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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
molecular mechanism of the neurospora circadian oscillator.circadian clocks are the internal time-keeping mechanisms for organisms to synchronize their cellular and physiological processes to the daily light/dark cycles. the molecular mechanisms underlying circadian clocks are remarkably similar in eukaryotes. neurospora crassa, a filamentous fungus, is one of the best understood model organisms for circadian research. in recent years, accumulating data have revealed complex regulation in the neurospora circadian clock at transcriptional, posttranscript ...201021203945
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 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
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
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
role of individual subunits of the neurospora crassa csn complex in regulation of deneddylation and stability of cullin proteins.the cop9 signalosome (csn) is an evolutionarily conserved multifunctional complex that controls ubiquitin-dependent protein degradation in eukaryotes. we found seven csn subunits in neurospora crassa in a previous study, but only one subunit, csn-2, was functionally characterized. in this study, we created knockout mutants for the remaining individual csn subunits in n. crassa. by phenotypic observation, we found that loss of csn-1, csn-2, csn-4, csn-5, csn-6, or csn-7 resulted in severe defects ...201021151958
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
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
evolution and diversity of a fungal self/nonself recognition locus.self/nonself discrimination is an essential feature for pathogen recognition and graft rejection and is a ubiquitous phenomenon in many organisms. filamentous fungi, such as neurospora crassa, provide a model for analyses of population genetics/evolution of self/nonself recognition loci due to their haploid nature, small genomes and excellent genetic/genomic resources. in n. crassa, nonself discrimination during vegetative growth is determined by 11 heterokaryon incompatibility (het) loci. cell ...201021124910
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
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
conidiation of neurospora crassa. 196621090502
biosynthesis of silver, gold and bimetallic nanoparticles using the filamentous fungus neurospora crassa.the development of production processes that can reduce the environmental impact, offer waste reduction and increase energy efficiency is an important step in the field of application of nanotechnology. in this work the filamentous fungus neurospora crassa was screened and found to be successful for the production of mono and bimetallic au/ag nanoparticles (nps). analysis by scanning electron microscopy (sem), energy dispersive x-ray spectroscopy (eds), and transmission electron microscopy (tem) ...201121087843
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
dna methylation and normal chromosome behavior in neurospora depend on five components of a histone methyltransferase complex, dcdc.methylation of dna and of lysine 9 on histone h3 (h3k9) is associated with gene silencing in many animals, plants, and fungi. in neurospora crassa, methylation of h3k9 by dim-5 directs cytosine methylation by recruiting a complex containing heterochromatin protein-1 (hp1) and the dim-2 dna methyltransferase. we report genetic, proteomic, and biochemical investigations into how dim-5 is controlled. these studies revealed dcdc, a previously unknown protein complex including dim-5, dim-7, dim-9, cu ...201021079689
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
activation and localization of protein kinase c in neurospora crassa.the neurosporacrassa protein kinase c (npkc) is reported to be a regulator of light responsive genes. it phosphorylates the light receptor wc-1 and regulates the levels of the circadian clock protein frq and transcription of the light-induced albino-2 gene. in mammals, the conventional and novel isoforms of pkc are activated by diacylglycerol (dag), which induces pkc translocation from the cytoplasm to membranes. to investigate the interaction of npkc and dag in neurospora, we constructed a stra ...201121070858
the de novo cytosine methyltransferase drm2 requires intact uba domains and a catalytically mutated paralog drm3 during rna-directed dna methylation in arabidopsis thaliana.eukaryotic dna cytosine methylation can be used to transcriptionally silence repetitive sequences, including transposons and retroviruses. this silencing is stable between cell generations as cytosine methylation is maintained epigenetically through dna replication. the arabidopsis thaliana dnmt3 cytosine methyltransferase ortholog domains rearranged methyltransferase2 (drm2) is required for establishment of small interfering rna (sirna) directed dna methylation. in mammals piwi proteins and pir ...201021060858
of switches and hourglasses: regulation of subcellular traffic in circadian clocks by phosphorylation.investigation of the phosphorylation of circadian clock proteins has shown that this modification contributes to circadian timing in all model organisms. phosphorylation alters the stability, transcriptional activity and subcellular localization of clock proteins during the course of a day, such that time-of-day-specific phosphorylation encodes information for measuring time and is crucial for the establishment of an approximately 24-h period. one main feature of molecular timekeeping is the day ...201021052092
