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the neurospora crassa os mapk pathway-activated transcription factor asl-1 contributes to circadian rhythms in pathway responsive clock-controlled genes.the os-pathway mitogen-activated protein kinase (mapk) cascade of neurospora crassa is responsible for adaptation to osmotic stress. activation of the mapk, os-2, leads to the transcriptional induction of many genes involved in the osmotic stress response. we previously demonstrated that there is a circadian rhythm in the phosphorylation of os-2 under constant non-stress inducing conditions. additionally, several osmotic stress-induced genes are known to be regulated by the circadian clock. ther ...201222240319
The Neurospora crassa mutant Nc?Egt-1 identifies an ergothioneine biosynthetic gene and demonstrates that ergothioneine enhances conidial survival and protects against peroxide toxicity during conidial germination.Ergothioneine (EGT) is a histidine derivative with sulfur on the imidazole ring and a trimethylated amine; it is postulated to have an antioxidant function. Although EGT apparently is only produced by fungi and some prokaryotes, it is acquired by animals and plants from the environment, and is concentrated in animal tissues in cells with an EGT transporter. Monobromobimane derivatives of EGT allowed conclusive identification of EGT by LC/MS and the quantification of EGT in Colletotrichum gramini ...201122209968
AreA controls nitrogen source utilisation during both growth programs of the dimorphic fungus Penicillium marneffei.The opportunistic pathogen Penicillium marneffei displays a temperature-dependent dimorphic switching program with saprophytic hyphal growth at 25 °C and yeast growth at 37 °C. The areA gene of P. marneffei has been isolated and found to be required for the utilisation of nonpreferred nitrogen sources during both growth programs of P. marneffei, albeit to differing degrees. Based on this functional characterisation and high degree of sequence conservation with other fungal GATA factors, P. marne ...201222208609
a crossover hotspot near his-3 in neurospora crassa is a preferential recombination termination site.during analysis of 148 unselected neurospora crassa octads, an above average rate of crossing over was detected within a 360-base region near the 3' end of his-3, suggesting a hotspot for crossing over about 1.8 kb away from the recombination initiation site within cog. homozygous deletion of the 360-base region increases exchanges in his-3 and on the far side of his-3 from cog, with the heterozygote showing an intermediate increase. we conclude that recombination events initiated at cog termina ...201222203161
oxidative cleavage of cellulose by fungal copper-dependent polysaccharide monooxygenases.fungal-derived, copper-dependent polysaccharide monooxygenases (pmos), formerly known as gh61 proteins, have recently been shown to catalyze the o(2)-dependent oxidative cleavage of recalcitrant polysaccharides. different pmos isolated from neurospora crassa were found to generate oxidized cellodextrins modified at the reducing or nonreducing ends upon incubation with cellulose and cellobiose dehydrogenase. here we show that the nonreducing end product formed by an n. crassa pmo is a 4-ketoaldos ...201222188218
cross-talk between the cellular redox state and the circadian system in neurospora.the circadian system is composed of a number of feedback loops, and multiple feedback loops in the form of oscillators help to maintain stable rhythms. the filamentous fungus neurospora crassa exhibits a circadian rhythm during asexual spore formation (conidiation banding) and has a major feedback loop that includes the frequency (frq)/white collar (wc) -1 and -2 oscillator (fwo). a mutation in superoxide dismutase (sod)-1, an antioxidant gene, causes a robust and stable circadian rhythm compare ...201122164247
myosin concentration underlies cell size-dependent scalability of actomyosin ring constriction.in eukaryotes, cytokinesis is accomplished by an actomyosin-based contractile ring. although in caenorhabditis elegans embryos larger cells divide at a faster rate than smaller cells, it remains unknown whether a similar mode of scalability operates in other cells. we investigated cytokinesis in the filamentous fungus neurospora crassa, which exhibits a wide range of hyphal circumferences. we found that n. crassa cells divide using an actomyosin ring and larger rings constricted faster than smal ...201122123864
a factor in a wild isolated neurospora crassa strain enables a chromosome segment duplication to suppress repeat-induced point mutation.repeat-induced point mutation (rip) is a sexual stage-specific mutational process of neurospora crassa and other fungi that alters duplicated dna sequences. previous studies from our laboratory showed that chromosome segment duplications (dps) longer than (approx.) 300 kbp can dominantly suppress rip, presumably by titration of the rip machinery, and that although dps less than 200 kbp did not individually suppress rip, they could do so in homozygous and multiply heterozygous crosses, provided t ...201122116279
meiotic silencing in the homothallic fungus gibberella zeae.the homothallic ascomycete fungus gibberella zeae is an important pathogen on major cereal crops. the objective of this study was to determine whether meiotic silencing occurs in g. zeae. cytological studies demonstrated that gfp and rfp-fusion proteins were not detected during meiosis, both in heterozygous outcrosses and homozygous selfings. the deletion of rsp-1, a homologue used for studies on meiotic silencing of neurospora crassa, triggered abnormal ascospores from selfing, but outcrosses b ...201122115448
biochemical characterisation of a nadph-dependent carbonyl reductase from neurospora crassa reducing α- and β-keto esters.a gene encoding an nadph-dependent carbonyl reductase from neurospora crassa (nccr) was cloned and heterologously expressed in escherichia coli. the enzyme (nccr) was purified and biochemically characterised. nccr exhibited a restricted substrate spectrum towards various ketones, and the highest activity (468u/mg) was observed with dihydroxyacetone. however, nccr proved to be very selective in the reduction of different α- and β-keto esters. several compounds were converted to the corresponding ...201122113019
Expression of aspartic protease from Neurospora crassa in industrial ethanol-producing yeast and its application in ethanol production.In order to further improve the utilization rate of raw materials and increase ethanol productivity, the gene Asp, encoding aspartic protease in Neurospora crassa was cloned and expressed in industrial ethanol-producing yeast. To promote secretion of the acid protease, the gene was fused to signal sequence of the yeast a-factor gene and constitutively expressed under transcriptional control of the Saccharomyces cerevisiae PGK1 promoter. The resultant recombinant enzyme was characterized with res ...201122112824
disruption of alternative nad(p)h dehydrogenases leads to decreased mitochondrial ros in neurospora crassa.mitochondria are a main providers of high levels of energy, but also a major source of reactive oxygen species (ros) during normal oxidative metabolism. the involvement of neurospora crassa alternative nad(p)h dehydrogenases in mitochondrial ros production was evaluated. the growth responses of a series of respiratory mutants to several stress conditions revealed that disrupting alternative dehydrogenases leads to an increased tolerance to the redox cycler paraquat, with a mutant devoid of the e ...201222100504
