestyles that are not shared by the "model" fungi saccharomyces cerevisiae and neurospora crass ...201425222118
neurospora crassa as a model organism to explore the interconnected network of the cell cycle and the circadian clock.budding and fission yeast pioneered uncovering molecular mechanisms of eukaryotic cell division cycles. however, they do not possess canonical circadian clock machinery that regulates physiological processes with a period of about 24h. on the other hand, neurospora crassa played a critical role in elucidating molecular mechanisms of circadian rhythms, but have not been utilized frequently for cell cycle studies. recent findings demonstrate that there exists a conserved coupling between the cell ...201425239547
a protein kinase screen of neurospora crassa mutant strains reveals that the snf1 protein kinase promotes glycogen synthase phosphorylation.glycogen functions as a carbohydrate reserve in a variety of organisms and its metabolism is highly regulated. the activities of glycogen synthase and glycogen phosphorylase, the rate-limiting enzymes of the synthesis and degradation processes, respectively, are regulated by allosteric modulation and reversible phosphorylation. to identify the protein kinases affecting glycogen metabolism in neurospora crassa, we performed a screen of 84 serine/threonine kinase knockout strains. we identified mu ...201425253091
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
characterizing the roles of cryphonectria parasitica rna-dependent rna polymerase-like genes in antiviral defense, viral recombination and transposon transcript accumulation.an inducible rna-silencing pathway, involving a single dicer protein, dcl2, and a single argonaute protein, agl2, was recently shown to serve as an effective antiviral defense response in the chestnut blight fungus cryphonectria parasitica. eukaryotic rna-dependent rna polymerases (rdrps) are frequently involved in transcriptional and posttranscriptional gene silencing and antiviral defense. we report here the identification and characterization of four rdrp genes (rdr1-4) in the c. parasitica g ...201425268858
bioinformatic and biochemical characterizations of c-s bond formation and cleavage enzymes in the fungus neurospora crassa ergothioneine biosynthetic pathway.ergothioneine is a histidine thiol derivative. its mycobacterial biosynthetic pathway has five steps (egta-e catalysis) with two novel reactions: a mononuclear nonheme iron enzyme (egtb) catalyzed oxidative c-s bond formation and a plp-mediated c-s lyase (egte) reaction. our bioinformatic and biochemical analyses indicate that the fungus neurospora crassa has a more concise ergothioneine biosynthetic pathway because its nonheme iron enzyme, egt1, makes use of cysteine instead of γ-glu-cys as the ...201425275953
characterization of the neurospora crassa cell fusion proteins, ham-6, ham-7, ham-8, ham-9, ham-10, amph-1 and whi-2.intercellular communication of vegetative cells and their subsequent cell fusion is vital for different aspects of growth, fitness, and differentiation of filamentous fungi. cell fusion between germinating spores is important for early colony establishment, while hyphal fusion in the mature colony facilitates the movement of resources and organelles throughout an established colony. approximately 50 proteins have been shown to be important for somatic cell-cell communication and fusion in the mo ...201425279949
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
role of a remote leucine residue in the catalytic function of polyol dehydrogenase.studies on the protein-metal binding sites have mainly focused on the residues immediately surrounding the reacting substrate, cofactors, and metal ions. the contribution of residues in remote layers to the highly optimized microenvironments of catalytic active sites is not well understood. to improve our understanding, the present study examined the role of remote residues on the structure and function of zinc-dependent polyol dehydrogenases. we used an integrated computational and biochemical ...201425292207
nothing special in the specialist? draft genome sequence of cryomyces antarcticus, the most extremophilic fungus from antarctica.the draft genome of the antarctic endemic fungus cryomyces antarcticus is presented. this rock inhabiting, microcolonial fungus is extremely stress tolerant and it is a model organism for exobiology and studies on stress resistance in eukaryots. since this fungus is a specialist in the most extreme environment of the earth, the analysis of its genome is of important value for the understanding of fungal genome evolution and stress adaptation. a comparison with neurospora crassa as well as with o ...201425296285
mechanism of the neurospora circadian clock, a frequency-centric view.circadian clocks are self-sustaining timekeepers found in almost all organisms on earth. the filamentous fungus neurospora crassa is a preeminent model for eukaryotic circadian clocks. investigations of the neurospora circadian clock system have led to elucidation of circadian clock regulatory mechanisms that are common to all eukaryotes. in this work, we will focus on the neurospora circadian oscillator mechanism with an emphasis on the regulation of the core clock component frequency.201525302868
gene expression differences among three neurospora species reveal genes required for sexual reproduction in neurospora crassa.many fungi form complex three-dimensional fruiting bodies, within which the meiotic machinery for sexual spore production has been considered to be largely conserved over evolutionary time. indeed, much of what we know about meiosis in plant and animal taxa has been deeply informed by studies of meiosis in saccharomyces and neurospora. nevertheless, the genetic basis of fruiting body development and its regulation in relation to meiosis in fungi is barely known, even within the best studied mult ...201425329823
