parp is involved in replicative aging in neurospora crassa.modification of proteins by the addition of poly(adp-ribose) is carried out by poly(adp-ribose) polymerases (parps). parps have been implicated in a wide range of biological processes in eukaryotes, but no universal function has been established. a study of the aspergillus nidulans parp ortholog (prpa) revealed that the protein is essential and involved in dna repair, reminiscent of findings using mammalian systems. we found that a neurospora parp orthologue (npo) is dispensable for cell surviva ...201020045739
adenylyl cyclase regulates heavy metal sensitivity, bikaverin production and plant tissue colonization in fusarium proliferatum.a homologue of the adenylyl cyclase (ac) gene of neurospora crassa, named fpacy1 was cloned from the genomic library of fusarium proliferatum item 2287 by screening the library with a dna fragment amplified by using pcr primers designed from conserved sequences of the catalytic domain of ac genes from other fungi. the deduced fpacy1 protein had 53-77% identity with the ac proteins of other fungi. deltafpacy1 mutants obtained by targeted gene disruption showed retarded vegetative growth, increase ...201020082366
[phylogenetic analysis and detection of selective pressure on the cvnh domains].cyanovirin-n (cv-n) is a novel protein with broad-spectrum antiviral activity. its homologs constitute a protein family known as cvnh (cyanovirin-n homology), which possess the evolutionarily conserved anti-hiv (human immunodeficiency virus) domain. in this study, more details about the patchy organism distribution of cvnh domain were explored by reconstructing gene trees. duplicated cvnh sequences were also identified in a wide range of species including aspergillus niger, neosartorya fischeri ...201020085891
f-actin dynamics in neurospora crassa.this study demonstrates the utility of lifeact for the investigation of actin dynamics in neurospora crassa and also represents the first report of simultaneous live-cell imaging of the actin and microtubule cytoskeletons in filamentous fungi. lifeact is a 17-amino-acid peptide derived from the nonessential saccharomyces cerevisiae actin-binding protein abp140p. fused to green fluorescent protein (gfp) or red fluorescent protein (tagrfp), lifeact allowed live-cell imaging of actin patches, cable ...201020139238
the role of paaac1 encoding a mitochondrial adp/atp carrier in the biosynthesis of extracellular glycolipids, mannosylerythritol lipids, in the basidiomycetous yeast pseudozyma antarctica.pseudozyma antarctica produces large amounts of the glycolipid biosurfactants known as mannosylerythritol lipids (mel), which show not only excellent surface-active properties but also versatile biochemical actions. a gene homologous with a mitochondrial adp/atp carrier was dominantly expressed in p. antarctica under mel-producing conditions on the basis of previous gene expression analysis. the gene encoding the mitochondrial adp/atp carrier of p. antarctica (paaac1) contained a putative open r ...201020146402
a single mutation in the first transmembrane domain of yeast cox2 enables its allotopic expression.during the course of evolution, a massive reduction of the mitochondrial genome content occurred that was associated with transfer of a large number of genes to the nucleus. to further characterize factors that control the mitochondrial gene transfer/retention process, we have investigated the barriers to transfer of yeast cox2, a mitochondrial gene coding for a subunit of cytochrome c oxidase complex. nuclear-recoded saccharomyces cerevisiae cox2 fused at the amino terminus to various alternati ...201020194738
septum formation is regulated by the rho4-specific exchange factors bud3 and rgf3 and by the landmark protein bud4 in neurospora crassa.rho gtpases have multiple, yet poorly defined functions during cytokinesis. by screening a neurospora crassa knock-out collection for rho guanine nucleotide exchange factor (gef) mutants that phenocopy rho-4 defects (i.e. lack of septa, slow growth, abnormal branching and cytoplasmic leakage), we identified two strains defective in homologues of bud3p and rgf3 of budding and fission yeast respectively. the function of these proteins as rho4-specific gefs was determined by in vitro assays. in viv ...201020199606
nuclear dynamics, mitosis, and the cytoskeleton during the early stages of colony initiation in neurospora crassa.neurospora crassa macroconidia form germ tubes that are involved in colony establishment and conidial anastomosis tubes (cats) that fuse to form interconnected networks of conidial germlings. nuclear and cytoskeletal behaviors were analyzed in macroconidia, germ tubes, and cats in strains that expressed fluorescently labeled proteins. heterokaryons formed by cat fusion provided a rapid method for the imaging of multiple labeled fusion proteins and minimized the potential risk of overexpression a ...201020207852