glutamine metabolism during aerial mycelium growth of neurospora crassa.during vegetative growth, glutamine is accumulated in the mycelium of neurospora crassa. this high pool of glutamine seems to be required for aerial mycelium growth. enzymes responsible for the synthesis and catabolism of glutamine were measured before and during the partial transformation of a mycelial mat into aerial mycelium. in the transforming mycelial mat,considerable activities of the biosynthetic nadp-glutamate dehydrogenase and glutamine synthetase (predominantly β polypeptide) and also ...198422096812
glutamine requirement for aerial mycelium growth in neurospora crassa.five amino acids are accumulated during vegetative growth of neurospora crassa, particularly.during the prestationary growth phase. alanine, glutamine, glutamate, arginine and ornithine.comprised over 80% of the total amino acid pool in the mycelium. amino acid pools of different amino acid auxotrophs were followed during the partial transformation of a mycelial mat into an aerial mycelium. the mycelial mat under starvation and in direct contact with air rapidly formed aerial mycelium, which pro ...198422096811
diverse interactions mediate asymmetric incompatibility by the het-6 supergene complex in neurospora crassa.heterokaryon incompatibility (hi) in filamentous fungi is a form of nonself recognition that operates during the vegetative phase of the life cycle. one hi gene complex in neurospora crassa, the het-6 locus, comprises two incompatibility genes, het-6 and un-24, each having two allelic variants, oak ridge (or) and panama (pa). the un-24 gene also encodes the large subunit of ribonucleotide reductase while het-6 appears to be a member of a repetitive gene family with no other known function aside ...201222094057
direct transcriptional control of a p38 mapk pathway by the circadian clock in neurospora crassa.mapk signal transduction pathways are important regulators of stress responses, cellular growth, and differentiation. in neurospora, the circadian clock controls rhythms in phosphorylation of the p38-like mapk (os-2); however, the mechanism for this regulation is not known. we show that the wcc, a transcription factor and clock component, binds to the os-4 mapkkk promoter in response to light and rhythmically in constant darkness, peaking in the subjective morning. deletion of the wcc binding si ...201122087254
functional characterization and cellular dynamics of the cdc-42 - rac - cdc-24 module in neurospora crassa.rho-type gtpases are key regulators that control eukaryotic cell polarity, but their role in fungal morphogenesis is only beginning to emerge. in this study, we investigate the role of the cdc-42 - rac - cdc-24 module in neurospora crassa. rac and cdc-42 deletion mutants are viable, but generate highly compact colonies with severe morphological defects. double mutants carrying conditional and loss of function alleles of rac and cdc-42 are lethal, indicating that both gtpases share at least one c ...201122087253
illuminating the early signaling pathway of a fungal light-oxygen-voltage photoreceptor.circadian clocks are molecular timekeepers encountered in a wide variety of organisms, which allow to adapt the cell's metabolism and behavior to the daily and seasonal periods. their function is regulated by light-sensing proteins, among which vivid, a light-oxygen-voltage (lov) sensitive domain of the fungus neurospora crassa, constitutes one of the most prominent examples. although the major photochemical and structural changes during the photocycle of this photosensor have been elucidated th ...201222081493
Catalytically active filaments - pyruvate decarboxylase from Neurospora crassa. pH-controlled oligomer structure and catalytic function.Pyruvate decarboxylase is a key enzyme in organisms whose energy metabolism is based on alcoholic fermentation. The enzyme catalyses the nonoxidative decarboxylation of 2-oxo acids in the presence of the cofactors thiamine diphosphate and magnesium ions. Pyruvate decarboxylase species from yeasts and plant seeds studied to date are allosterically activated by their substrate pyruvate. However, detailed kinetic studies on the enzyme from Neurospora crassa demonstrate for the first time the lack o ...201122077835
fine-scale mapping in neurospora crassa by using genome-wide knockout strains.fine-scale genetic mapping is often hindered by the lack of adequate markers surrounding the locus of interest. in the filamentous ascomycete neurospora crassa, the genome has been sequenced and an effort has been made to generate genome-wide deletion strains for the entire gene set. accordingly, the hygromycin-resistant marker in each deletion strain can be used as a mapping locus in a classical three-point cross, along with the mapping target and a standard marker. we have demonstrated the fea ...201622067301
the neurospora crassa dcc-1 protein, a putative histidine kinase, is required for normal sexual and asexual development and carotenogenesis.two-component signaling pathways based on phosphoryl group transfer between histidine kinase and response regulator proteins regulate environmental responses in bacteria, archaea, plants, slime molds, and fungi. here we characterize a mutant form of dcc-1, a putative histidine kinase encoded by the ncu00939 gene of the filamentous fungus neurospora crassa. we show that this protein participates in the regulation of processes such as conidiation, perithecial development, and, to a certain degree, ...201122058142
Meiotic silencing by unpaired DNA is expressed more strongly in the early than the late perithecia of crosses involving most wild-isolated Neurospora crassa strains and in self-crosses of N. tetrasperma.Meiotic silencing by unpaired DNA is a presumed RNAi-mediated elimination of the transcripts of any gene that is not properly paired with a homolog in meiosis. Eighty wild-isolated strains of Neurospora crassa were classified into three types based on the apparent strength of meiotic silencing of the bml (ß-tubulin) and mei-3 genes in crosses with the ::Bml(r) and ::mei-3 tester strains. "OR" and "Sad" type wild-isolates, respectively, did or did not silence both the genes, whereas the "Esm" typ ...201122056520
neurospora crassa light signal transduction is affected by ros.in the ascomycete fungus neurospora crassa blue-violet light controls the expression of genes responsible for differentiation of reproductive structures, synthesis of secondary metabolites, and the circadian oscillator activity. a major photoreceptor in neurospora cells is wcc, a heterodimeric complex formed by the pas-domain-containing polypeptides wc-1 and wc-2, the products of genes white collar-1 and white collar-2. the photosignal transduction is started by photochemical activity of an exci ...201222046507
cellobiose dehydrogenase and a copper-dependent polysaccharide monooxygenase potentiate cellulose degradation by neurospora crassa.the high cost of enzymes for saccharification of lignocellulosic biomass is a major barrier to the production of second generation biofuels. using a combination of genetic and biochemical techniques, we report that filamentous fungi use oxidative enzymes to cleave glycosidic bonds in cellulose. deletion of cdh-1, the gene encoding the major cellobiose dehydrogenase of neurospora crassa, reduced cellulase activity substantially, and addition of purified cellobiose dehydrogenases from m. thermophi ...201122004347