adenine-requiring mutants of neurospora crassa. 194620999484
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
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
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
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
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
degradation of carbohydrate moieties of arabinogalactan-proteins by glycoside hydrolases from neurospora crassa.arabinogalactan-proteins (agps) are a family of plant proteoglycans having large carbohydrate moieties attached to core-proteins. the carbohydrate moieties of agps commonly have β-(1→3)(1→6)-galactan as the backbone, to which other auxiliary sugars such as l-ara and glca are attached. for the present study, an α-l-arabinofuranosidase belonging to glycoside hydrolase family (ghf) 54, ncaraf1, and an endo-β-(1→6)-galactanase of ghf 5, nc6gal, were identified in neurospora crassa. recombinant ncara ...201020932514
dcaf26, an adaptor protein of cul4-based e3, is essential for dna methylation in neurospora crassa.dna methylation is involved in gene silencing and genome stability in organisms from fungi to mammals. genetic studies in neurospora crassa previously showed that the cul4-ddb1 e3 ubiquitin ligase regulates dna methylation via histone h3k9 trimethylation. however, the substrate-specific adaptors of this ligase that are involved in the process were not known. here, we show that, among the 16 ddb1- and cul4-associated factors (dcafs) encoded in the n. crassa genome, three interacted strongly with ...201020885793
sequence requirements for ribosome stalling by the arginine attenuator peptide.the 5' regions of eukaryotic mrnas often contain upstream open reading frames (uorfs). the neurospora crassa arg-2 uorf encodes the 24-residue arginine attenuator peptide (aap). this regulatory uorf-encoded peptide, which is evolutionarily conserved in fungal transcripts specifying an arginine biosynthetic enzyme, functions as a nascent peptide within the ribosomal tunnel and negatively regulates gene expression. the nascent aap causes ribosomes to stall at the uorf stop codon in response to arg ...201020884617
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
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
modeling a circadian surface.experiments that systematically varied t, τ, and photoperiod in neurospora crassa revealed that the traditional nonparametric and parametric approaches could not explain entrainment for all of the tested conditions. the authors have developed a new approach to understanding entrainment that incorporates several features of the old paradigms but allows exploration of the underlying mechanisms in synchronized clocks, making extrapolations from constant conditions to entrained state unnecessary. it ...201020876814
a circadian surface of entrainment: varying t, τ, and photoperiod in neurospora crassa.the two major prerequisites for a functional circadian system are the generation of an internal day (circadian cycle) and adjusting its length-and phase-to that of the external day (zeitgeber cycle). the generation of circadian cycles can be observed in constant conditions where organisms show a self-sustained, free-running rhythm. their expression depends on the nature of the constant conditions (e.g., constant darkness, dd, or constant light, ll). the mechanism that synchronizes the circadian ...201020876812
temporal and spatial regulation of gene expression during asexual development of neurospora crassa.in this study we profiled spatial and temporal transcriptional changes during asexual sporulation in the filamentous fungus neurospora crassa. aerial tissue was separated from the mycelium to allow detection of genes specific to each tissue. we identified 2641 genes that were differentially expressed during development, which represents ∼25% of the predicted genes in the genome of this model fungus. on the basis of the distribution of functional annotations of 1102 of these genes, we identified ...201020876563
h2b- and h3-specific histone deacetylases are required for dna methylation in neurospora crassa.neurospora crassa utilizes dna methylation to inhibit transcription of heterochromatin. dna methylation is controlled by the histone methyltransferase dim-5, which trimethylates histone h3 lysine 9, leading to recruitment of the dna methyltransferase dim-2. previous work demonstrated that the histone deacetylase (hdac) inhibitor trichostatin a caused a reduction in dna methylation, suggesting involvement of histone deacetylation in dna methylation. we therefore created mutants of each of the fou ...201020876559
α-1,6-mannosylation of n-linked oligosaccharide present on cell wall proteins is required for their incorporation into the cell wall in the filamentous fungus neurospora crassa.the enzyme α-1,6-mannosyltransferase (och-1) is required for the synthesis of galactomannans attached to the n-linked oligosaccharides of neurospora crassa cell wall proteins. the neurospora crassa och-1 mutant has a tight colonial phenotype and a defective cell wall. a carbohydrate analysis of the och-1 mutant cell wall revealed a 10-fold reduction in the levels of mannose and galactose and a total lack of 1,6-linked mannose residues. analysis of the integral cell wall protein from wild-type an ...201020870880