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
the transcription factor ste12 mediates the regulatory role of the tmk1 map kinase in mycoparasitism and vegetative hyphal fusion in the filamentous fungus trichoderma atroviride.mycoparasitic species of the fungal genus trichoderma are potent antagonists able to combat plant pathogenic fungi by direct parasitism. an essential step in this mycoparasitic fungus-fungus interaction is the detection of the fungal host followed by activation of molecular weapons in the mycoparasite by host-derived signals. the trichoderma atroviride map kinase tmk1, a homolog of yeast fus3/kss1, plays an essential role in regulating the mycoparasitic host attack, aerial hyphae formation and c ...201425356841
cellulose surface degradation by a lytic polysaccharide monooxygenase and its effect on cellulase hydrolytic efficiency.lytic polysaccharide monooxygenase (lpmo) represents a unique principle of oxidative degradation of recalcitrant insoluble polysaccharides. used in combination with hydrolytic enzymes, lpmo appears to constitute a significant factor of the efficiency of enzymatic biomass depolymerization. lpmo activity on different cellulose substrates has been shown from the slow release of oxidized oligosaccharides into solution, but an immediate and direct demonstration of the enzyme action on the cellulose s ...201425361767
a novel cryptochrome-dependent oscillator in neurospora crassa.several lines of evidence suggest that the circadian clock is constructed of multiple molecular feedback oscillators that function to generate robust rhythms in organisms. however, while core oscillator mechanisms driving specific behaviors are well described in several model systems, the nature of other potential circadian oscillators is not understood. using genetic approaches in the fungus neurospora crassa, we uncovered an oscillator mechanism that drives rhythmic spore development in the ab ...201525361899
analysis of clock-regulated genes in neurospora reveals widespread posttranscriptional control of metabolic potential.neurospora crassa has been for decades a principal model for filamentous fungal genetics and physiology as well as for understanding the mechanism of circadian clocks. eukaryotic fungal and animal clocks comprise transcription-translation-based feedback loops that control rhythmic transcription of a substantial fraction of these transcriptomes, yielding the changes in protein abundance that mediate circadian regulation of physiology and metabolism: understanding circadian control of gene express ...201425362047
the cullin-4 complex dcdc does not require e3 ubiquitin ligase elements to control heterochromatin in neurospora crassa.the cullin-4 (cul4) complex dcdc (dim-5/-7/-9/cul4/ddb1 complex) is essential for dna methylation and heterochromatin formation in neurospora crassa. cullins form the scaffold of cullin-ring e3 ubiquitin ligases (crls) and are modified by the covalent attachment of nedd8, a ubiquitin-like protein that regulates the stability and activity of crls. we report that neddylation is not required for cul4-dependent dna methylation or heterochromatin formation but is required for the dna repair functions ...201525362134
engineering neurospora crassa for improved cellobiose and cellobionate production.we report engineering neurospora crassa to improve the yield of cellobiose and cellobionate from cellulose. a previously engineered strain of n. crassa (f5) with six of seven β-glucosidase (bgl) genes knocked out was shown to produce cellobiose and cellobionate directly from cellulose without the addition of exogenous cellulases. in this study, the f5 strain was further modified to improve the yield of cellobiose and cellobionate from cellulose by increasing cellulase production and decreasing p ...201525381238
isolation, characterization and application of a cellulose-degrading strain neurospora crassa s1 from oil palm empty fruit bunch.oil palm empty fruit bunch (efb) is a lignocellulosic waste produced in palm oil industry. efb mainly consists of cellulose, hemicellulose (mainly xylan) and lignin and has a great potential to be reused. converting efb to fermentable sugars and value-added chemicals is a much better choice than treating efb as waste.201425384340
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
the putative cellodextrin transporter-like protein clp1 is involved in cellulase induction in neurospora crassa.neurospora crassa recently has become a novel system to investigate cellulase induction. here, we discovered a novel membrane protein, cellodextrin transporter-like protein 1 (clp1; ncu05853), a putative cellodextrin transporter-like protein that is a critical component of the cellulase induction pathway in n. crassa. although clp1 protein cannot transport cellodextrin, the suppression of cellulase induction by this protein was discovered on both cellobiose and avicel. the co-disruption of the c ...201525398875
fungal communication requires the mak-2 pathway elements ste-20 and ras-2, the nrc-1 adapter ste-50 and the map kinase scaffold ham-5.intercellular communication is critical for the survival of unicellular organisms as well as for the development and function of multicellular tissues. cell-to-cell signaling is also required to develop the interconnected mycelial network characteristic of filamentous fungi and is a prerequisite for symbiotic and pathogenic host colonization achieved by molds. somatic cell-cell communication and subsequent cell fusion is governed by the mak-2 mitogen activated protein kinase (mapk) cascade in th ...201425411845