detection of antifungal compounds in polygonum ferrugineum wedd. extracts by bioassay-guided fractionation. some evidences of their mode of action.polygonum ferrugineum wedd. (polygonaceae) is used to heal infected wounds and as antiseptic, antibiotic or antifungal in the traditional argentinean medicine. the present investigation was carried out to evaluate the antifungal properties of different extracts of aerial parts of polygonum ferrugineum, in order to give support to its ethnopharmacological use and to isolate the compounds responsible for the antifungal properties. the most active compounds were tested for their capacity of produci ...201122001591
modulation of fungal sensitivity to staurosporine by targeting proteins identified by transcriptional profiling.an analysis of the time-dependent genetic response to the death-inducer staurosporine was performed in neurospora crassa by transcriptional profiling. staurosporine induced two major genes encoding an abc transporter and a protein with similarity to regulatory subunits of potassium channels. the transcriptional response is dependent on the activity of a novel transcription factor. deletion mutants in differentially expressed genes displayed altered sensitivity to staurosporine, underscoring sign ...201122001288
Involvement of a helix-loop-helix transcription factor CHC-1 in CO(2)-mediated conidiation suppression in Neurospora crassa.The morphological switch from vegetative growth to conidiation in filamentous fungi is highly regulated, but the understanding of the regulatory mechanisms is limited. In this study, by screening a set of knock-out mutants corresponding to 103 transcription factor encoding genes in Neurospora crassa, a mutant was found to produce abundant conidia in race tubes in which conidiation in the wild-type strain was suppressed. The corresponding gene NCU00749 encodes a protein containing a helix-loop-he ...201122001287
Identification of the CRE-1 cellulolytic regulon in Neurospora crassa.In filamentous ascomycete fungi, the utilization of alternate carbon sources is influenced by the zinc finger transcription factor CreA/CRE-1, which encodes a carbon catabolite repressor protein homologous to Mig1 from Saccharomyces cerevisiae. In Neurospora crassa, deletion of cre-1 results in increased secretion of amylase and ß-galactosidase.201121980519
the neurospora crassa tob complex: analysis of the topology and function of tob38 and tob37.the tob or sam complex is responsible for assembling several proteins into the mitochondrial outer membrane, including all β-barrel proteins. we have identified several forms of the complex in neurospora crassa. one form contains tob55, tob38, and tob37; another contains these three subunits plus the mdm10 protein; while additional complexes contain only tob55. as previously shown for tob55, both tob37 and tob38 are essential for viability of the organism. mitochondria deficient in tob37 or tob3 ...201121980517
evolution of multicopper oxidase genes in coprophilous and non-coprophilous members of the order sordariales.multicopper oxidases (mco) catalyze the biological oxidation of various aromatic substrates and have been identified in plants, insects, bacteria, and wood rotting fungi. in nature, they are involved in biodegradation of biopolymers such as lignin and humic compounds, but have also been tested for various industrial applications. in fungi, mcos have been shown to play important roles during their life cycles, such as in fruiting body formation, pigment formation and pathogenicity. coprophilous f ...201121966247
global analysis of serine-threonine protein kinase genes in neurospora crassa.serine/threonine (s/t) protein kinases are crucial components of diverse signaling pathways in eukaryotes, including the model filamentous fungus neurospora crassa. in order to assess the importance of s/t kinases to neurospora biology, we embarked on a global analysis of 86 s/t kinase genes in neurospora. we were able to isolate viable mutants for 77 of the 86 kinase genes. of these, 57% exhibited at least one growth or developmental phenotype, with a relatively large fraction (40%) possessing ...201121965514
nuclear and genome dynamics in multinucleate ascomycete fungi.genetic variation between individuals is essential to evolution and adaptation. however, intra-organismic genetic variation also shapes the life histories of many organisms, including filamentous fungi. a single fungal syncytium can harbor thousands or millions of mobile and potentially genotypically different nuclei, each having the capacity to regenerate a new organism. because the dispersal of asexual or sexual spores propagates individual nuclei in many of these species, selection acting at ...201121959169
mitochondrial dysfunction resulting from the absence of mitochondrial porin in neurospora crassa.porin, the voltage-dependent anion-selective channel (vdac) in the mitochondrial outer membrane, contributes to metabolism and apoptosis. vdac function was investigated in neurospora, an obligate aerobe with a single porin. porinless strains are viable, with cold-sensitive growth, cytochrome deficiencies and overexpression of alternative oxidase. itraq labeling of mitochondria from a porinless strain and its progenitor revealed a small group of proteins with altered expression levels in the muta ...201221946565
array comparative genomic hybridizations: assessing the ability to recapture evolutionary relationships using an in silico approach.comparative genomic hybridization (cgh) with dna microarrays has many biological applications including surveys of copy number changes in tumorogenesis, species detection and identification, and functional genomics studies among related organisms. array cgh has also been used to infer phylogenetic relatedness among species or strains. although the use of the entire genome can be seen as a considerable advantage for use in phylogenetic analysis, few such studies have questioned the reliability of ...201121936922
Xylitol production is increased by expression of codon-optimized Neurospora crassa xylose reductase gene in Candida tropicalis.Xylose reductase (XR) is the first enzyme in D: -xylose metabolism, catalyzing the reduction of D: -xylose to xylitol. Formation of XR in the yeast Candida tropicalis is significantly repressed in cells grown on medium that contains glucose as carbon and energy source, because of the repressive effect of glucose. This is one reason why glucose is not a suitable co-substrate for cell growth in industrial xylitol production. XR from the ascomycete Neurospora crassa (NcXR) has high catalytic effici ...201121922311
Purification and characterization of heterologously expressed nitrilases from filamentous fungi.Nitrilases from Aspergillus niger CBS 513.88, A. niger K10, Gibberella moniliformis, Neurospora crassa OR74A, and Penicillium marneffei ATCC 18224 were expressed in Escherichia coli BL21-Gold (DE3) after IPTG induction. N. crassa nitrilase exhibited the highest yield of 69,000 U?L(-1) culture. Co-expression of chaperones (GroEL/ES in G. moniliformis and P. marneffei; GroEL/ES and trigger factor in N. crassa and A. niger CBS 513.88) enhanced the enzyme solubility. Specific activities of strains e ...201121892598
Structural elements of the mitochondrial preprotein-conducting channel Tom40 dissolved by bioinformatics and mass spectrometry.Most mitochondrial proteins are imported into mitochondria from the cytosolic compartment. Proteins destined for the outer or inner membrane, the inter-membrane space, or the matrix are recognized and translocated by the TOM machinery containing the specialized protein import channel Tom40. The latter is a protein with ß-barrel shape, which is suggested to have evolved from a porin-type protein. To obtain structural insights in the absence of a crystal structure the membrane topology of Tom40 fr ...201121888892
