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global analyses of ceratocystis cacaofunesta mitochondria: from genome to proteome.the ascomycete fungus ceratocystis cacaofunesta is the causal agent of wilt disease in cacao, which results in significant economic losses in the affected producing areas. despite the economic importance of the ceratocystis complex of species, no genomic data are available for any of its members. given that mitochondria play important roles in fungal virulence and the susceptibility/resistance of fungi to fungicides, we performed the first functional analysis of this organelle in ceratocystis us ...201323394930
both osrecq1 and osrdr1 are required for the production of small rna in response to dna-damage in rice.small rna-mediated gene silencing pathways play important roles in the regulation of development, genome stability and various stress responses in many eukaryotes. recently, a new type of small interfering rnas (qirnas) approximately 20-21 nucleotides long in neurospora crassa have been shown to mediate gene silencing in the dna damage response (ddr) pathway. however, the mechanism for rna silencing in the ddr pathway is largely unknown in plants. here, we report that a class of small rnas (qirn ...201323383126
eukaryotic tpp riboswitch regulation of alternative splicing involving long-distance base pairing.thiamin pyrophosphate (tpp) riboswitches are found in organisms from all three domains of life. examples in bacteria commonly repress gene expression by terminating transcription or by blocking ribosome binding, whereas most eukaryotic tpp riboswitches are predicted to regulate gene expression by modulating rna splicing. given the widespread distribution of eukaryotic tpp riboswitches and the diversity of their locations in precursor messenger rnas (pre-mrnas), we sought to examine the mechanism ...201323376932
glucoamylase is a potential allergen in products expressed in neurospora crassa. 201323352537
transcription of the major neurospora crassa microrna-like small rnas relies on rna polymerase iii.most plant and animal micrornas (mirnas) are transcribed by rna polymerase ii. we previously discovered mirna-like small rnas (milrnas) in the filamentous fungus neurospora crassa and uncovered at least four different pathways for milrna production. to understand the evolutionary origin of milrnas, we determined the roles of polymerases ii and iii (pol ii and pol iii) in milrna transcription. our results show that pol iii is responsible for the transcription of the major milrnas produced in this ...201323349642
using a model filamentous fungus to unravel mechanisms of lignocellulose deconstruction.filamentous fungi are the main source of enzymes used to degrade lignocellulose to fermentable sugars for the production of biofuels. while the most commonly used organism for the production of cellulases in an industrial setting is trichoderma reesei (hypocrea jecorina), recent work in the model filamentous fungus neurospora crassa has shown that the variety of molecular, genetic and biochemical techniques developed for this organism can expedite analyses of the complexities involved in the uti ...201323339486
nonself recognition through intermolecular disulfide bond formation of ribonucleotide reductase in neurospora.type i ribonucleotide reductases (rnrs) are conserved across diverse taxa and are essential for the conversion of rna into dna precursors. in neurospora crassa, the large subunit of rnr (un-24) is unusual in that it also has a nonself recognition function, whereby coexpression of oak ridge (or) and panama (pa) alleles of un-24 in the same cell leads to growth inhibition and cell death. we show that coexpressing these incompatible alleles of un-24 in n. crassa results in a high molecular weight u ...201323335337
biotechnological production of ethanol from renewable resources by neurospora crassa: an alternative to conventional yeast fermentations?microbial production of ethanol might be a potential route to replace oil and chemical feedstocks. bioethanol is by far the most common biofuel in use worldwide. lignocellulosic biomass is the most promising renewable resource for fuel bioethanol production. bioconversion of lignocellulosics to ethanol consists of four major unit operations: pretreatment, hydrolysis, fermentation, and product separation/distillation. conventional bioethanol processes for lignocellulosics apply commercial fungal ...201323318834
reactive oxygen species can modulate circadian phase and period in neurospora crassa.reactive oxygen species (ros) may serve as signals coupling metabolism to other cell functions. in addition to being by-products of normal metabolism, they are generated at elevated levels under environmental stress situations. we analyzed how reactive oxygen species affect the circadian clock in the model organism neurospora crassa. in light/dark cycles, an increase in the levels of reactive oxygen species advanced the phase of both the conidiation rhythm and the expression of the clock gene fr ...201323277144
deletion and expression analysis of beta-(1,3)-glucanosyltransferase genes in neurospora crassa.gpi(glycosylphosphatidylinositol)-anchored beta-(1,3)-glucanosyltransferases play an active role in cell wall biosynthesis in fungi. neurospora crassa has 5 putative beta-(1,3)-glucanosyltransferase genes, namely, gel-1, gel-2, gel-3, gel-4, and gel-5, in its genome. among them, the gel-3 gene is constitutively expressed at the highest level in growing hyphae, whereas gel-1 is expressed at the lowest level. the gel-3 deletion mutant displayed slow growth, while other gel gene disruptants exhibit ...201323274249