ham-5 functions as a map kinase scaffold during cell fusion in neurospora crassa.cell fusion in genetically identical neurospora crassa germlings and in hyphae is a highly regulated process involving the activation of a conserved map kinase cascade that includes nrc-1, mek-2 and mak-2. during chemotrophic growth in germlings, the map kinase cascade members localize to conidial anastomosis tube (cat) tips every ∼8 minutes, perfectly out of phase with another protein that is recruited to the tip: soft, a recently identified scaffold for the mak-1 map kinase pathway in sordaria ...201425412208
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
development and physiological characterization of cellobiose-consuming yarrowia lipolytica.yarrowia lipolytica is a promising production host for a wide range of molecules, but limited sugar consumption abilities prevent utilization of an abundant source of renewable feedstocks. in this study we created a y. lipolytica strain capable of utilizing cellobiose as a sole carbon source by using endogenous promoters to express the cellodextrin transporter cdt-1 and intracellular β-glucosidase gh1-1 from neurospora crassa. the engineered strain was also capable of simultaneous co-consumption ...201525421388
biomineralization of metal carbonates by neurospora crassa.in this research, the urease-positive fungus neurospora crassa was investigated for the biomineralization of calcium carbonate and its potential application in metal biorecovery and/or bioremediation. after 12 d incubation at 25 °c in urea and calcium-containing medium, extensive biomineralization of fungal filaments was observed. energy dispersive x-ray analysis of crystalline precipitates on the hyphae of n. crassa showed that the main elements present in the crystals were ca, c, and o. x-ray ...201425423300
the rab gtpase ypt-1 associates with golgi cisternae and spitzenkörper microvesicles in neurospora crassa.vesicle traffic involves budding, transport, tethering and fusion of vesicles with acceptor membranes. gtp-bound small rab gtpases interact with the membrane of vesicles, promoting their association with other factors before their subsequent fusion. filamentous fungi contain at their hyphal apex the spitzenkörper (spk), a multivesicular structure to which vesicles concentrate before being redirected to specific cell sites. the regulatory mechanisms ensuring the directionality of the vesicles tha ...201525425138
ascomycete fungal actins differentially support plant spatial cell and organ development.actin interacts with a wide variety of cytoplasmic and nuclear proteins to support spatial development in nearly all eukaryotes. null mutations in plant vegetative actins produce dramatically altered cell, tissue, and organ morphologies. animal cytoplasmic actins (e.g., human hsactb, hsactg1) and some ancestral protist actins fully suppress these mutant phenotypes suggesting that some animal, plant, and protist actins share functional competence for spatial development. considering that fungi ha ...201525428798
the histone h3 lysine 9 methyltransferase dim-5 modifies chromatin at frequency and represses light-activated gene expression.the transcriptional program controlling the circadian rhythm requires coordinated regulation of chromatin. characterization of the chromodomain helicase dna-binding enzyme chd1 revealed dna methylation in the promoter of the central clock gene frequency (frq) in neurospora crassa. in this report, we show that the dna methylation at frq is not only dependent on the dna methyltransferase dim-2 but also on the h3k9 methyltransferase dim-5 and hp1. histone h3 lysine 9 trimethylation (h3k9me3) occurs ...201425429045
endogenous ergothioneine is required for wild type levels of conidiogenesis and conidial survival but does not protect against 254 nm uv-induced mutagenesis or kill.ergothioneine, a histidine derivative, is concentrated in conidia of ascomycetous fungi. to investigate the function of ergothioneine, we crossed the wild type neurospora crassa (egt(+)) and an ergothioneine non-producer (egt(-), δegt-1, a knockout in ncu04343.5) and used the egt(+) and egt(-) progeny strains for phenotypic analyses. compared to the egt(+) strains, egt(-) strains had a 59% reduction in the number of conidia produced on vogel's agar. after storage of egt(+) and egt(-) conidia at ...201425446508
the chestnut blight fungus for studies on virus/host and virus/virus interactions: from a natural to a model host.the chestnut blight fungus, cryphonectria parasitica, is an important plant pathogenic ascomycete. the fungus hosts a wide range of viruses and now has been established as a model filamentous fungus for studying virus/host and virus/virus interactions. this is based on the development of methods for artificial virus introduction and elimination, host genome manipulability, available host genome sequence with annotations, host mutant strains, and molecular tools. molecular tools include sub-cellu ...201525454384
kinesin-3 in the basidiomycete ustilago maydis transports organelles along the entire microtubule array.the molecular motor kinesin-3 transports early endosomes along microtubules in filamentous fungi. it was reported that kinesin-3 from the ascomycete fungi aspergillus nidulans and neurospora crassa use a subset of post-translationally modified and more stable microtubules. here, i show that kinesin-3 from the basidiomycete ustilago maydis moves along all hyphal microtubules. this difference is likely due to variation in cell cycle control and associated organization of the microtubule array.201525459534
selection and evaluation of reference genes for expression studies with quantitative pcr in the model fungus neurospora crassa under different environmental conditions in continuous culture.neurospora crassa has served as a model organism for studying circadian pathways and more recently has gained attention in the biofuel industry due to its enhanced capacity for cellulase production. however, in order to optimize n. crassa for biotechnological applications, metabolic pathways during growth under different environmental conditions must be addressed. reverse-transcription quantitative pcr (rt-qpcr) is a technique that provides a high-throughput platform from which to measure the ex ...201425474155