the azoarcus group i intron ribozyme misfolds and is accelerated for refolding by atp-dependent rna chaperone proteins.structured rnas traverse complex energy landscapes that include valleys representing misfolded intermediates. in neurospora crassa and saccharomyces cerevisiae, efficient splicing of mitochondrial group i and ii introns requires the dead box proteins cyt-19 and mss116p, respectively, which promote folding transitions and function as general rna chaperones. to test the generality of rna misfolding and the activities of dead box proteins in vitro, here we measure native folding of a small group i ...201121878649
genetic architecture of a reinforced, postmating, reproductive isolation barrier between neurospora species indicates evolution via natural selection.a role for natural selection in reinforcing premating barriers is recognized, but selection for reinforcement of postmating barriers remains controversial. organisms lacking evolvable premating barriers can theoretically reinforce postmating isolation, but only under restrictive conditions: parental investment in hybrid progeny must inhibit subsequent reproduction, and selected postmating barriers must restore parents' capacity to reproduce successfully. we show that reinforced postmating isolat ...201121876674
a widespread class of reverse transcriptase-related cellular genes.reverse transcriptases (rts) polymerize dna on rna templates. they fall into several structurally related but distinct classes and form an assemblage of rt-like enzymes that, in addition to rts, also includes certain viral rna-dependent rna polymerases (rdrp) synthesizing rna on rna templates. it is generally believed that most rt-like enzymes originate from retrotransposons or viruses and have no specific function in the host cell, with telomerases being the only notable exception. here we repo ...201121876125
genetic dissection of parylation in the filamentous fungus neurospora crassa.parylation is a posttranslational protein modification carried out by par polymerases (parps). these enzymes function as adp-ribose transferases that add polymers of adp-ribose (par) to target proteins. parp proteins have critical functions impacting the aspects of normal human health, such as aging, as well as disease development, particularly cancer. recently, the powerful antitumor parp inhibitor olaparib was shown to be effective in blocking the progression of brca1/2-associated tumors, prom ...201121870276
structure of a light-activated lov protein dimer that regulates transcription.light, oxygen, or voltage (lov) protein domains are present in many signaling proteins in bacteria, archaea, protists, plants, and fungi. the lov protein vivid (vvd) of the filamentous fungus neurospora crassa enables the organism to adapt to constant or increasing amounts of light and facilitates proper entrainment of circadian rhythms. here, we determined the crystal structure of the fully light-adapted vvd dimer and reveal the mechanism by which light-driven conformational change alters the o ...201121868352
expression and functional characterisation of tnc, a high-affinity nickel transporter from neurospora crassa.our previous in silico studies identified a high-affinity nickel transporter, tnc, from the metal transportome of neurospora crassa. a knockout mutant of the tnc gene in n. crassa failed to transport nickel, showed phenotypic growth defects and diminished urease activity under physiological levels of nickel. transport assays conducted in wild type and knockout mutant strains showed that tnc transports nickel with high affinity but exhibits selectivity for other transition metal ions like cobalt. ...201121840412
regulation of stomatal tropism and infection by light in cercospora zeae-maydis: evidence for coordinated host/pathogen responses to photoperiod?cercospora zeae-maydis causes gray leaf spot of maize, which has become one of the most widespread and destructive diseases of maize in the world. c. zeae-maydis infects leaves through stomata, which is predicated on the ability of the pathogen to perceive stomata and reorient growth accordingly. in this study, the discovery that light was required for c. zeae-maydis to perceive stomata and infect leaves led to the identification of crp1, a gene encoding a putative blue-light photoreceptor homol ...201121829344
circadian timekeeping in neurospora crassa and synechococcus elongatus.at first, the saprophytic eukaryote neurospora crassa and the photosynthetic prokaryote synechococcus elongatus may seem to have little in common. however, in both organisms a circadian clock organizes cellular biochemistry, and each organism lends itself to classical and molecular genetic investigations that have revealed a detailed picture of the molecular basis of circadian rhythmicity. in the present chapter, an overview of the molecular clockwork in each organism will be described, highligh ...201121819383
analysis of al-2 mutations in neurospora.the orange pigmentation of the fungus neurospora crassa is due to the accumulation of the xanthophyll neurosporaxanthin and precursor carotenoids. two key reactions in the synthesis of these pigments, the formation of phytoene from geranylgeranyl pyrophosphate and the introduction of +¦ cycles in desaturated carotenoid products, are catalyzed by two domains of a bifunctional protein, encoded by the gene al-2. we have determined the sequence of nine al-2 mutant alleles and analyzed the carotenoid ...201121818281
[the different effects of carbon dioxide on the toxicity of silver ions for prokaryotic and eukaryotic microorganisms].the effect of carbon dioxide on survivability of bacteria escherichia coli and the germination ability ofconidia of the fungus neurospora crassa in the presence of silver nitrate was studied. it was shown that carbon dioxide increased the toxic effect of silver ions on prokaryotic cells of e. coli but did not change the survivability of spores of the eukaryote n. crassa.201121790000
a genome-wide screen for neurospora crassa transcription factors regulating glycogen metabolism.transcription factors play a key role in transcription regulation as they recognize and directly bind to defined sites in promoter regions of target genes, and thus modulate differential expression. the overall process is extremely dynamic, as they have to move through the nucleus and transiently bind to chromatin in order to regulate gene transcription. to identify transcription factors that affect glycogen accumulation in neurospora crassa, we performed a systematic screen of a deletion strain ...201121768394
exploring the processes of dna repair and homologous integration in neurospora.this review offers a personal perspective on historical developments related to our current understanding of dna repair, recombination, and homologous integration in neurospora crassa. previous reviews have summarized and analyzed the characteristics of neurospora dna repair mutants. the early history is reviewed again here as a prelude to a discussion of the molecular cloning, annotation, gene disruption and reverse genetics of neurospora dna repair genes. the classical studies and molecular an ...201121757027