circadian oscillators in eukaryotes.the biological clock, present in nearly all eukaryotes, has evolved such that organisms can adapt to our planet's rotation in order to anticipate the coming day or night as well as unfavorable seasons. as all modern high-precision chronometers, the biological clock uses oscillation as a timekeeping element. in this review, we describe briefly the discovery, historical development, and general properties of circadian oscillators. the issue of temperature compensation (tc) is discussed, and our pr ...201020836046
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
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
vivid interacts with the white collar complex and frequency-interacting rna helicase to alter light and clock responses in neurospora.the photoreceptor and pas/lov protein vivid (vvd) modulates blue-light signaling and influences light and temperature responses of the circadian clock in neurospora crassa. one of the main actions of vvd on the circadian clock is to influence circadian clock phase by regulating levels of the transcripts encoded by the central clock gene frequency (frq). how this regulation is achieved is unknown. here we show that vvd interacts with complexes central for circadian clock and blue-light signaling, ...201020807745
new insights into the circadian clock in chlamydomonas.the unicellular green alga chlamydomonas reinhardtii has long been used in research on circadian rhythm. various circadian rhythms in behavior and physiology including phototaxis, chemotaxis, the cell division cycle, uv sensitivity, stickiness to glass (changes in properties of the cell surface), and starch content are observed in this alga. soon after the isolation of clock mutants in drosophila melanogaster and neurospora crassa, clock mutants were also isolated in chlamydomonas. whereas genes ...201020797685
entrainment elicits period aftereffects in neurospora crassa.circadian clocks continue to oscillate in constant conditions with their own period (tau) and entrain to a cyclic environment by adjusting their intrinsic period to that of the zeitgeber. when circadian clocks are released from entrained to constant conditions, the tau of their initial free-run often depends on the nature of the prior zeitgeber. these postentrainment effects on period (tau-aftereffects) have predominantly been reported for animals but, so far, not fungi. the authors therefore in ...201020795879
physical interaction between vivid and white collar complex regulates photoadaptation in neurospora.photoadaptation, the ability to attenuate a light response on prolonged light exposure while remaining sensitive to escalating changes in light intensity, is essential for organisms to decipher time information appropriately, yet the underlying molecular mechanisms are poorly understood. in neurospora crassa, vivid (vvd), a small lov domain containing blue-light photoreceptor protein, affects photoadaptation for most if not all light-responsive genes. we report that there is a physical interacti ...201020733070
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
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
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
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
a proteomic approach towards the identification of the matrix protein content of the two types of microbodies in neurospora crassa.microbodies (peroxisomes) comprise a class of organelles with a similar biogenesis but remarkable biochemical heterogeneity. here, we purified the two distinct microbody family members of filamentous fungi, glyoxysomes and woronin bodies, from neurospora crassa and analyzed their protein content by hplc/esi-ms/ms. in the purified woronin bodies, we unambiguously identified only hexagonal 1 (hex1), suggesting that the matrix is probably exclusively filled with the hex1 hexagonal crystal. the prot ...201020707002
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
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
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
transcription factors in light and circadian clock signaling networks revealed by genomewide mapping of direct targets for neurospora white collar complex.light signaling pathways and circadian clocks are inextricably linked and have profound effects on behavior in most organisms. here, we used chromatin immunoprecipitation (chip) sequencing to uncover direct targets of the neurospora crassa circadian regulator white collar complex (wcc). the wcc is a blue-light receptor and the key transcription factor of the circadian oscillator. it controls a transcriptional network that regulates ∼20% of all genes, generating daily rhythms and responses to lig ...201020675579
cr(vi) removal on fungal biomass of neurospora crassa: the importance of dissolved organic carbons derived from the biomass to cr(vi) reduction.interactions of toxic cr(vi) with renewable biomaterials are considered an important pathway for cr(vi) removal in ecosystems. biomaterials are susceptible to dissolution, and their dissolved derivatives may provide an alternative to surface-involved pathway for scavenging of cr(vi). in this study, dissolved organic carbon (doc) derived from neurospora crassa biomass was investigated. the proportion of cr(vi) reduction by doc to that on biomass was determined to evaluate the importance of doc to ...201020669959
structure and engineering of l-arabinitol 4-dehydrogenase from neurospora crassa.l-arabinitol 4-dehydrogenase (lad) catalyzes the conversion of l-arabinitol into l-xylulose with concomitant nad(+) reduction. it is an essential enzyme in the development of recombinant organisms that convert l-arabinose into fuels and chemicals using the fungal l-arabinose catabolic pathway. here we report the crystal structure of lad from the filamentous fungus neurospora crassa at 2.6 a resolution. in addition, we created a number of site-directed variants of n. crassa lad that are capable o ...201020655316