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
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
[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
[lah-3, a transcriptional factor involved in osmotic regulation in neurospora crassa].[objective] in order to identify the function of lah-3 in osmotic regulation, we generated lah-3 deletion strain and analyzed its phenotype by osmostress treatment. [methods] we used homologous recombination to replace lah-3gene by hph gene and treated these cells with 4% nacl and 1 m sorbitol to analyze the phenotype. northern blot was used to detect the expressions of osmoresponsing genes. western blot was used to examine the phosphorylation level of lah-3 and os-2 and the expression of os-2. ...201425522587
the apses family proteins in fungi: characterizations, evolution and functions.the apses protein family belongs to transcriptional factors of the basic helix-loop-helix (bhlh) class, the originally described members (apses: asm1p, phd1p, sok2p, efg1p and stuap) are used to designate this group of proteins, and they have been identified as key regulators of fungal development and other biological processes. apses proteins share a highly conserved dna-binding domain (apses domain) of about 100 amino acids, whose central domain is predicted to form a typical bhlh structure. b ...201525534868
the aging biological clock in neurospora crassa.the biological clock affects aging through ras-1 (bd) and lag-1, and these two longevity genes together affect a clock phenotype and the clock oscillator in neurospora crassa. using an automated cell-counting technique for measuring conidial longevity, we show that the clock-associated genes lag-1 and ras-1 (bd) are true chronological longevity genes. for example, wild type (wt) has an estimated median life span of 24 days, while the double mutant lag-1, ras-1 (bd) has an estimated median life s ...201425535564
evolution of rna-protein interactions: non-specific binding led to rna splicing activity of fungal mitochondrial tyrosyl-trna synthetases.the neurospora crassa mitochondrial tyrosyl-trna synthetase (mttyrrs; cyt-18 protein) evolved a new function as a group i intron splicing factor by acquiring the ability to bind group i intron rnas and stabilize their catalytically active rna structure. previous studies showed: (i) cyt-18 binds group i introns by using both its n-terminal catalytic domain and flexibly attached c-terminal anticodon-binding domain (ctd); and (ii) the catalytic domain binds group i introns specifically via multiple ...201425536042
probing c-terminal interactions of the pseudomonas stutzeri cyanide-degrading cynd protein.the cyanide dihydratases from bacillus pumilus and pseudomonas stutzeri share high amino acid sequence similarity throughout except for their highly divergent c-termini. however, deletion or exchange of the c-termini had different effects upon each enzyme. here we extended previous studies and investigated how the c-terminus affects the activity and stability of three nitrilases, the cyanide dihydratases from b. pumilus (cyndpum) and p. stutzeri (cyndstut) and the cyanide hydratase from neurospo ...201525549622
neurospora crassa asm-1 complements the conidiation defect in a stua mutant of aspergillus nidulans.aspergillus nidulans stua and neurospora crassa asm-1 are orthologous apses (asm-1, phd1, sok2, efg1, stua) transcription factors conserved across a diverse group of fungi. stua and asm-1 have roles in asexual (conidiation) and sexual (ascospore formation) development in both organisms. to address the hypothesis that the last common ancestor of these diverse fungi regulated conidiation with similar genes, asm-1 was introduced into the stua1 mutant of a. nidulans. expression of asm-1 complemented ...201525550299
hypotonic stress-induced calcium signaling in saccharomyces cerevisiae involves trp-like transporters on the endoplasmic reticulum membrane.saccharomyces cerevisiae cells respond to hypotonic stress (hts) by a cytosolic calcium rise, either generated by an influx of calcium from extracellular medium, when calcium is available, or by a release from intracellular stores in scarcity of extracellular calcium. calcium release from intracellular compartments is peculiarly inhibited by external calcium in a calcineurin-independent and cch1-, but not mid1-, driven manner. hts-induced calcium release is also negatively regulated by the er pr ...201525573187
identification and characterization of lfd-2, a predicted fringe protein required for membrane integrity during cell fusion in neurospora crassa.the molecular mechanisms of membrane merger during somatic cell fusion in eukaryotic species are poorly understood. in the filamentous fungus neurospora crassa, somatic cell fusion occurs between genetically identical germinated asexual spores (germlings) and between hyphae to form the interconnected network characteristic of a filamentous fungal colony. in n. crassa, two proteins have been identified to function at the step of membrane fusion during somatic cell fusion: prm1 and lfd-1. the abse ...201525595444
dissecting the function of the different chitin synthases in vegetative growth and sexual development in neurospora crassa.chitin, one of the most important carbohydrates of the fungal cell wall, is synthesized by chitin synthases (chs). seven sequences encoding chss have been identified in the genome of neurospora crassa. previously, chs-1, -3 and -6 were found at the spitzenkörper(spk) core and developing septa. we investigated the functional importance of each chs in growth and development of n. crassa. the cellular distribution of each chs tagged with fluorescent proteins and the impact of corresponding gene del ...201525596036