ric8 is a guanine-nucleotide exchange factor for g{alpha} subunits that regulates growth and development in neurospora crassa.heterotrimeric (aß?) g proteins are crucial components of eukaryotic signal transduction pathways. g protein coupled receptors (gpcrs) act as guanine nucleotide exchange factors (gefs) for ga subunits. recently, facilitated gdp/gtp exchange by non-gpcr gefs, such as ric8, has emerged as an important mechanism for ga regulation in animals. ric8 is present in animals and filamentous fungi, such as the model eukaryote neurospora crassa, but is absent from the genomes of baker's yeast and plants. in ...201121750256
the gene card encodes the aldehyde dehydrogenase responsible for neurosporaxanthin biosynthesis in fusarium fujikuroi.neurosporaxanthin (ß-apo-4'-carotenoic acid) biosynthesis has been studied in detail in the fungus fusarium fujikuroi. the genes and enzymes for this biosynthetic pathway are known until the last enzymatic step, the oxidation of the aldehyde group of its precursor, ß-apo-4'-carotenal. based on sequence homology to neurospora crassa ylo-1, which mediates the formation of apo-4'-lycopenoic acid from the corresponding aldehyde substrate, we cloned the card gene of f. fujikuroi and investigated the ...201121749649
solution structures of dead-box rna chaperones reveal conformational changes and nucleic acid tethering by a basic tail.the mitochondrial dead-box proteins mss116p of saccharomyces cerevisiae and cyt-19 of neurospora crassa are atp-dependent helicases that function as general rna chaperones. the helicase core of each protein precedes a c-terminal extension and a basic tail, whose structural role is unclear. here we used small-angle x-ray scattering to obtain solution structures of the full-length proteins and a series of deletion mutants. we find that the two core domains have a preferred relative orientation in ...201121746911
quantitative proteomic approach for cellulose degradation by neurospora crassa.conversion of plant biomass to soluble sugars is the primary bottleneck associated with production of economically viable cellulosic fuels and chemicals. to better understand the biochemical route that filamentous fungi use to degrade plant biomass, we have taken a quantitative proteomics approach to characterizing the secretome of neurospora crassa during growth on microcrystalline cellulose. thirteen proteins were quantified in the n. crassa secretome using a combination of absolute quantifica ...201121744778
a new mutation affecting frq-less rhythms in the circadian system of neurospora crassa.we are using the fungus neurospora crassa as a model organism to study the circadian system of eukaryotes. although the frq/wcc feedback loop is said to be central to the circadian system in neurospora, rhythms can still be seen under many conditions in frq-less (frq knockout) strains. to try to identify components of the frq-less oscillator (flo), we carried out a mutagenesis screen in a frq-less strain and selected colonies with altered conidiation (spore-formation) rhythms. a mutation we name ...201121731506
neurospora crassa homologue of neuronal calcium sensor-1 has a role in growth, calcium stress tolerance, and ultraviolet survival.ncu04379 gene encodes a conserved ca(2+) and/or calmodulin binding protein that possesses a consensus signal for n-terminal myristoylation and four ef-hands, characteristics of neuronal calcium sensor-1proteins. the ncu04379.2 knockout mutant shows slow growth rate, increased sensitivity to calcium and ultraviolet (uv) irradiation, and a wild-type fragment carrying ncu04379 gene complements the mutant. therefore, ncu04379 gene has a role in growth, calcium stress tolerance, and uv survival. cros ...201121728141
functional refolding and characterization of two tom40 isoforms from human mitochondria.tom40 proteins represent an essential class of molecules which facilitate translocation of unfolded proteins from the cytosol into the mitochondrial intermembrane space. they are part of a high-molecular mass complex that forms the protein-conducting channel in outer mitochondrial membranes. this study concerns the recombinant expression, purification and folding of amino-terminally truncated variants of the two human tom40 isoforms for structural biology experiments. both cd and ftir secondary ...201121717124
the circadian clock of neurospora crassa.circadian clocks organize our inner physiology with respect to the external world, providing life with the ability to anticipate and thereby better prepare for major fluctuations in its environment. circadian systems are widely represented in nearly all major branches of life, except archaebacteria, and within the eukaryotes, the filamentous fungus neurospora crassa has served for nearly half a century as a durable model organism for uncovering the basic circadian physiology and molecular biolog ...201221707668
cofermentation of cellobiose and galactose by an engineered saccharomyces cerevisiae strain.we demonstrate improved ethanol yield and productivity through cofermentation of cellobiose and galactose by an engineered saccharomyces cerevisiae strain expressing genes coding for cellodextrin transporter (cdt-1) and intracellular +¦-glucosidase (gh1-1) from neurospora crassa. simultaneous fermentation of cellobiose and galactose can be applied to producing biofuels from hydrolysates of marine plant biomass.201121705527
systems biology of the qa gene cluster in neurospora crassa.an ensemble of genetic networks that describe how the model fungal system, neurospora crassa, utilizes quinic acid (qa) as a sole carbon source has been identified previously. a genetic network for qa metabolism involves the genes, qa-1f and qa-1s, that encode a transcriptional activator and repressor, respectively and structural genes, qa-2, qa-3, qa-4, qa-x, and qa-y. by a series of 4 separate and independent, model-guided, microarray experiments a total of 50 genes are identified as qa-respon ...201121695121
novel terpenoids of the fungus aspergillus insuetus isolated from the mediterranean sponge psammocinia sp. collected along the coast of israel.three novel meroterpenoids, insuetolides a-c (1-3) and four drimane sesquiterpenes, the new (e)-6-(4'-hydroxy-2'-butenoyl)-strobilactone a (4) and the known 2α, 9α, 11-trihydroxy-6-oxodrim-7-ene (5), strobilactone a (6) and (e,e)-6-(6',7'-dihydroxy-2',4'-octadienoyl)-strobilactone a (7), were isolated from the etoac extract of the culture medium of the marine-derived fungus aspergillus insuetus (oy-207), which was isolated from the mediterranean sponge psammocinia sp. the structures of the compo ...201121676619
transgenic nicotiana tabacum plants expressing a fungal copper transporter gene show enhanced acquisition of copper.the diets of two-thirds of the world's population are deficient in one or more essential elements and one of the approaches to enhance the levels of mineral elements in food crops is by developing plants with ability to accumulate them in edible parts. besides conventional methods, transgenic technology can be used for enhancing metal acquisition in plants. copper is an essential element, which is often deficient in human diet. with the objective of developing plants with improved copper acquisi ...201121671073
identification and characterization of genes required for cell-to-cell fusion in neurospora crassa.a screening procedure was used to identify cell fusion (hyphal anastomosis) mutants in the neurospora crassa single gene deletion library. mutants with alterations in 24 cell fusion genes required for cell fusion between conidial anastomosis tubes (cats) were identified and characterized. the cell fusion genes identified included 14 genes that are likely to function in signal transduction pathways needed for cell fusion to occur (mik-1, mek-1, mak-1, nrc-1, mek-2, mak-2, rac-1, pp2a, so/ham-1, h ...201121666072
