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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
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
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
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
survey and analysis of simple sequence repeats in the laccaria bicolor genome, with development of microsatellite markers.it 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
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
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
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
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
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
aspergillus oryzae type iii polyketide synthase csya is involved in the biosynthesis of 3,5-dihydroxybenzoic acid.as 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
camp signaling pathway controls glycogen metabolism in neurospora crassa by regulating the glycogen synthase gene expression and phosphorylation.the camp-pka signaling pathway plays an important role in many biological processes including glycogen metabolism. in this work we investigated its role in the neurospora crassa glycogen metabolism control using mutant strains affected in components of the pathway, the cr-1 strain deficient in adenylyl cyclase activity therefore has the pka pathway not active, and the mcb strain a temperature-sensitive mutant defective in the regulatory subunit of pka therefore is a strain with constitutively ac ...201019883780
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
genetic and molecular characterization of a cryptochrome from the filamentous fungus neurospora crassa.in 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
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
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
[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
the ndr kinase dbf-2 is involved in regulation of mitosis, conidial development, and glycogen metabolism in neurospora crassa.neurospora crassa dbf-2 encodes an ndr (nuclear dbf2-related) protein kinase, homologous to lats1, a core component of the hippo pathway. this pathway plays important roles in restraining cell proliferation and promoting apoptosis in differentiating cells. here, we demonstrate that dbf-2 is involved in three fundamental processes in a filamentous fungus: cell cycle regulation, glycogen biosynthesis, and conidiation. dbf-2 is predominantly localized to the nucleus, and most (approximately 60%) db ...201019966031
characterization of ctr family genes and the elucidation of their role in the life cycle of neurospora crassa.transcriptional analysis using qrt-pcr of 62 metal ion transporters during conidial germination of neurospora crassa showed a significant up regulation of a hypothetical copper transporter gene, tcu-1, that belongs to the ctr family. herein we characterised the ctr family genes (tcu-1, tcu-2 and tcu-3) and deciphered their role in various developmental phases of the n. crassa life cycle. cross complementation assays in copper uptake mutant of saccharomyces cerevisiae revealed that tcu-1, tcu-2 a ...201020034585
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
aoso protein accumulates at the septal pore in response to various stresses in the filamentous fungus aspergillus oryzae.filamentous ascomycetes form hyphal networks that are compartmentalized by septa which have a perforated pore allowing the passage of cytoplasm and organelles between adjacent hyphal compartments. thus, the septal pore may play an important role in the organized growth of multicellular organisms. upon hyphal injury, the septal pore is plugged by a wound-healing organelle, known as the woronin body, to prevent excessive cytoplasmic leakage. however, the movement of proteins towards the septal por ...201019945422
purification and characterization of a low molecular weight endo-xylanase from mushroom termitomyces clypeatus.a low molecular weight endo-xylanase (ec 3.2.1.8) was purified from an edible mushroom termitomyces clypeatus grown in submerged medium with oat spelt xylan. xylanase was purified to apparent homogeneity by ammonium sulfate fractionation and gel filtration chromatography. its molecular weight was determined by gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 12 kda. the enzyme was found to be most active at 50 degrees c and ph 5.0, being most stab ...201019728166
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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 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
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
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
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
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
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
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 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
fungal photobiology: a synopsis.fungi respond and adapt to many environmental signals including light. the photobiology of fungi has been extensively investigated, but in recent years the identification of the first fungal photoreceptor, wc-1 in the ascomycete neurospora crassa, and the discovery that similar photoreceptors are required for photoreception in other ascomycete, basidiomycete and zygomycete fungi has allowed the molecular characterization of light reception and the early steps of signal transduction in a number o ...201122679584
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
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
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
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
characterization of cytotoxic compound from mangrove derived fungi irpex hydnoides vb4.to investigate the cytotoxic activity of endophytic fungi isolated from mangrove fungi.201123569763
[vegetable cytology. intracellular localization of transaminase alanine-glyoxylate and malate synthetase in neurospora crassa]. 20114979219
a potassium-proton symport in neurospora crassa.combined ion flux and electrophysiological measurements have been used to characterized active transport of potassium by cells of neurospora crassa that have been moderately starved of k+ and then maintained in the presence of millimolar free calcium ions. these conditions elicit a high-affinity (k1/2 = 1-10 microm) potassium uptake system that is strongly depolarizing. current-voltage measurements have demonstrated a k+-associated inward current exceeding (at saturation) half the total current ...20113014042
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
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
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
self/nonself recognition in tuber melanosporum is not mediated by a heterokaryon incompatibility system.vegetative incompatibility is a widespread phenomenon in filamentous ascomycetes, which limits formation of viable heterokaryons. whether this phenomenon plays a role in maintaining the homokaryotic state of the hyphae during the vegetative growth of tuber spp. gene expression, polymorphism analysis as well as targeted in vitro experiments allowed us to test whether a heterokaryon incompatibility (hi) system operates in tuber melanosporum. hi is controlled by different genetic systems, often inv ...201122289772
the kinetics of changes in the fatty acid composition of neurospora crassa lipids after a temperature increase.the fatty acid composition of the total lipids, phospholipids and neutral lipids of log-phase shaker cultures of the bd (band) strain of neurospora crassa, were measured every 2 h for an 8-h period following a temperature increase from 22 to 40 degrees c. for purposes of comparison, the fatty acid composition was also measured when cultures were grown from inoculation at temperatures between 22 and 40 degrees c. in the phospholipids, the temperature jump produced, over a 4-6 hour span, a linear ...20113159434
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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