prevalent and distinct spliceosomal 3'-end processing mechanisms for fungal telomerase rna.telomerase rna (ter) is an essential component of the telomerase ribonucleoprotein complex. the mechanism for ter 3'-end processing is highly divergent among different organisms. here we report a unique spliceosome-mediated ter 3'-end cleavage mechanism in neurospora crassa that is distinct from that found specifically in the fission yeast schizosaccharomyces pombe. while the s. pombe ter intron contains the canonical 5'-splice site guaugu, the n. crassa ter intron contains a non-canonical 5'-sp ...201525598218
circadian rhythms. decoupling circadian clock protein turnover from circadian period determination.the mechanistic basis of eukaryotic circadian oscillators in model systems as diverse as neurospora, drosophila, and mammalian cells is thought to be a transcription-and-translation-based negative feedback loop, wherein progressive and controlled phosphorylation of one or more negative elements ultimately elicits their own proteasome-mediated degradation, thereby releasing negative feedback and determining circadian period length. the neurospora crassa circadian negative element frequency (frq) ...201525635104
multiple cellular roles of neurospora crassa plc-1, spla2, and cpe-1 in regulation of cytosolic free calcium, carotenoid accumulation, stress responses, and acquisition of thermotolerance.phospholipase c1 (plc1), secretory phospholipase a2 (spla2) and ca(2+)/h(+) exchanger proteins regulate calcium signaling and homeostasis in eukaryotes. in this study, we investigate functions for phospholipase c1 (plc-1), spla2 (spla2) and a ca(2+)/h(+) exchanger (cpe-1) in the filamentous fungus neurospora crassa. the δplc-1, δspla2, and δcpe-1 mutants exhibited a growth defect on medium supplemented with the divalent ionophore a23187, suggesting that these genes might play a role in regulatio ...201525636422
complex formation of rna silencing proteins in the perinuclear region of neurospora crassa.in neurospora, genes not paired during meiosis are targeted by meiotic silencing by unpaired dna (msud). here, our bimolecular fluorescence complementation (bifc) study suggests that rna-directed rna polymerase, dicer, argonaute, and others form a silencing complex in the perinuclear region, with intimate interactions among the majority of them. we have also shown that sad-2 is likely the anchor for this assembly.201525644701
expanding xylose metabolism in yeast for plant cell wall conversion to biofuels.sustainable biofuel production from renewable biomass will require the efficient and complete use of all abundant sugars in the plant cell wall. using the cellulolytic fungus neurospora crassa as a model, we identified a xylodextrin transport and consumption pathway required for its growth on hemicellulose. reconstitution of this xylodextrin utilization pathway in saccharomyces cerevisiae revealed that fungal xylose reductases act as xylodextrin reductases, producing xylosyl-xylitol oligomers as ...201525647728
transient transformation of podosphaera xanthii by electroporation of conidia.powdery mildew diseases are a major phytosanitary issue causing important yield and economic losses in agronomic, horticultural and ornamental crops. powdery mildew fungi are obligate biotrophic parasites unable to grow on culture media, a fact that has significantly limited their genetic manipulation. in this work, we report a protocol based on the electroporation of fungal conidia, for the transient transformation of podosphaera fusca (synonym podosphaera xanthii), the main causal agent of cuc ...201525651833
exploring the potential of fungal arylacetonitrilases in mandelic acid synthesis.the application of arylacetonitrilases from filamentous fungi to the hydrolysis of high concentrations of (r,s)-mandelonitrile (100-500 mm) was demonstrated for the first time. escherichia coli strains expressing the corresponding genes were used as whole-cell catalysts. nitrilases from aspergillus niger, neurospora crassa, nectria haematococca, and arthroderma benhamiae (enzymes nitan, nitnc, nitnh, and nitab, respectively) exhibited different degrees of enantio- and chemoselectivity (amide for ...201525652193
synthesis of a new glycosphingolipid, neurosporaside, from neurospora crassa.the glycosphingolipid neurosporaside (α-d-glcp-(1 → 2)-β-d-galp-(1 → 6)-β-d-galp-(1 → 6)-β-d-galp-(1 →)-cer) occurs in neurospora crassa. we attempted to synthesize neurosporaside by block synthesis (route a) and linear synthesis (route b). oligosaccharide derivatives were synthesized using trimethylsilyltrifluoromethanesulfonate and n-iodosuccinimide/trifluoromethane sulfonic acid as promoters. the target tetrasaccharide could not be attained via route a, but route b showed potential: glycosidi ...201525659120
dissecting the mechanisms of the clock in neurospora.the circadian clock exists to synchronize inner physiology with the external world, allowing life to anticipate and adapt to the continual changes that occur in an organism's environment. the clock architecture is highly conserved, present in almost all major branches of life. within eukaryotes, the filamentous fungus neurospora crassa has consistently been used as an excellent model organism to uncover the basic circadian physiology and molecular biology. the neurospora model has elucidated our ...201525662450