high-throughput construction of gene deletion cassettes for generation of neurospora crassa knockout strains.the availability of complete genome sequences for a number of biologically important fungi has become an important resource for fungal research communities. however, the functions of many open reading frames (orfs) identified through annotation of whole genome sequences have yet to be determined. the disruption of orfs is a practical method for loss-of-function gene analyses in fungi that are amenable to transformation. unfortunately, the construction of knockout cassettes using traditional dige ...201020238259
visualization of f-actin localization and dynamics with live cell markers in neurospora crassa.filamentous actin (f-actin) plays essential roles in filamentous fungi, as in all other eukaryotes, in a wide variety of cellular processes including cell growth, intracellular motility, and cytokinesis. we visualized f-actin organization and dynamics in living neurospora crassa cells via confocal microscopy of growing hyphae expressing gfp fusions with homologues of the actin-binding proteins fimbrin (fim) and tropomyosin (tpm-1), a subunit of the arp2/3 complex (arp-3) and a recently developed ...201020302965
genetic and molecular characterization of a cryptochrome from the filamentous fungus neurospora plants and animals, cryptochromes function as either photoreceptors or circadian clock components. we have examined the cryptochrome from the filamentous fungus neurospora crassa and demonstrate that neurospora cry encodes a dash-type cryptochrome that appears capable of binding flavin adenine dinucleotide (fad) and methenyltetrahydrofolate (mthf). the cry transcript and cry protein levels are strongly induced by blue light in a wc-1-dependent manner, and cry transcript is circadianly regulat ...201020305004
roles of the mdm10, tom7, mdm12, and mmm1 proteins in the assembly of mitochondrial outer membrane proteins in neurospora crassa.the mdm10, mdm12, and mmm1 proteins have been implicated in several mitochondrial functions including mitochondrial distribution and morphology, assembly of beta-barrel proteins such as tom40 and porin, association of mitochondria and endoplasmic reticulum, and maintaining lipid composition of mitochondrial membranes. here we show that loss of any of these three proteins in neurospora crassa results in the formation of large mitochondrial tubules and reduces the assembly of porin and tom40 into ...201020335503
an ap-1-like transcription factor, nap-1, regulates expression of the glutathione s-transferase and nadh:flavin oxidoreductase genes in neurospora crassa.ap-1-like transcription factors play crucial roles in oxidative stress responses in yeast and filamentous fungi. the deletion of an ap-1-like transcription factor gene, nap-1, in neurospora crassa slightly increased its sensitivity to oxidative stressors, including menadione. microarray and quantitative real-time reverse transcriptase-pcr analyses were employed to identify menadione-inducible genes (migs) and the roles of nap-1 in their regulation. n. crassa migs include three putative glutathio ...201020378991
the fungal genetics stock center: a repository for 50 years of fungal genetics research.the fungal genetics stock center (fgsc) was established in 1960 to ensure that important strains used in early genetics research were available to subsequent generations of fungal geneticists. originally, only mutant strains were held. at present, any organism that has had its genome sequenced is a genetic system and so the fgsc has added many new organisms. the fgsc is well integrated in its core community and, as research came to depend on cloned genes, vectors and gene libraries, the fgsc inc ...201020413916
rotenone enhances the antifungal properties of staurosporine.we studied staurosporine-induced cell death in the filamentous fungus neurospora crassa. the generation of reactive oxygen species during the process appears to be an important signaling event, since addition of the antioxidant glutathione prevents the effects of staurosporine on fungal growth. selected mutants with mutations in respiratory chain complex i are extremely sensitive to the drug, stressing the involvement of complex i in programmed cell death. following this finding, we determined t ...201020435699