[organization of mitochondria in the growing hyphae of neurospora crassa].in vivo fluorescent labeling of mitochondria in neurospora crassa showed the concentration of filamentous mitochondria within 30 μm of apex in growing hyphae. these mitochondrial assemblies propagated forward with the elongation of hyphae, split and segregated as the growing tip bifurcated and formed de novo when new branches formed farther away from the apex. the efficiency of the mitochondria concentration in the apical 30 μm zone is related to the growth rate and identical in hyphae cultivate ...201325509139
the essential phosphoinositide kinase mss-4 is required for polar hyphal morphogenesis, localizing to sites of growth and cell fusion in neurospora crassa.fungal hyphae and plant pollen tubes are among the most highly polarized cells known and pose extraordinary requirements on their cell polarity machinery. cellular morphogenesis is driven through the phospholipid-dependent organization at the apical plasma membrane. we characterized the contribution of phosphoinositides (pis) in hyphal growth of the filamentous ascomycete neurospora crassa. mss-4 is an essential gene and its deletion resulted in spherically growing cells that ultimately lyse. tw ...201223272106
single amino acid substitutions in hxt2.4 from scheffersomyces stipitis lead to improved cellobiose fermentation by engineered saccharomyces cerevisiae.saccharomyces cerevisiae cannot utilize cellobiose, but this yeast can be engineered to ferment cellobiose by introducing both cellodextrin transporter (cdt-1) and intracellular β-glucosidase (gh1-1) genes from neurospora crassa. here, we report that an engineered s. cerevisiae strain expressing the putative hexose transporter gene hxt2.4 from scheffersomyces stipitis and gh1-1 can also ferment cellobiose. this result suggests that hxt2.4p may function as a cellobiose transporter when hxt2.4 is ...201323263959
efficient sequential repetitive gene deletions in neurospora crassa employing a self-excising β-recombinase/six cassette.despite its long-standing history as a model organism, neurospora crassa has limited tools for repetitive gene deletions utilizing recyclable self-excising marker systems. here we describe, for the first time, the functionality of a bacterial recombination system employing β-recombinase acting on six recognition sequences (β-rec/six) in n. crassa, which allowed repetitive site-specific gene deletion and marker recycling. we report generating the mus-51 deletion strain using this system, recyclin ...201323246910
[effect of oxylipins on neurospora crassa growth and differentiation ion]. 201223234069
a uvs-5 strain is deficient for a mitofusin gene homologue, fzo1, involved in maintenance of long life span in neurospora crassa.mitochondria are highly dynamic organelles that continuously fuse and divide. to maintain mitochondria, cells establish an equilibrium of fusion and fission events, which are mediated by dynamin-like gtpases. we previously showed that an mus-10 strain, a mutagen-sensitive strain of the filamentous fungus neurospora crassa, is defective in an f-box protein that is essential for the maintenance of mitochondrial dna (mtdna), long life span, and mitochondrial morphology. similarly, a uvs-5 mutant ac ...201323223037
identification of genes required for alternative oxidase production in the neurospora crassa gene knockout library.the alternative oxidase (aox) of neurospora crassa transfers electrons from ubiquinol to oxygen. the enzyme is not expressed under normal conditions. however, when the function of the standard electron transport chain is compromised, aox is induced, providing cells with a means to continue respiration and growth. induction of the enzyme represents a form of retrograde regulation because aox is encoded by a nuclear gene that responds to signals produced from inefficiently functioning mitochondria ...201223173086
variation in mitochondrial genome primary sequence among whole-genome-sequenced strains of neurospora crassa.eighteen classical mutant strains of neurospora crassa were subject to whole genome sequence analysis and the mitochondrial genome is analyzed. overall, the mitochondrial genomes of the classical mutant strains are 99.45 to 99.98 % identical to the reference genome. two-thirds of the snps and three-fourths of indels identified in this analysis are shared among more than one strain. most of the limited variability in mitochondrial genome sequence is neutral with regard to protein structure. despi ...201223155504
circadian activation of the mitogen-activated protein kinase mak-1 facilitates rhythms in clock-controlled genes in neurospora crassa.the circadian clock regulates the expression of many genes involved in a wide range of biological functions through output pathways such as mitogen-activated protein kinase (mapk) pathways. we demonstrate here that the clock regulates the phosphorylation, and thus activation, of the mapks mak-1 and mak-2 in the filamentous fungus neurospora crassa. in this study, we identified genetic targets of the mak-1 pathway, which is homologous to the cell wall integrity pathway in saccharomyces cerevisiae ...201323125351
the guanine nucleotide exchange factor ric8 regulates conidial germination through gα proteins in neurospora crassa.heterotrimeric g protein signaling is essential for normal hyphal growth in the filamentous fungus neurospora crassa. we have previously demonstrated that the non-receptor guanine nucleotide exchange factor ric8 acts upstream of the gα proteins gna-1 and gna-3 to regulate hyphal extension. here we demonstrate that regulation of hyphal extension results at least in part, from an important role in control of asexual spore (conidia) germination. loss of gna-3 leads to a drastic reduction in conidia ...201223118921