methods to study molecular mechanisms of the neurospora circadian clock.eukaryotic circadian clocks are comprised of interlocked autoregulatory feedback loops that control gene expression at the levels of transcription and translation. the filamentous fungus neurospora crassa is an excellent model for the complex molecular network of regulatory mechanisms that are common to all eukaryotes. at the heart of the network, posttranslational regulation and functions of the core clock elements are of major interest. this chapter discusses the methods used currently to stud ...201525662455
the role of cercospora zeae-maydis homologs of rhodobacter sphaeroides 1o2-resistance genes in resistance to the photoactivated toxin cercosporin.the photosynthetic bacterium rhodobacter sphaeroides and plant pathogenic fungus cercospora nicotianae have been used as models for understanding resistance to singlet oxygen ((1)o(2)), a highly toxic reactive oxygen species. in rhodobacter and cercospora, (1)o(2) is derived, respectively, from photosynthesis and from the (1)o(2)-generating toxin cercosporin which the fungus produces to parasitize plants. we identified common genes recovered in transcriptome studies of putative (1)o(2)-resistanc ...201525670706
experimental demonstration of the benefits of somatic fusion and the consequences for allorecognition.allorecognition, the ability to distinguish "self" from "nonself" based on allelic differences at allorecognition loci, is common in all domains of life. allorecognition restricts the opportunities for social parasitism, and is therefore crucial for the evolution of cooperation. however, the maintenance of allorecognition diversity provides a paradox. if allorecognition is costly relative to cooperation, common alleles will be favored. thus, the cost of allorecognition may reduce the genetic var ...201525688421
a transcriptomic analysis of neurospora crassa using five major crop residues and the novel role of the sporulation regulator rca-1 in lignocellulase production.crop residue is an abundant, low-cost plant biomass material available worldwide for use in the microbial production of enzymes, biofuels, and valuable chemicals. however, the diverse chemical composition and complex structure of crop residues are more challenging for efficient degradation by microbes than are homogeneous polysaccharides. in this study, the transcriptional responses of neurospora crassa to various plant straws were analyzed using rna-seq, and novel beneficial factors for biomass ...201525691917
in vitro production and antifungal activity of peptide abp-dhc-cecropin a.the antimicrobial peptide abp-dhc-cecropin a is a small cationic peptide with potent activity against a wide range of bacterial species. evidence of antifungal activity has also been suggested; however, testing of this peptide has been limited due to the low expression of cecropin proteins in escherichia coli. to improve expression of this peptide in e. coli, abp-dhc-cecropin a was cloned into a psumo vector and transformed into e. coli, resulting in the production of a psumo-abp-dhc-cecropin a ...201525702854
heterologous expression of mitochondria-targeted microbial nitrilase enzymes increases cyanide tolerance in arabidopsis.anthropogenic activities have resulted in cyanide (cn) contamination of both soil and water in many areas of the globe. while plants possess a detoxification pathway that serves to degrade endogenously generated cn, this system is readily overwhelmed, limiting the use of plants in bioremediation. genetic engineering of additional cn degradation pathways in plants is one potential strategy to increase their tolerance to cn. here we show that heterologous expression of microbial nitrilase enzymes ...201525711239
functional characterization and low-resolution structure of an endoglucanase cel45a from the filamentous fungus neurospora crassa or74a: thermostable enzyme with high activity toward lichenan and β-glucan.biomass is the most abundant and short-term renewable natural resource on earth whose recalcitrance toward enzymatic degradation represents significant challenge for a number of biotechnological applications. the not so abundant but critically necessary class of gh45 endoglucanases constitutes an essential component of tailored industrial enzyme cocktails because they randomly and internally cleave cellulose molecules. moreover, gh45 glucanases are core constituents of major-brand detergent form ...201525711741
fungal genome sequencing: basic biology to biotechnology.the genome sequences provide a first glimpse into the genomic basis of the biological diversity of filamentous fungi and yeast. the genome sequence of the budding yeast, saccharomyces cerevisiae, with a small genome size, unicellular growth, and rich history of genetic and molecular analyses was a milestone of early genomics in the 1990s. the subsequent completion of fission yeast, schizosaccharomyces pombe and genetic model, neurospora crassa initiated a revolution in the genomics of the fungal ...201625721271
clerodane type diterpene as a novel antifungal agent from polyalthia longifolia var. pendula.bioactivity-guided chemical examination of methanolic extract of leaves of polyalthia longifolia var. pendula led to the isolation of the active constituent, a diterpene 1 which was identified as 16α-hydroxycleroda-3,13(14)z-dien-15,16-olide on the basis of its spectral data. among the tested strains, diterpene 1 was found to exhibit antifungal activities having mic90 values of 50.3, 100.6 and 201.2 μm against candida albicans ncim3557, cryptococcus neoformans ncim3542 (human pathogens) and neur ...201525747495
carbon and nitrogen limitation increase chitosan antifungal activity in neurospora crassa and fungal human pathogens.chitosan permeabilizes plasma membrane and kills sensitive filamentous fungi and yeast. membrane fluidity and cell energy determine chitosan sensitivity in fungi. a five-fold reduction of both glucose (main carbon (c) source) and nitrogen (n) increased 2-fold neurospora crassa sensitivity to chitosan. we linked this increase with production of intracellular reactive oxygen species (ros) and plasma membrane permeabilization. releasing n. crassa from nutrient limitation reduced chitosan antifungal ...201525749367