extensive and varied modifications in histone h2b of wild-type and histone deacetylase 1 mutant neurospora crassa.dna methylation is deficient in a histone deacetylase 1 (hda1) mutant (hda-1) strain of neurospora crassa with inactivated histone deacetylase 1. difference two-dimensional (2d) gels identified the primary histone deacetylase 1 target as histone h2b. acetylation was identified by lc-ms/ms at five different lysines in wild-type h2b and at 11 lysines in hda-1 h2b, suggesting neurospora h2b is a complex combination of different acetylated species. individual 2d gel spots were shifted by single lysi ...201020462202
photobiology in the zygomycota: multiple photoreceptor genes for complex responses to light.light is an environmental signal that modulates many aspects of the biology of zygomycete fungi. light regulation has been investigated in the zygomycetes phycomyces blakesleeanus, mucor circinelloides and pilobolus crystallinus. examples of light regulation include the phototropism of the fruiting bodies, the regulation of the development of reproductive structures, and the activation of the biosynthesis of β-carotene. in fungi blue light is perceived by proteins homologous to wc-1, a neurospor ...201020466063
regulation of fruiting body photomorphogenesis in coprinopsis cinerea.the agaricomycete (homobasidiomycete) coprinopsis cinerea has been used as a model to study the molecular mechanism for photomorphogenesis. molecular genetic analyses of mutants defective in fruiting body (mushroom) photomorphogenesis of c. cinerea identified two genes, dst1 and dst2. dst1 encodes a homolog of wc-1, a fungal blue-light photoreceptor first identified in neurospora crassa, while dst2 encodes a novel protein with a putative flavin adenine dinucleotide (fad)-binding-4 domain. in add ...201020471485
ferrochelatase is a conserved downstream target of the blue light-sensing white collar complex in fungi.light is a universal signal perceived by organisms, including fungi, in which light regulates common and unique biological processes depending on the species. previous research has established that conserved proteins, originally called white collar 1 and 2 from the ascomycete neurospora crassa, regulate uv/blue light sensing. homologous proteins function in distant relatives of n. crassa, including the basidiomycetes and zygomycetes, which diverged as long as a billion years ago. here we conduct ...201020488877
expression of a neurospora crassa zinc transporter gene in transgenic nicotiana tabacum enhances plant zinc accumulation without co-transport of cadmium.zinc (zn) is an essential micronutrient required for growth and development of all organisms. deficiency of zn in humans is widespread, affecting 25% of world population and efforts are underway to develop crop plants with high levels of zn in their edible parts. when strategies for enhancing zn in crop plants are designed, it is essential to exclude cadmium (cd), a toxic analogue of zn. in the present work, a high affinity and high specificity zinc transporter gene (tzn1) from neurospora crassa ...201020492552
the global nitrogen regulator, fnr1, regulates fungal nutrition-genes and fitness during fusarium oxysporum pathogenesis.summary fusarium oxysporum is a soil-borne pathogen that infects plants through the roots and uses the vascular system for host ingress. specialized for this route of infection, f. oxysporum is able to adapt to the scarce nutrient environment in the xylem vessels. here we report the cloning of the f. oxysporum global nitrogen regulator, fnr1, and show that it is one of the determinants for fungal fitness during in planta growth. the fnr1 gene has a single conserved gata-type zinc finger domain a ...200620507463
meiotic regulators ndt80 and ime2 have different roles in saccharomyces and neurospora.meiosis is a highly regulated process in eukaryotic species. the filamentous fungus neurospora crassa has been shown to be missing homologs of a number of meiotic initiation genes conserved in saccharomyces cerevisiae, but has three homologs of the well-characterized middle meiotic transcriptional regulator ndt80. in this study, we evaluated the role of all three ndt80 homologs in the formation of female reproductive structures, sexual development, and meiosis. we found that none of the ndt80 ho ...201020519745
the ham-5, rcm-1 and rco-1 genes regulate hyphal fusion in neurospora crassa.mutants of neurospora crassa unable to participate in vegetative hyphal fusion (anastomosis) were isolated and characterized. from this analysis, three genes, rcm-1, rco-1 and ham-5, were identified and shown to be required for hyphal fusion. the rcm-1 and rco-1 genes are homologues of the saccharomyces cerevisiae ssn6 and tup1 genes, which encode a dimeric transcription factor in yeast. we demonstrate that in n. crassa the rcm-1 and rco-1 genes are required for hyphal fusion and normal hyphal m ...201020522492