fluorescent and bimolecular-fluorescent protein tagging of genes at their native loci in neurospora crassa using specialized double-joint pcr plasmids.the double-joint polymerase chain reaction (dj-pcr) is a technique that can be used to construct vectors for targeted genome integration without laborious subcloning steps. here we report the availability of plasmids that facilitate dj-pcr-based construction of neurospora crassa tagging vectors. these plasmids allow the creation of green or red fluorescent protein (gfp or rfp) tagging vectors for protein localization studies, as well as split-yellow fluorescent protein (yfp) tagging vectors for ...201121664475
crystal structure of the mid-piwi lobe of a eukaryotic argonaute protein.argonaute proteins (agos) are essential effectors in rna-mediated gene silencing pathways. they are characterized by a bilobal architecture, in which one lobe contains the n-terminal and paz domains and the other contains the mid and piwi domains. here, we present the first crystal structure of the mid-piwi lobe from a eukaryotic ago, the neurospora crassa qde-2 protein. compared to prokaryotic agos, the domain orientation is conserved, indicating a conserved mode of nucleic acid binding. the pi ...201121646546
probing the growth dynamics of neurospora crassa with microfluidic structures.despite occupying physically and chemically heterogeneous natural environments, the growth dynamics of filamentous fungi is typically studied on the surface of homogeneous laboratory media. fungal exploration and exploitation of complex natural environments requires optimal survival and growth strategies at the colony, hyphal, and intra hyphal level, with hyphal space-searching strategies playing a central role. we describe a new methodology for the characterisation and analysis of hyphal space- ...201121640314
architecture and development of the neurospora crassa hypha -- a model cell for polarized growth.neurospora crassa has been at the forefront of biological research from the early days of biochemical genetics to current progress being made in understanding gene and genetic network function. here, we discuss recent developments in analysis of the fundamental form of fungal growth, development and proliferation -- the hypha. understanding the establishment and maintenance of polarity, hyphal elongation, septation, branching and differentiation are at the core of current research. the advances ...201121640311
neurospora crassa mat a-2 and mat a-3 proteins weakly interact in the yeast two-hybrid system and affect yeast growth.mating-type genes control the entry into the sexual cycle, mating identity and sexual development in fungi. the mat a-2 and mat a-3 genes, present in the mat a idiomorph of the filamentous fungus neurospora crassa, are required for post-fertilization functions but are not essential for mating identity. their putative roles as transcription factors are based on the similarity of mat a-2 with the podospora anserina smr1 gene and an hmg motif present in the mat a-3 gene. in this work the yeast two- ...200921637691
a new diet for yeast to improve biofuel production.in 2010, our group announced the discovery of two cellodextrin transporter families from the cellulolytic fungus, neurospora crassa. furthermore, we demonstrated the utility of these transporters in the production of lignocellulosic biofuels. this discovery was made possible by a decision to systematically study cell wall degradation by n. crassa. the identified transport pathway has opened up a new way of thinking about microbial fermentation of hexoses as well as pentoses derived from plant ce ...201121637011
global warming, plant paraquat resistance, and light signal transduction through nucleoside diphosphate kinase as a paradigm for increasing food supply.light signal transduction was studied in extracts of mycelia of the fungus neurospora crassa, and the third internodes of dark-grown pisum sativum cv alaska. both processes increased the phosphorylation of nucleoside diphosphate kinase (ndpk). ndpk may function as a carrier of reduction equivalents, as it binds nadh, thereby providing electrons to transform singlet oxygen to superoxide by catalases (cat). as the c-termini of ndpk interact with cat which receive singlet oxygen, emitted from photo ...201121603975
characterization of neurospora crassa α-actinin.α-actinin, an actin-binding protein of the spectrin superfamily, is present in most eukaryotes except plants. it is composed of three domains: n-terminal ch-domains, c-terminal calcium-binding domain (with ef-hand motifs), and a central rod domain. we have cloned and expressed neurospora crassa α-actinin as gst and gfp fusion proteins for biochemical characterization and in vivo localization, respectively. the intracellular localization pattern of α-actinin suggests that this protein is intimate ...201121598047
phenotypic analysis of neurospora crassa gene deletion strains.phenotypic analysis of neurospora crassa knockout (ko) mutants was used as a vehicle to introduce students to laboratory research. the availability of gene deletion strains was the impetus for the development of a program designed to introduce beginning science students to basic microbiology, genetics, microscopy and beginning bioinformatics. the goal was to provide a research experience, acquire laboratory skills and phenotype hundreds of ko mutants. the data provided by the students was used t ...201121590422
high-throughput production of gene replacement mutants in neurospora crassa.the model filamentous fungus neurospora crassa has been the focus of functional genomics studies for the past several years. a high-throughput gene knockout procedure has been developed and used to generate mutants for more than two-thirds of the ∼10,000 annotated n. crassa genes. yeast recombinational cloning was incorporated as an efficient procedure to produce all knockout cassettes. n. crassa strains with the δmus-51 or δmus-52 deletion mutations were used as transformation recipients in ord ...201121590421
a predicted protein-protein interaction network of the filamentous fungus neurospora crassa.the filamentous fungus neurospora crassa is a leading model organism for circadian clock studies. computational identification of a protein-protein interaction (ppi) network (also known as an interactome) in n. crassa can provide new insights into the cellular functions of proteins. using two well-established bioinformatics methods (the interolog method and the domain interaction-based method), we predicted 27,588 ppis among 3006 n. crassa proteins. to the best of our knowledge, this is the firs ...201121584303
direct spectrophotometric assay of laccase using diazo derivatives of guaiacol.laccase (ec 1.10.3.2) is a widespread cuproenzyme able to oxidize various types of phenols and similar aromatic compounds through a one-electron transfer mechanism. the enzyme has already found its way into the market as a biocatalyst. because of its ability to be paired by electron mediators, the expectation for employing laccases in versatile processes is very high. there are a few spectrophotometric methods for assaying the laccase activity; however, all of them are based on the formation of ...201121545148
inactivation of snt2, a bah/phd-containing transcription factor, impairs pathogenicity and increases autophagosome abundance in fusarium oxysporum.the soil-borne, asexual fungus fusarium oxysporum f.sp. melonis (fom) is a causal agent of muskmelon wilt disease. the current study focused on the most virulent race of fom-race 1,2. the tagged mutant d122, generated by agrobacterium tumefaciens-mediated transformation, caused the delayed appearance of initial wilt disease symptoms, as well as a 75% reduction in pathogenicity. d122 was impaired in the gene product homologous to the snt2-like transcription factor of schizosaccharomyces pombe. in ...201121535351