dawn- and dusk-phased circadian transcription rhythms coordinate anabolic and catabolic functions in neurospora.circadian clocks control rhythmic expression of a large number of genes in coordination with the 24 hour day-night cycle. the mechanisms generating circadian rhythms, their amplitude and circadian phase are dependent on a transcriptional network of immense complexity. moreover, the contribution of post-transcriptional mechanisms in generating rhythms in rna abundance is not known.201525762222
neurospora importin α is required for normal heterochromatic formation and dna methylation.heterochromatin and associated gene silencing processes play roles in development, genome defense, and chromosome function. in many species, constitutive heterochromatin is decorated with histone h3 tri-methylated at lysine 9 (h3k9me3) and cytosine methylation. in neurospora crassa, a five-protein complex, dcdc, catalyzes h3k9 methylation, which then directs dna methylation. here, we identify and characterize a gene important for dcdc function, dim-3 (defective in methylation-3), which encodes t ...201525793375
combinatorial control of light induced chromatin remodeling and gene activation in neurospora.light is an important environmental cue that affects physiology and development of neurospora crassa. the light-sensing transcription factor (tf) wcc, which consists of the gata-family tfs wc1 and wc2, is required for light-dependent transcription. sub1, another gata-family tf, is not a photoreceptor but has also been implicated in light-inducible gene expression. to assess regulation and organization of the network of light-inducible genes, we analyzed the roles of wcc and sub1 in light-induced ...201525822411
draft genome sequence of neurospora crassa strain fgsc 73.we report the elucidation of the complete genome of the neurospora crassa (shear and dodge) strain fgsc 73, a mat-a, trp-3 mutant strain. the genome sequence around the idiotypic mating type locus represents the only publicly available sequence for a mat-a strain. 40.42 megabases are assembled into 358 scaffolds carrying 11,978 gene models.201525838471
functional analysis of two l-arabinose transporters from filamentous fungi reveals promising characteristics for improved pentose utilization in saccharomyces cerevisiae.limited uptake is one of the bottlenecks for l-arabinose fermentation from lignocellulosic hydrolysates in engineered saccharomyces cerevisiae. this study characterized two novel l-arabinose transporters, lat-1 from neurospora crassa and mtlat-1 from myceliophthora thermophila. although the two proteins share high identity (about 83%), they display different substrate specificities. sugar transport assays using the s. cerevisiae strain eby.vw4000 indicated that lat-1 accepts a broad substrate sp ...201525841015
production of cellobionate from cellulose using an engineered neurospora crassa strain with laccase and redox mediator addition.we report a novel production process for cellobionic acid from cellulose using an engineered fungal strain with the exogenous addition of laccase and a redox mediator. a previously engineered strain of neurospora crassa (f5∆ace-1∆cre-1∆ndvb) was shown to produce cellobionate directly from cellulose without the addition of exogenous cellulases. specifically, n. crassa produces cellulases, which hydrolyze cellulose to cellobiose, and cellobiose dehydrogenase (cdh), which oxidizes cellobiose to cel ...201525849253
reconstruction of the carnitine biosynthesis pathway from neurospora crassa in the yeast saccharomyces cerevisiae.industrial synthesis of l-carnitine is currently performed by whole-cell biotransformation of industrial waste products, mostly d-carnitine and cronobetaine, through specific bacterial species. no comparable system has been established using eukaryotic microorganisms, even though there is a significant and growing international demand for either the pure compound or carnitine-enriched consumables. in eukaryotes, including the fungus neurospora crassa, l-carnitine is biosynthesized through a four ...201525851717
parallel structural evolution of mitochondrial ribosomes and oxphos complexes.the five macromolecular complexes that jointly mediate oxidative phosphorylation (oxphos) in mitochondria consist of many more subunits than those of bacteria, yet, it remains unclear by which evolutionary mechanism(s) these novel subunits were recruited. even less well understood is the structural evolution of mitochondrial ribosomes (mitoribosomes): while it was long thought that their exceptionally high protein content would physically compensate for their uniquely low amount of ribosomal rna ...201525861818
membrane transporters in self resistance of cercospora nicotianae to the photoactivated toxin cercosporin.the goal of this work is to characterize membrane transporter genes in cercospora fungi required for autoresistance to the photoactivated, active-oxygen-generating toxin cercosporin they produce for infection of host plants. previous studies implicated a role for diverse membrane transporters in cercosporin resistance. in this study, transporters identified in a subtractive cdna library between a cercospora nicotianae wild type and a cercosporin-sensitive mutant were characterized, including two ...201525862648
mathematical modeling and validation of glucose compensation of the neurospora circadian clock.autonomous circadian oscillations arise from transcriptional-translational feedback loops of core clock components. the period of a circadian oscillator is relatively insensitive to changes in nutrients (e.g., glucose), which is referred to as "nutrient compensation". recently, a transcription repressor, csp-1, was identified as a component of the circadian system in neurospora crassa. the transcription of csp-1 is under the circadian regulation. intriguingly, csp-1 represses the circadian trans ...201525863073