functional conservation of mei4 for meiotic dna double-strand break formation from yeasts to mice.meiotic recombination is initiated by the programmed induction of dna double-strand breaks (dsbs) catalyzed by the evolutionarily conserved spo11 protein. studies in yeast have shown that dsb formation requires several other proteins, the role and conservation of which remain unknown. here we show that two of these saccharomyces cerevisiae proteins, mei4 and rec114, are evolutionarily conserved in most eukaryotes. mei4(-/-) mice are deficient in meiotic dsb formation, thus showing the functional ...201020551173
dual roles for the mss116 cofactor during splicing of the ai5γ group ii intron.the autocatalytic group ii intron ai5γ from saccharomyces cerevisiae self-splices under high-salt conditions in vitro, but requires the assistance of the dead-box protein mss116 in vivo and under near-physiological conditions in vitro. here, we show that mss116 influences the folding mechanism in several ways. by comparing intron precursor rnas with long (∼300 nt) and short (∼20 nt) exons, we observe that long exon sequences are a major obstacle for self-splicing in vitro. kinetic analysis indic ...201020554854
[spectrophometric analysis of volatile compounds in microorganisms].a simple modification of a spectrophometric method was proposed for the rapid detection of microorganisms based on their ability either to excrete or to absorb volatile compounds. the method provides the possibility of contactless control for bacterial growth at a concentration above 10(7) cells/ml. in addition, the method allows discriminating mutants of the fungus neurospora crassa defective in the nitrogen metabolism from the wild type strains. it is likely that nitrite reductase and nitrate ...201020586286
the mitogen-activated protein kinase kinase bos5 is involved in regulating vegetative differentiation and virulence in botrytis cinerea.we present a characterization of bos5 from botrytis cinerea, a gene that encodes a mitogen-activated protein kinase kinase (mapkk), which is homologous to os-5 of neurospora crassa. the bos5 gene deletion mutant exhibited reduced vegetative growth and strongly impaired conidiation. the mutant also exhibited increased sensitivity to the dicarboximide fungicide iprodione and to osmotic stress mediated by nacl or kcl. western-blot analysis showed that the bcsak1 protein, the putative downstream com ...201020595070
genes encoding a striatin-like protein (ham-3) and a forkhead associated protein (ham-4) are required for hyphal fusion in neurospora crassa.cell-cell fusion during fertilization and between somatic cells is an integral process in eukaryotic development. in neurospora crassa, the hyphal anastomosis mutant, ham-2, fails to undergo somatic fusion. in both humans and saccharomyces cerevisiae, homologs of ham-2 are found in protein complexes that include homologs to a striatin-like protein and a forkhead-associated (fha) protein. we identified a striatin (ham-3) gene and a fha domain (ham-4) gene in n. crassa; strains containing mutation ...201020601042
the antifungal activity of the penicillium chrysogenum protein paf disrupts calcium homeostasis in neurospora crassa.the antifungal protein paf from penicillium chrysogenum exhibits growth-inhibitory activity against a broad range of filamentous fungi. evidence from this study suggests that disruption of ca(2+) signaling/homeostasis plays an important role in the mechanistic basis of paf as a growth inhibitor. supplementation of the growth medium with high ca(2+) concentrations counteracted paf toxicity toward paf-sensitive molds. by using a transgenic neurospora crassa strain expressing codon-optimized aequor ...201020622001
aspergillus oryzae type iii polyketide synthase csya is involved in the biosynthesis of 3,5-dihydroxybenzoic a novel superfamily of type iii polyketide synthases in microbes, four genes csya, csyb, csyc, and csyd, were found in the genome of aspergillus oryzae, an industrially important filamentous fungus. in order to analyze their functions, we carried out the overexpression of csya under the control of alpha-amylase promoter in a. oryzae and identified 3,5-dihydroxybenzoic acid (dhba) as the major product. feeding experiments using (13)c-labeled acetates confirmed that the acetate labeling pattern ...201020630753