in vitro activities of the multifunctional rna silencing polymerase qde-1 of neurospora crassa.qde-1 is an rna- and dna-dependent rna polymerase that has functions in the rna silencing and dna repair pathways of the filamentous fungus neurospora crassa. the crystal structure of the dimeric enzyme has been solved, and the fold of its catalytic core is related closely to that of eukaryotic dna-dependent rna polymerases. however, the specific activities of this multifunctional enzyme are still largely unknown. in this study, we characterized the in vitro activities of the n-terminally trunca ...201020647305
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
neurospora illuminates fungal photoreception.light not only is indispensable as an energy source for life on earth but also serves as an essential environmental cue conveying the information of daily and seasonal time to organisms across different kingdoms. although the molecular mechanisms underlying light responses are actively explored in various light-sensitive organisms, these studies are either hindered by the complexity of the systems or an incomplete familiarity with the light signaling components involved in the scheme. therefore, ...201020637887
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
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
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
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 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
qip, a component of the vegetative rna silencing pathway, is essential for meiosis and suppresses meiotic silencing in neurospora crassa.among the processes that play essential roles in both genome defense and organism survival are those involved in chromosome comparison. they are acutely active in the meiotic cells of neurospora crassa, where they evaluate the mutual identity of homologs by a process we call trans-sensing. when nonsymmetrical regions are found, they are silenced. the known molecular components of this meiotic silencing machinery are related to rna-dependent rna polymerases, argonautes and dicers, suggesting that ...201020592262
real-time fluorescence lifetime imaging system with a 32 x 32 0.13microm cmos low dark-count single-photon avalanche diode array.a compact real-time fluorescence lifetime imaging microscopy (flim) system based on an array of low dark count 0.13microm cmos single-photon avalanche diodes (spads) is demonstrated. fast background-insensitive fluorescence lifetime determination is achieved by use of a recently proposed algorithm called 'integration for extraction method' (iem) [j. opt. soc. am. a 25, 1190 (2008)]. here, iem is modified for a wider resolvability range and implemented on the fpga of the new spad array imager. we ...201020588879
[comparative analysis of respiratory activity in the wild type strain of neurospora crassa and its photoreceptor complex mutants].cell respiratory activity of protoplasts obtained from the wild type of neurospora crassa and photoreceptor complex wcc--white collar 1 (wc-1) and white collar 2 (wc-2)--mutants of neurospora crassa strains was investigated. respiration inhibition by kcn in the presence of 25 mm succinate was similar in all strains and did not exceed 83-85% against control. the significant induction of kcn-resistant respiratory pathway occurred under 1% glucose oxidation in wc-1 and wc-2 mutants if compared with ...201320586289
[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
robustness from flexibility in the fungal circadian clock.robustness is a central property of living systems, enabling function to be maintained against environmental perturbations. a key challenge is to identify the structures in biological circuits that confer system-level properties such as robustness. circadian clocks allow organisms to adapt to the predictable changes of the 24-hour day/night cycle by generating endogenous rhythms that can be entrained to the external cycle. in all organisms, the clock circuits typically comprise multiple interloc ...201020576110
use of an isocitrate lyase promoter-gfp fusion to monitor carbon metabolism of the plant pathogen tapesia yallundae during infection of wheat.abstract green fluorescent protein (gfp) has been used as a vital marker in a variety of species. here, we present the use of a gfp-promoter fusion to visualize carbon metabolism in a pathogenic fungus during growth on defined medium and during infection of plants. isocitrate lyase (icl), a key enzyme in carbon metabolism, is tightly regulated at the transcriptional level, with high levels of expression during 2-carbon growth and no expression during growth on glucose. a gfp-icl promoter fusion ...200020572972
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
atm and atr homologes of neurospora crassa are essential for normal cell growth and maintenance of chromosome integrity.genome integrity is maintained by many cellular mechanisms in eukaryotes. one such mechanism functions during the cell cycle and is known as the dna damage checkpoint. in the filamentous fungus neurospora crassa, mus-9 and mus-21 are homologes of two key factors of the mammalian dna damage checkpoint, atr and atm, respectively. we previously showed that mus-9 and mus-21 mutants are sensitive to dna damage and that each mutant shows a characteristic growth defect: conidia from the mus-9 mutant ha ...201020553930