mutagenesis of trichoderma reesei endoglucanase i: impact of expression host on activity and stability at elevated temperatures.trichoderma reesei is a key cellulase source for economically saccharifying cellulosic biomass for the production of biofuels. lignocellulose hydrolysis at temperatures above the optimum temperature of t. reesei cellulases (~50°c) could provide many significant advantages, including reduced viscosity at high-solids loadings, lower risk of microbial contamination during saccharification, greater compatibility with high-temperature biomass pretreatment, and faster rates of hydrolysis. these potent ...201525879765
inferring bona fide transfrags in rna-seq derived-transcriptome assemblies of non-model organisms.de novo transcriptome assembly of short transcribed fragments (transfrags) produced from sequencing-by-synthesis technologies often results in redundant datasets with differing levels of unassembled, partially assembled or mis-assembled transcripts. post-assembly processing intended to reduce redundancy typically involves reassembly or clustering of assembled sequences. however, these approaches are mostly based on common word heuristics and often create clusters of biologically unrelated sequen ...201525880035
genome-wide analysis of the endoplasmic reticulum stress response during lignocellulase production in neurospora crassa.lignocellulolytic fungal cells suffer endoplasmic reticulum (er) stress during lignocellulase synthesis; however, an understanding of this integrated process on a genome-wide scale remains poor. here, we undertook a systematic investigation of this process in neurospora crassa (n. crassa) using transcriptomic analysis coupled with genetic screens.201525883682
regulation of glycogen metabolism by the cre-1, rco-1 and rcm-1 proteins in neurospora crassa. the role of cre-1 as the central transcriptional regulator.the transcription factor crea/mig1/cre-1 is a repressor protein that regulates the use of alternative carbon sources via a mechanism known as carbon catabolite repression (ccr). in saccharomyces cerevisiae, mig1 recruits the complex ssn6-tup1, the neurospora crassa rcm-1 and rco-1 orthologous proteins, respectively, to bind to promoters of glucose-repressible genes. we have been studying the regulation of glycogen metabolism in n. crassa and the identification of the rco-1 corepressor as a regul ...201525889113
isobutanol production from cellobionic acid in escherichia coli.liquid fuels needed for the global transportation industry can be produced from sugars derived from plant-based lignocellulosics. lignocellulosics contain a range of sugars, only some of which (such as cellulose) have been shown to be utilizable by microorganisms capable of producing biofuels. cellobionic acid makes up a small but significant portion of lignocellulosic degradation products, and had not previously been investigated as an utilizable substrate. however, aldonic acids such as cellob ...201525889729
neurospora heterokaryons with complementary duplications and deficiencies in their constituent nuclei provide an approach to identify nucleus-limited genes.introgression is the transfer of genes or genomic regions from one species into another via hybridization and back-crosses. we have introgressed four translocations (eb4, ibj5, uk14-1, and b362i) from neurospora crassa into n. tetrasperma. this enabled us to construct two general types of heterokaryons with mat-a and mat-a nuclei of different genotypes: one type is [t + n] (with one translocation nucleus and one normal sequence nucleus), and the other is [dp + df] (with one nucleus carrying a du ...201525897010
enzymatic characterization of recombinant nitrate reductase expressed and purified from neurospora crassa.we established an expression and purification procedure for recombinant protein production in neurospora crassa (n. crassa). this strep-tag® based system was successfully used for purifying recombinant n. crassa nitrate reductase (nr), whose enzymatic activity was compared to recombinant n. crassa nr purified from escherichia coli. the purity of the two different nr preparations was similar but nr purified from n. crassa showed a significantly higher nitrate turnover rate. two phosphorylation si ...201525914160
a lipid-managing program maintains a stout spitzenkörper.the spitzenkörper (spk) is an accumulation of vesicles interleaved with actin microfilaments present at the cytosolic side of the apical plasma membrane (pm) of hyphal tips of many species of filamentous fungi. the physiological role of the spk has captivated fungal biologists over the years, but only very recently this 'organelle' is starting to be understood in the molecular terminology used for cell biological models. one aspect that has received little attention is the role of cellular membr ...201525921726
improvement of omega-3 docosahexaenoic acid production by marine dinoflagellate crypthecodinium cohnii using rapeseed meal hydrolysate and waste molasses as feedstock.rapeseed meal and waste molasses are two important agro-industrial by-products which are produced in large quantities. in this study, solid state fermentation and fungal autolysis were performed to produce rapeseed meal hydrolysate (rmh) using fungal strains of aspergillus oryzae, penicillium oxalicum and neurospora crassa. the hydrolysate was used as fermentation feedstock for heterotrophic growth of microalga crypthecodinium cohnii that produce docosahexaenoic acid (dha). the addition of waste ...201525942565
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