confining euchromatin/heterochromatin territory: jumonji crosses the line.heterochromatin is typically highly condensed, gene-poor, and transcriptionally silent, whereas euchromatin is less condensed, gene-rich, and more accessible to transcription. besides acting as a graveyard for selfish mobile dna repeats, heterochromatin contributes to important biological functions, such as chromosome segregation during cell division. multiple features of heterochromatin-including the presence or absence of specific histone modifications, dna methylation, and small rnas-have bee ...201020634313
genome-wide analysis of microsatellite sequence in seven filamentous fungi.abundance of microsatellites with repeated unit lengths of 1-6 base pairs in seven fungi: aspergillus nidulans, coprinus cinereus, cryptococcus neoformans (serotype a), fusarium graminearum, magnaporthe grisea, neurospora crassa and ustilago maydis were investigated on genomic scale. the results showed that each species has its specific profile for different types and different motifs of ssr loci. ascomycetes fungi m. grisea, n. crassa and basidiomycete fungus u. maydis adopt much more microsate ...200920640828
a comparative study of activity and apparent inhibition of fungal β-glucosidases.β-glucosidases (bgs) from aspergillus fumigates, aspergillus niger, aspergillus oryzae, chaetomium globosum, emericella nidulans, magnaporthe grisea, neurospora crassa, and penicillium brasilianum were purified to homogeneity, and analyzed by isothermal titration calorimetry with respect to their hydrolytic activity and its sensitivity to glucose (product) using cellobiose as substrate. global non-linear regression of several reactions, with or without added glucose, to a product inhibition equa ...201020677177
rna interference pathways in filamentous fungi.rna interference is a conserved homology-dependent post-transcriptional/transcriptional gene silencing mechanism in eukaryotes. the filamentous fungus neurospora crassa is one of the first organisms used for rnai studies. quelling and meiotic silencing by unpaired dna are two rnai-related phenomena discovered in neurospora, and their characterizations have contributed significantly to our understanding of rnai mechanisms in eukaryotes. a type of dna damage-induced small rna, microrna-like small ...201020680389
cazymes analysis toolkit (cat): web service for searching and analyzing carbohydrate-active enzymes in a newly sequenced organism using cazy database.the carbohydrate-active enzyme (cazy) database provides a rich set of manually annotated enzymes that degrade, modify, or create glycosidic bonds. despite rich and invaluable information stored in the database, software tools utilizing this information for annotation of newly sequenced genomes by cazy families are limited. we have employed two annotation approaches to fill the gap between manually curated high-quality protein sequences collected in the cazy database and the growing number of oth ...201020696711
expression of cellobiose dehydrogenase from neurospora crassa in pichia pastoris and its purification and characterization.a gene encoding cellobiose dehydrogenase (cdh) from neurospora crassa strain fgsc 2489 has been cloned and expressed in the heterologous host pichia pastoris, under the control of the aox1 methanol inducible promoter. recombinant cdh without the native signal sequence and fused with a his(6)-tag (rnc-cdh1) was successfully expressed and secreted. rnc-cdh1 was produced at the level of 652 iu/l after 2 days of cultivation in the induction medium. the his(6)-tagged rnc-cdh1 was purified through a o ...201120709172
discovery and characterization of novel d-xylose-specific transporters from neurospora crassa and pichia stipitis.saccharomyces cerevisiae is considered one of the most promising organisms for ethanol production from lignocellulosic feedstock. unfortunately, pentose sugars, which comprise up to 30% of lignocellulose, cannot be utilized by wild type s. cerevisiae. heterologous pathways were introduced into s. cerevisiae to enable utilization of d-xylose, the most abundant pentose sugar. however, the resulting recombinant s. cerevisiae strains exhibited a slow growth rate and poor sugar utilization efficiency ...201020714641
multi-targeted priming for genome-wide gene expression assays.complementary approaches to assaying global gene expression are needed to assess gene expression in regions that are poorly assayed by current methodologies. a key component of nearly all gene expression assays is the reverse transcription of transcribed sequences that has traditionally been performed by priming the poly-a tails on many of the transcribed genes in eukaryotes with oligo-dt, or by priming rna indiscriminately with random hexamers. we designed an algorithm to find common sequence m ...201020716356