involvement of a putative response regulator fgrrg-1 in osmotic stress response, fungicide resistance and virulence in fusarium graminearum.response regulator (rr) proteins are core elements of the high-osmolarity glycerol (hog) pathway, which plays an important role in the adaptation of fungi to a variety of environmental stresses. in this study, we constructed deletion mutants of two putative rr genes, fgrrg-1 and fgrrg-2, which are orthologues of neurospora crassa rrg-1 and rrg-2, respectively. the fgrrg-1 deletion mutant (δfgrrg1-6) showed increased sensitivity to osmotic stress mediated by nacl, kcl, sorbitol or glucose, and to ...201121535349
bulk segregant analysis followed by high-throughput sequencing reveals the neurospora cell cycle gene, ndc-1, to be allelic with the gene for ornithine decarboxylase, spe-1.with the advent of high-throughput dna sequencing, it is now straightforward and inexpensive to generate high-density small nucleotide polymorphism (snp) maps. here we combined high-throughput sequencing with bulk segregant analysis to expedite mutation mapping. the general map location of a mutation can be identified by a single backcross to a strain enriched in snps compared to a standard wild-type strain. bulk segregant analysis simultaneously increases the likelihood of determining the preci ...201121515825
heterochromatin is required for normal distribution of neurospora crassa cenh3.centromeres serve as platforms for the assembly of kinetochores and are essential for nuclear division. here we identified neurospora crassa centromeric dna by chromatin immunoprecipitation followed by high-throughput sequencing (chip-seq) of dna associated with tagged versions of the centromere foundation proteins cenh3 (cenp-a) and cen-c (cenp-c) and the kinetochore protein cen-t (cenp-t). on each chromosome we found an ∼150- to 300-kbp region of enrichment for all three proteins. these region ...201121505064
role of insulin in cr(vi)-mediated genotoxicity in neurospora crassa.chromium (iii) is an insulinomimetic agent whose biological and/or environmental availability is frequently in the form of cr(vi), which is known to be toxic. wall-less mutant of neurospora crassa (fgsc stock no. 4761) is known to possess insulin receptor in its cell membrane and hence is a good model for cr toxicity studies. this study explores the toxicity of cr(vi) and the possible consequences on simultaneous exposure to insulin in n. crassa.201121488912
use of 1h nuclear magnetic resonance to measure intracellular metabolite levels during growth and asexual sporulation in neurospora crassa.conidiation is an asexual sporulation pathway that is a response to adverse conditions and is the main mode of dispersal utilized by filamentous fungal pathogens for reestablishment in a more favorable environment. heterotrimeric g proteins (consisting of α, β, and γ subunits) have been shown to regulate conidiation in diverse fungi. previous work has demonstrated that all three of the gα subunits in the filamentous fungus neurospora crassa affect the accumulation of mass on poor carbon sources ...201121460191
phosphorylations: making the neurosporacrassa circadian clock tick.various post-translational modifications have been identified that play a role in the function of circadian clocks. among these, phosphorylation has been investigated extensively. it was shown that phosphorylation influences half-life, subcellular localisation, transcriptional activity and conformation of clock components over the course of a circadian day. recent observations also indicate that time-of-day specific sequential phosphorylation of the neurospora crassa clock protein frequency is c ...201121453704
light input and processing in the circadian clock of neurospora.circadian clocks are endogenous oscillators that use zeitgebers as environmental cues to synchronise with the exogenous day-night cycle. the role of light as a zeitgeber has been investigated intensively to date. in neurospora crassa the transcription factor white collar complex (wcc) is directly activated by light, which resets the clock. in addition, a hierarchical cascade of transcription factors activates the light-induced expression of hundreds of genes. disturbance of the clock during the ...201121453703
nmr structure of the c-terminal domain of a tyrosyl-trna synthetase that functions in group i intron splicing.the mitochondrial tyrosyl-trna synthetases (mt tyrrss) of pezizomycotina fungi are bifunctional proteins that aminoacylate mitochondrial trna(tyr) and are structure-stabilizing splicing cofactors for group i introns. studies with the neurospora crassa synthetase (cyt-18 protein) showed that splicing activity is dependent upon pezizomycotina-specific structural adaptations that form a distinct group i intron-binding site in the n-terminal catalytic domain. although cyt-18's c-terminal domain also ...201121438536
neurosporaside, a tetraglycosylated sphingolipid from neurospora crassa.the new tetraglycosylceramide neurosporaside (1a) has been isolated from the fungus neurospora crassa. neurosporaside is a tetraglycosylated glycosphingolipid characterized by a sugar chain unprecedented among natural glycoconjugates. the structure of neurosporaside was elucidated by extensive spectroscopic analysis and microscale degradation analysis, which allowed full structure elucidation using less than 1 mg of compound.201121425845
problem of interactions between intracellular structures during the neurospora crassa tip growth. 201121369903
nmr solution structure of a cyanovirin homolog from wheat head blight fungus.members of the cyanovirin-n homolog (cvnh) lectin family are found in bacteria, fungi and plants. as part of our ongoing work on cvnh structure-function studies, we determined the high-resolution nmr solution structure of the homolog from the wheat head blight disease causing ascomycetous fungus gibberella zeae (or fusarium graminearum), hereafter called gzcvnh. like cyanovirin-n (cv-n), gzcvnh comprises two tandem sequence repeats and the protein sequence exhibits 30% identity with cv-n. the ov ...201121365681
mid1, a mechanosensitive calcium ion channel, affects growth, development, and ascospore discharge in the filamentous fungus gibberella zeae.the role of mid1, a stretch-activated ion channel capable of being permeated by calcium, in ascospore development and forcible discharge from asci was examined in the pathogenic fungus gibberella zeae (anamorph fusarium graminearum). the ?mid1 mutants exhibited a >12-fold reduction in ascospore discharge activity and produced predominately abnormal two-celled ascospores with constricted and fragile septae. the vegetative growth rate of the mutants was ~50% of the wild-type rate, and production o ...201121357477
a reporter for dsrna response in neurospora crassa.in the filamentous fungus neurospora, the production of dsrna can elicit a dsrna-induced transcriptional response similar to the interferon response in vertebrates. however, how fungi sense the expression of dsrna and activate gene expression is unknown. in this study, we established a dsrna response reporter system in neurospora crassa. using the dsrna-activated rna-dependent rna polymerase gene rrp-3 promoter, we created an expression construct (prrp-3::myc-al-1) and introduced it into al-1(ko ...201121354420