production of four neurospora crassa lytic polysaccharide monooxygenases in pichia pastoris monitored by a fluorimetric assay. 201223102010
the common ancestral core of vertebrate and fungal telomerase rnas.telomerase is a ribonucleoprotein with an intrinsic telomerase rna (ter) component. within yeasts, ter is remarkably large and presents little similarity in secondary structure to vertebrate or ciliate ters. to better understand the evolution of fungal telomerase, we identified 74 ters from pezizomycotina and taphrinomycotina subphyla, sister clades to budding yeasts. we initially identified ter from neurospora crassa using a novel deep-sequencing-based approach, and homologous ter sequences fro ...201323093598
the neurospora crassa chr-1 gene is up-regulated by chromate and its encoded chr-1 protein causes chromate sensitivity and chromium accumulation.the chra membrane protein belongs to the chr superfamily of chromate ion transporters, which includes homologues from bacteria, archaea and eukaryotes. bacterial chra homologues confer chromate resistance by exporting chromate ions from the cell's cytoplasm. the neurospora crassa strain 74-a chr-1 gene encodes a putative chr-1 protein of 507 amino acid residues, which belongs to the chr superfamily. rt-pcr assays showed that expression of the chr-1 gene was up-regulated by chromate exposure of n ...201223085746
light-inducible system for tunable protein expression in neurospora crassa.filamentous fungi are important model systems for understanding eukaryotic cellular processes, including the study of protein expression. a salient feature of fungi is the ability of the protein-processing machinery to perform all of the extensive posttranslational modifications needed in the complex world of eukaryotic organisms, making them great hosts for production of eukaryotic proteins. in the model organism neurospora crassa, several regulatable promoters have been used for heterologous g ...201223050231
blue light acts as a double-edged sword in regulating sexual development of hypocrea jecorina (trichoderma reesei).the industrially important cellulolytic filamentous fungus trichoderma reesei is the anamorph of the pantropical ascomycete hypocrea jecorina. h. jecorina cbs999.97 strain undergoes a heterothallic reproductive cycle, and the mating yields fertilized perithecia imbedded in stromata. asci in the perithecia contain 16 linearly arranged ascospores. here, we investigated h. jecorina sexual development under different light regimes, and found that visible light was dispensable for sexual development ...201223028710
the ndr kinase scaffold hym1/mo25 is essential for mak2 map kinase signaling in neurospora crassa.cell communication is essential for eukaryotic development, but our knowledge of molecules and mechanisms required for intercellular communication is fragmentary. in particular, the connection between signal sensing and regulation of cell polarity is poorly understood. in the filamentous ascomycete neurospora crassa, germinating spores mutually attract each other and subsequently fuse. during these tropic interactions, the two communicating cells rapidly alternate between two different physiolog ...201223028357
the blue light receptor complex wc-1/2 of schizophyllum commune is involved in mushroom formation and protection against phototoxicity.blue light is necessary for initiation of mushroom formation in schizophyllum commune. the genome of this basidiomycete contains homologues of the blue light receptor genes wc-1 and wc-2 of neurospora crassa. here, it is shown that inactivation of either or both of these genes in s. commune results in a blind phenotype. mushroom formation was abolished in dikaryons and they formed symmetrical instead of asymmetrical colonies. development was restored in a temperature dependent way in a δwc-2δwc- ...201322998561
a mouse neurodegenerative dynein heavy chain mutation alters dynein motility and localization in neurospora crassa.cytoplasmic dynein is responsible for the transport and delivery of cargoes in organisms ranging from humans to fungi. dysfunction of dynein motor machinery due to mutations in dynein or its activating complex dynactin can result in one of several neurological diseases in mammals. the mouse legs at odd angles (loa) mutation in the tail domain of the dynein heavy chain has been shown to lead to progressive neurodegeneration in mice. the mechanism by which the loa mutation affects dynein function ...201222991199
the pmr gene, encoding a ca2+-atpase, is required for calcium and manganese homeostasis and normal development of hyphae and conidia in neurospora crassa.the pmr gene is predicted to encode a ca(2+)-atpase in the secretory pathway. we examined two strains of neurospora crassa that lacked pmr: the δpmr strain, in which pmr was completely deleted, and pmr(rip), in which the gene was extensively mutated. both strains had identical, complex phenotypes. compared to the wild type, these strains required high concentrations of calcium or manganese for optimal growth and had highly branched, slow-growing hyphae. they conidiated poorly, and the shape and ...201222983986