identification of pex33, a novel component of the peroxisomal docking complex in the filamentous fungus neurospora crassa.the docking complex of peroxisomal matrix protein import is composed of pex13 and pex14 in all species analyzed so far, whereas only yeast appears to possess an additional component, pex17. in this report we isolated pex14 complexes of neurospora crassa. among the complex constituents, one protein designated as pex33 possessed homology to pex14 but only in a short n-terminal domain. the pex14/pex33 interaction was verified by means of two-hybrid analysis. moreover, pex33 was shown to interact wi ...201020728240
expression and characterization of the neurospora crassa endoglucanase gh5-1.filamentous fungi secrete a wide range of enzymes, including cellulases and hemicellulases, with potential applications in the production of lignocellulosic biofuels. of the cellulolytic fungi, hypocrea jecorina (anamorph trichoderma reesei) is the best characterized in terms of cellulose degradation, but other cellulolytic fungi, such as the model filamentous fungus neurospora crassa, can serve a crucial role in building our knowledge about the fungal response to biomass due to the many molecul ...201120826217
cellodextrin transport in yeast for improved biofuel production.fungal degradation of plant biomass may provide insights for improving cellulosic biofuel production. we show that the model cellulolytic fungus neurospora crassa relies on a high-affinity cellodextrin transport system for rapid growth on cellulose. reconstitution of the n. crassa cellodextrin transport system in saccharomyces cerevisiae promotes efficient growth of this yeast on cellodextrins. in simultaneous saccharification and fermentation experiments, the engineered yeast strains more rapid ...201020829451
genome mining for the discovery of new nitrilases in filamentous fungi.purpose of work: our aim is to describe new fungal nitrilases whose sequences were published but whose catalytic properties were unknown. we adapted for expression in e. coli three of the genes and confirmed that the enzymes acted on organic nitriles. the genome mining approach was used to search for nitrilases in filamentous fungi. synthetic genes encoding nitrilases in aspergillus niger, gibberella moniliformis and neurospora crassa were expressed in escherichia coli. this is the first heterol ...201020882316
archazolid a binds to the equatorial region of the c-ring of the vacuolar h+-atpase.the macrolactone archazolid is a novel, highly specific v-atpase inhibitor with an ic(50) value in the low nanomolar range. the binding site of archazolid is presumed to overlap with the binding site of the established plecomacrolide v-atpase inhibitors bafilomycin and concanamycin in subunit c of the membrane-integral v(o) complex. using a semi-synthetic derivative of archazolid for photoaffinity labeling of the v(1)v(o) holoenzyme we confirmed binding of archazolid to the v(o) subunit c. for t ...201020884613
fungal development and the cop9 signalosome.the conserved cop9 signalosome (csn) multiprotein complex is located at the interface between cellular signaling, protein modification, life span and the development of multicellular organisms. csn is required for light-controlled responses in filamentous fungi. this includes the circadian rhythm of neurospora crassa or the repression of sexual development by light in aspergillus nidulans. in contrast to plants and animals, csn is not essential for fungal viability. therefore fungi are suitable ...201020934903
rhythmic conidiation in the blue-light fungus trichoderma pleuroticola.trichoderma species conidiate in response to blue light, however, unlike in the blue-light model fungus neurospora crassa, conidiation in trichoderma spp. has been considered to be non-circadian. in this study we uncovered evidence for circadian conidiation in trichoderma pleuroticola and identified orthologues of the key n. crassa clock components, wc-1 (blr-1) and frq.201020943132
genome-wide prediction of g protein-coupled receptors in verticillium spp.g protein-coupled receptors (gpcrs) are critical factors in regulating morphogenesis, mating, infection and virulence in fungi. in this study, various computational strategies were applied to identify gpcr-like proteins from the genomes of both verticillium dahliae and verticillium albo-atrum. the putative gpcrs were distributed over 13 classes, and significantly, three of those represented novel classes of gpcr-like proteins in fungi. the three novel gpcrs had high levels of identity to their c ...201020943146