qip, a protein that converts duplex sirna into single strands, is required for meiotic silencing by unpaired dna.rna interference (rnai) depends on the production of small rna to regulate gene expression in eukaryotes. two rnai systems exist to control repetitive selfish elements in neurospora crassa. quelling targets transgenes during vegetative growth, whereas meiotic silencing by unpaired dna (msud) silences unpaired genes during meiosis. the two mechanisms require common rnai proteins, such as rna-directed rna polymerases, dicers, and argonaute slicers. we have previously demonstrated that, while quell ...201020551436
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
turgor and net ion flux responses to activation of the osmotic map kinase cascade by fludioxonil in the filamentous fungus neurospora crassa.the internal hydrostatic pressure (turgor) of the filamentous fungus neurospora crassa is regulated at about 400-500 kilopascals, primarily by an osmotic map kinase cascade which activates ion uptake from the extracellular medium and glycerol synthesis. in the absence of hyperosmotic stress, the phenylpyrrole fungicide fludioxonil activates the osmotic map kinase cascade, resulting in cell death. turgor, the electrical potential and net ion fluxes were measured after treatment with fludioxonil. ...201020546911
reinforced postmating reproductive isolation barriers in neurospora, an ascomycete microfungus.maladaptive hybridization promotes reinforcement, selection for stringent reproductive isolation barriers during speciation. reinforcement is suspected when barriers between sympatric populations are stronger than allopatric barriers, and particularly when stronger barriers evolve in the species and sex suffering the greatest costs of hybridization. canonically, reinforcement involves premating barriers. selection for postmating barriers is controversial, but theoretically possible. we examined ...201020546092
crystal structure and ligand binding of the mid domain of a eukaryotic argonaute protein.argonaute (ago) proteins are core components of rna-induced silencing complexes and have essential roles in rna-mediated gene silencing. they are characterized by a bilobal architecture, consisting of one lobe containing the amino-terminal and paz domains and another containing the mid and piwi domains. except for the paz domain, structural information on eukaryotic ago domains is not yet available. in this study, we report the crystal structure of the mid domain of the eukaryotic ago protein qd ...201020539312
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
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
deletion of a novel f-box protein, mus-10, in neurospora crassa leads to altered mitochondrial morphology, instability of mtdna and senescence.while mitochondria are renowned for their role in energy production, they also perform several other integral functions within the cell. thus, it is not surprising that mitochondrial dysfunction can negatively impact cell viability. although mitochondria have received an increasing amount of attention in recent years, there is still relatively little information about how proper maintenance of mitochondria and its genomes is achieved. the neurospora crassa mus-10 mutant was first identified thro ...201020516500
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
characterization of the temperature-sensitive mutations un-7 and png-1 in neurospora crassa.the model filamentous fungus neurospora crassa has been studied for over fifty years and many temperature-sensitive mutants have been generated. while most of these have been mapped genetically, many remain anonymous. the mutation in the n. crassa temperature-sensitive lethal mutant un-7 was identified by a complementation based approach as being in the open reading frame designated ncu00651 on linkage group i. other mutations in this gene have been identified that lead to a temperature-sensitiv ...201020502699
catalase evolved to concentrate h2o2 at its active site.catalase is a homo-tetrameric enzyme that has its heme active site deeply buried inside the protein. its only substrate, hydrogen peroxide (h2o2), reaches the heme through a 45 a-long channel. large-subunit catalases, but not small-subunit catalases, have a loop (gate loop) that interrupts the major channel. two accesses lead to a gate that opens the final section of the channel to the heme; gates from the r-related subunits are interconnected. using molecular dynamic simulations of the neurospo ...201020494646
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
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
membrane fluidity determines sensitivity of filamentous fungi to chitosan.the antifungal mode of action of chitosan has been studied for the last 30 years, but is still little understood. we have found that the plasma membrane forms a barrier to chitosan in chitosan-resistant but not chitosan-sensitive fungi. the plasma membranes of chitosan-sensitive fungi were shown to have more polyunsaturated fatty acids than chitosan-resistant fungi, suggesting that their permeabilization by chitosan may be dependent on membrane fluidity. a fatty acid desaturase mutant of neurosp ...201020487294
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
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
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