combined use of paracoccidioides brasiliensis recombinant rpb27 and rpb40 antigens in an enzyme-linked immunosorbent assay for immunodiagnosis of paracoccidioidomycosis.paracoccidioidomycosis (pcm) is one of the most important endemic mycoses in latin america; it's usually diagnosed by observation and/or isolation of the etiologic agent, paracoccidioides brasiliensis, as well as by a variety of immunological methods, such as complement fixation and immunodiffusion. although these approaches are useful, historically their sensitivity and specificity have often been compromised by the use of complex mixtures of undefined antigens. the use of combinations of purif ...201121354162
lilbid-mass spectrometry of the mitochondrial preprotein translocase tom.in the present work we applied a novel mass spectrometry method termed laser-induced liquid bead ion desorption mass spectrometry (lilbid-ms) to the outer mitochondrial membrane protein translocon tom to analyze its subunit composition and stoichiometry. with tom core complex, purified at high ph, we demonstrate that a tom core complex of neurospora crassa is composed of at least two tom40 and tom22 molecules, respectively, and more than five small tom subunits between 5.5 and 6.4 kda. we show t ...201021339618
role of four calcium transport proteins, encoded by nca-1, nca-2, nca-3, and cax, in maintaining intracellular calcium levels in neurospora crassa.we have examined the distribution of calcium in neurospora crassa and investigated the role of four predicted calcium transport proteins. the results of cell fractionation experiments showed 4% of cellular calcium in mitochondria, approximately 11% in a dense vacuolar fraction, 40% in an insoluble form that copurifies with microsomes, and 40% in a high-speed supernatant, presumably from large vacuoles that had broken. strains lacking nca-1, a serca-type ca(2+)-atpase, or nca-3, a pmc-type ca(2+) ...201121335528
aspergillus nidulans transcription factor atfa interacts with the mapk saka to regulate general stress responses, development and spore functions.fungi utilize a phosphorelay system coupled to a map kinase module for sensing and processing environmental signals. in aspergillus nidulans, response regulator sska transmits osmotic and oxidative stress signals to the stress mapk (sapk) saka. using a genetic approach together with gfp tagging and molecular bifluorescence we show that saka and atf/creb transcription factor atfa define a general stress-signalling pathway that plays differential roles in oxidative stress responses during growth a ...201121320182
traffic of chitin synthase 1 (chs-1) to the spitzenkörper and developing septa in hyphae of neurospora crassa: actin dependence and evidence of distinct microvesicle populations.we describe the subcellular location of chitin synthase 1 (chs-1), one of seven chitin synthases in neurospora crassa. laser scanning confocal microscopy of growing hyphae showed chs-1-green fluorescent protein (gfp) localized conspicuously in regions of active wall synthesis, namely, the core of the spitzenkörper (spk), the apical cell surface, and developing septa. it was also present in numerous fine particles throughout the cytoplasm plus some large vacuoles in distal hyphal regions. althoug ...201121296914
carrefour mme. gras: a wild-isolated neurospora crassa strain that suppresses meiotic silencing by unpaired dna and uncovers a novel ascospore stability defect.ordinarily, rip-induced erg-3 mutant neurospora crassa ascospores and their erg(+) siblings do not differ in stability during long-term storage. consequently, the frequency of rip-induced erg-3 mutants remains about constant regardless of the time that has elapsed between ascospore harvest and germination. we found, however, that rip-induced erg-3 mutants were apparently selectively lost with time from among the ascospores stored from a cross with the wild-isolated carrefour mme. gras strain fro ...201121295150
translocations used to generate chromosome segment duplications in neurospora can disrupt genes and create novel open reading frames.in neurospora crassa, crosses between normal sequence strains and strains bearing some translocations can yield progeny bearing a duplication (dp) of the translocated chromosome segment. here, 30 breakpoint junction sequences of 12 dp-generating translocations were determined. the breakpoints disrupted 13 genes (including predicted genes), and created 10 novel open reading frames. insertion of sequences from lg iii into lg i as translocation t(uk8-18) disrupts the eat-3 gene, which is the orthol ...201021289436
population genomics and local adaptation in wild isolates of a model microbial eukaryote.elucidating the connection between genotype, phenotype, and adaptation in wild populations is fundamental to the study of evolutionary biology, yet it remains an elusive goal, particularly for microscopic taxa, which comprise the majority of life. even for microbes that can be reliably found in the wild, defining the boundaries of their populations and discovering ecologically relevant phenotypes has proved extremely difficult. here, we have circumvented these issues in the microbial eukaryote n ...201121282627
identification of macrodomain proteins as novel o-acetyl-adp-ribose deacetylases.sirtuins are a family of protein lysine deacetylases, which regulate gene silencing, metabolism, life span, and chromatin structure. sirtuins utilize nad(+) to deacetylate proteins, yielding o-acetyl-adp-ribose (oaadpr) as a reaction product. the macrodomain is a ubiquitous protein module known to bind adp-ribose derivatives, which diverged through evolution to support many different protein functions and pathways. the observation that some sirtuins and macrodomains are physically linked as fusi ...201121257746
expression of fungal diacylglycerol acyltransferase2 genes to increase kernel oil in maize.maize (zea mays) oil has high value but is only about 4% of the grain by weight. to increase kernel oil content, fungal diacylglycerol acyltransferase2 (dgat2) genes from umbelopsis (formerly mortierella) ramanniana and neurospora crassa were introduced into maize using an embryo-enhanced promoter. the protein encoded by the n. crassa gene was longer than that of u. ramanniana. it included 353 amino acids that aligned to the u. ramanniana dgat2a protein and a 243-amino acid sequence at the amino ...201121245192
role of cell wall bound calcium in neurospora crassa.cell wall bound calcium constitutes a significant fraction (25%) of total mycelia calcium in neurospora crassa. wall bound calcium increases as a function of growth and calcium concentration, while cell wall bound calcium decreases in ca-free medium. removal of wall bound calcium causes its rapid replacement from intracellular pool, inhibited by verapamil, nifedipine, concanamycin a, and wortmanin in a vacuolar mutant (vma-5), but is unaffected by trifluoropyrazine, and calmidizoluim in a calcin ...201121237628
acon-3, the neurospora crassa ortholog of the developmental modifier, meda, complements the conidiation defect of the aspergillus nidulans mutant.aspergillus nidulans and neurospora crassa are ascomycetes that produce asexual spores through morphologically distinct processes. meda, a protein with unknown function, is required for normal asexual and sexual development in a. nidulans. we determined that the n. crassa ortholog of meda is acon-3, a gene required for early conidiophore development and female fertility. to test hypotheses about the evolutionary origins of asexual development in distinct fungal lineages it is important to unders ...201121220038
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