a circadian rhythm regulating hyphal melanization in cercospora kikuchii.many metabolic and developmental processes in fungi are controlled by biological rhythms. circadian rhythms approximate a daily (24 h) cycle and have been thoroughly studied in the model fungus, neurospora crassa. however relatively few examples of true circadian rhythms have been documented among other filamentous fungi. in this study we describe a circadian rhythm underlying hyphal melanization in cercospora kikuchii, an important pathogen of soybean. after growth in light or light : dark cycl ...201020943572
live-cell imaging of conidial fusion in the bean pathogen, colletotrichum lindemuthianum.fusion of conidia and conidial germlings by means of conidial anastomosis tubes (cats) is a common phenomenon in filamentous fungi, including many plant pathogens. it has a number of different roles, and has been speculated to facilitate parasexual recombination and horizontal gene transfer between species. the bean pathogen colletotrichum lindemuthianum naturally undergoes cat fusion on the host surface and within asexual fruiting bodies in anthracnose lesions on its host. it has not been previ ...201020965055
characterization of a thioredoxin (bbtrx) from the entomopathogenic fungus beauveria bassiana and its expression in response to thermal stress.a thioredoxin (bbtrx) was identified from the entomopathogenic fungus beauveria bassiana. the cloned nucleotide sequence consisted of a 423-bp open reading frame encoding a 141-amino-acid thioredoxin, a 1011-bp 5' region, and a 419-bp 3' region. the deduced protein sequence of bbtrx, including a common 95-amino-acid conserved domain and a unique 46-amino-acid carboxy terminal region, was similar (≤38% identity) to that of other thioredoxins and phylogenetically closest to that from neurospora cr ...201021076484
exploring the bzip transcription factor regulatory network in neurospora crassa.transcription factors (tfs) are key nodes of regulatory networks in eukaryotic organisms, including filamentous fungi such as neurospora crassa. the 178 predicted dna-binding tfs in n. crassa are distributed primarily among six gene families, which represent an ancient expansion in filamentous ascomycete genomes; 98 tf genes show detectable expression levels during vegetative growth of n. crassa, including 35 that show a significant difference in expression level between hyphae at the periphery ...201021081763
conserved components, but distinct mechanisms for the placement and assembly of the cell division machinery in unicellular and filamentous ascomycetes.cytokinesis is essential for cell proliferation, yet its molecular description is challenging, because >100 conserved proteins must be spatially and temporally co-ordinated. despite the high importance of a tight co-ordination of cytokinesis with chromosome and organelle segregation, the mechanism for determining the cell division plane is one of the least conserved aspects of cytokinesis in eukaryotic cells. budding and fission yeast have developed fundamentally distinct mechanisms to ensure pr ...201021091496
mips: curated databases and comprehensive secondary data resources in 2010.the munich information center for protein sequences (mips at the helmholtz center for environmental health, neuherberg, germany) has many years of experience in providing annotated collections of biological data. selected data sets of high relevance, such as model genomes, are subjected to careful manual curation, while the bulk of high-throughput data is annotated by automatic means. high-quality reference resources developed in the past and still actively maintained include saccharomyces cerev ...201021109531
survey and analysis of simple sequence repeats in the laccaria bicolor genome, with development of microsatellite is becoming clear that simple sequence repeats (ssrs) play a significant role in fungal genome organization, and they are a large source of genetic markers for population genetics and meiotic maps. we identified ssrs in the laccaria bicolor genome by in silico survey and analyzed their distribution in the different genomic regions. we also compared the abundance and distribution of ssrs in l. bicolor with those of the following fungal genomes: phanerochaete chrysosporium, coprinopsis cinerea, ...201021132299
scanning electron microscopy of mitotic nuclei and chromosomes in filamentous fungi.a new method for scanning electron microscopy (sem) of fungal mitotic nuclei and chromosomes was established for two ascomycetes, cochliobolus heterostrophus and neurospora crassa. nuclei and chromosomes discharged from germling cells by the germ-tube burst method were spread on a surface of a glass slide. the spreads were impregnated with osmium-thiocarbohydrazide for conductive staining, followed by coating with platinum, and observed by field-emission sem. ultrastructure of fungal chromosomes ...200421148846