functional characterization of mat1-1-specific mating-type genes in the homothallic ascomycete sordaria macrospora provides new insights into essential and nonessential sexual regulators.mating-type genes in fungi encode regulators of mating and sexual development. heterothallic ascomycete species require different sets of mating-type genes to control nonself-recognition and mating of compatible partners of different mating types. homothallic (self-fertile) species also carry mating-type genes in their genome that are essential for sexual development. to analyze the molecular basis of homothallism and the role of mating-type genes during fruiting-body development, we deleted eac ...201020435701
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
mapping of lysine methylation and acetylation in core histones of neurospora crassa.core histones are susceptible to a variety of post-translational modifications (ptms), among which methylation and acetylation play critical roles in various chromatin-dependent processes. the nature and biological functions of these ptms have been extensively studied in plants, animals, and yeasts. in contrast, the histone modifications in neurospora crassa, a convenient model organism for multicellular eukaryotes, remained largely undefined. in this study, we used several mass spectrometric te ...201020433192
biotransformation of alpha-cedrol and caryophyllene oxide by the fungus neurospora crassa.incubation of alpha-cedrol and caryophyllene oxide with neurospora crassa afforded 12beta-hydroxy cedrol, 10alpha-hydroxy cedrol, and 3beta-hydroxy cedrol, and 12beta-hydroxy caryophyllene oxide as major metabolites, respectively. the antibacterial and radical scavenging activities of the metabolites were evaluated in vitro using broth microdilution and bioauthographic techniques. however, no significant antibacterial and antioxidant activities were observed when compared with those of standard ...201020433062
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
dna methylation and the formation of heterochromatin in neurospora crassa.studies of the control and function of dna methylation in neurospora crassa have led to a greater understanding of heterochromatin formation. dna methylation in neurospora is dependent on trimethylation of histone h3 lysine 9 (h3k9me3) by the histone methyltransferase, dim-5. the linkage between these two methyl marks is facilitated by heterochromatin protein 1 (hp1), which serves as an adapter protein. hp1 binds to the h3k9me3 and recruits the dna methyltransferase, dim-2. although hp1 links h3 ...201020407471
high efficient gene targeting on the agamous gene in an arabidopsisatlig4 mutant.gene targeting induced by homologous integration of a foreign dna segment into a chromosomal target sequence enables precise disruption or replacement of genes of interest and provides an effective means to analyze gene function, and also becomes an useful technique for breeding. but, integration of introduced dna fragments is predominantly non-homologous in most species. however, we presented high-efficient homologous integration in disruptants of non-homologous end joining (nhej), that is, the ...201020406622
identification of dim-7, a protein required to target the dim-5 h3 methyltransferase to chromatin.functionally distinct chromatin domains are delineated by distinct posttranslational modifications of histones, and in some organisms by differences in dna methylation. proper establishment and maintenance of chromatin domains is critical but not well understood. we previously demonstrated that heterochromatin in the filamentous fungus neurospora crassa is marked by cytosine methylation directed by trimethylated lysine 9 on histone h3 (h3k9me3). h3k9me3 is the product of the dim-5 lysine methylt ...201020404183
de novo assembly of a 40 mb eukaryotic genome from short sequence reads: sordaria macrospora, a model organism for fungal morphogenesis.filamentous fungi are of great importance in ecology, agriculture, medicine, and biotechnology. thus, it is not surprising that genomes for more than 100 filamentous fungi have been sequenced, most of them by sanger sequencing. while next-generation sequencing techniques have revolutionized genome resequencing, e.g. for strain comparisons, genetic mapping, or transcriptome and chip analyses, de novo assembly of eukaryotic genomes still presents significant hurdles, because of their large size an ...201020386741
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
effects of prd circadian clock mutations on frq-less rhythms in neurospora.rhythmic conidiation (spore formation) in neurospora crassa provides a model system for investigating the molecular mechanisms of circadian rhythmicity. a feedback loop involving the frq, wc-1, and wc-2 gene products (frq/ wcc) is an important component of the mechanism; however, rhythmic conidiation can still be observed when these gene products are absent. the nature of the oscillator(s) that drives this frq-less rhythmicity (flo) is an important question in neurospora circadian biology. we ha ...201020348458
marker fusion tagging, a new method for production of chromosomally encoded fusion proteins.a new gene-tagging method (marker fusion tagging [mft]) is demonstrated for neurospora crassa and magnaporthe oryzae. translational fusions between the hygromycin b resistance gene and various markers are inserted into genes of interest by homologous recombination to produce chromosomally encoded fusion proteins. this method can produce tags at any position and create deletion alleles that maintain n- and c-terminal sequences. we show the utility of mft by producing enhanced green fluorescent pr ...201020348390
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
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