mapping woronin body position in aspergillus nidulans.the positions of all woronin bodies in five germlings of aspergillus nidulans prepared by plunge freezing and freeze substitution were determined by transmission electron microscopy. as expected, woronin bodies were found near septa. high numbers of morphologically identical organelles were also found in apical regions. to verify that these organelles were authentic woronin bodies, we used antibodies raised against the neurospora crassa woronin body matrix protein hex1. anti-hex1 antibodies labe ...200221156495
characterization of bc-hch, the botrytis cinerea homolog of the neurospora crassahet-c vegetative incompatibility locus, and its use as a population marker.the botrytis cinerea homolog (bc-hch) of nc-het-c and pa-hch (vegetative incompatibility loci of neurospora crassa and podospora anserina respectively) was cloned and sequenced. the gene structure of bc-hch is very close to those of nc-het-c and pa-hch. a pcr-rflp approach on a 1171 bp fragment was used to screen polymorphism at this locus among 117 natural isolates of b. cinerea. restriction patterns by the restriction enzyme hhai fell into two allelic types. moreover, haplotypes at the bc-hch ...200321156611
involvement of a putative response regulator brrg-1 in the regulation of sporulation, sensitivity to fungicides, and osmotic stress in botrytis cinerea.the response regulator protein is a core element of two-component signaling pathway. in this study, we investigated functions of brrg-1 of botrytis cinerea, a gene that encodes a putative response regulator protein, which is homologous to rrg-1 in neurospora crassa. the brrg-1 gene deletion mutant δbrrg1-62 was unable to produce conidia. the mutant showed increased sensitivity to osmotic stress mediated by nacl and kcl, and to oxidative stress generated by h(2)o(2). additionally, the mutant was ...201021161211
progressive cavitating leukoencephalopathy associated with respiratory chain complex i deficiency and a novel mutation in ndufs1.we present clinical, neuroimaging, and molecular data on the identification of a new homozygous c.1783a>g (p.thr595ala) mutation in ndufs1 in two inbred siblings with isolated complex i deficiency associated to a progressive cavitating leukoencephalopathy, a clinical and neuroradiological entity originally related to unknown defects of the mitochondrial energy metabolism. in both sibs, the muscle biopsy showed severe reduction of complex i enzyme activity, which was not obvious in fibroblasts. w ...201121203893
catalytic properties and classification of cellobiose dehydrogenases from ascomycetes.putative cellobiose dehydrogenase (cdh) genes are frequently discovered in various fungi by genome sequencing projects. the expression of cdh, an extracellular flavocytochrome, is well studied in white rot basidiomycetes and is attributed to extracellular lignocellulose degradation. cdh has also been reported for plant-pathogenic or saprotrophic ascomycetes, but the molecular and catalytic properties of these enzymes are currently less investigated. this study links various ascomycetous cdh gene ...201121216904
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
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
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
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
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
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
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
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
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
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
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
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
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
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
a new diet for yeast to improve biofuel 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
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
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
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
[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
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
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
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
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
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
circadian timekeeping in neurospora crassa and synechococcus 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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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