improved cellulase production via disruption of pde01641 in cellulolytic fungus penicillium decumbens.to investigate the function of pde01641, the homologue of neurospora crassa ncu05137, during cellulase production in penicillium decumbens, the pde01641 encoding gene was knocked out in the present work. the pde01641 deletion mutant showed enhanced cellobiohydrolase activity and β-glucosidase activity compared to the parental strain 114-2. increased transcription of the main cellulase and hemicellulase genes in δpde01641 gave evidence that pde01641 might affect the process associated with the re ...201222981621
physiological significance of network organization in fungi.the evolution of multicellularity has occurred in diverse lineages and in multiple ways among eukaryotic species. for plants and fungi, multicellular forms are derived from ancestors that failed to separate following cell division, thus retaining cytoplasmic continuity between the daughter cells. in networked organisms, such as filamentous fungi, cytoplasmic continuity facilitates the long-distance transport of resources without the elaboration of a separate vascular system. nutrient translocati ...201222962278
suppression subtractive hybridization and comparative expression analysis to identify developmentally regulated genes in filamentous fungi.ascomycetes differentiate four major morphological types of fruiting bodies (apothecia, perithecia, pseudothecia and cleistothecia) that are derived from an ancestral fruiting body. thus, fruiting body differentiation is most likely controlled by a set of common core genes. one way to identify such genes is to search for genes with evolutionary conserved expression patterns. using suppression subtractive hybridization (ssh), we selected differentially expressed transcripts in pyronema confluens ...201322961396
heterologous gene expression in filamentous fungi.filamentous fungi are critical to production of many commercial enzymes and organic compounds. fungal-based systems have several advantages over bacterial-based systems for protein production because high-level secretion of enzymes is a common trait of their decomposer lifestyle. furthermore, in the large-scale production of recombinant proteins of eukaryotic origin, the filamentous fungi become the vehicle of choice due to critical processes shared in gene expression with other eukaryotic organ ...201222958526
defective valyl-trna synthetase hampers the mitochondrial respiratory chain in neurospora crassa.respiratory chain deficiency can result from alterations in mitochondrial and/or cytosolic protein synthesis due to the dual genetic origin of mitochondrial oxidative phosphorylation. in the present paper we report a point mutation (d750g) in the bifunctional vars (valyl-trna synthetase) of the fungus neurospora crassa, associated with a temperature-sensitive phenotype. analysis of the mutant strain revealed decreased steady-state levels of vars and a clear reduction in the rate of mitochondrial ...201222957697
glycogen synthase kinase is a regulator of the circadian clock of neurospora crassa.timekeeping by circadian clocks relies upon precise adjustment of expression levels of clock proteins. here we identify glycogen synthase kinase (gsk) as a novel and critical component of the circadian clock of neurospora crassa that regulates the abundance of its core transcription factor white collar complex (wcc) on a post-transcriptional level. we show that gsk specifically binds and phosphorylates both subunits of the wcc. reduced expression of gsk promotes an increased accumulation of wc-1 ...201222955278
molecular analysis of mutants of the neurospora adenylosuccinate synthetase locus.the ad-8 gene of neurospora crassa, in addition to being used for the study of purine biology, has been extensively studied as a model for gene structure, mutagenesis and intralocus recombination. because of this there is an extensive collection of well-characterized n. crassa ad-8 mutants in the fungal genetics stock center collection. among these are spontaneous mutants and mutants induced with x-ray, uv or chemical mutagens. the specific lesions in these mutants have been genetically mapped a ...201222942090
evidence for tryptophan being a signal molecule that inhibits conidial anastomosis tube fusion during colony initiation in neurospora crassa.mycelial interconnectedness achieved by hyphal fusion has been hypothesized to facilitate the distribution and sharing of nutrients between different parts of a mycelium, especially when nutrients are heterogeneously distributed in the environment. however, the link between environmental nutrient availability and hyphal fusion is little understood. here, we report that amino acids and extracellular ph regulate conidial anastomosis tube (cat) fusion during colony initiation in neurospora crassa. ...201222939838
the saccharomyces cerevisiae bem1 homologue in neurospora crassa promotes co-ordinated cell behaviour resulting in cell fusion.directed growth or movement is a common feature of microbial development and propagation. in polar growing filamentous fungi, directed growth requires the interaction of signal sensing machineries with factors controlling polarity and cell tip extension. in neurospora crassa an unusual mode of cell-cell signalling mediates mutual attraction of germinating spores, which subsequently fuse. during directed growth of the two fusion partners, the cells co-ordinately alternate between two physiologica ...201222906237
genetic and physical interactions between gα subunits and components of the gβγ dimer of heterotrimeric g proteins in neurospora crassa.heterotrimeric g proteins are critical regulators of growth and asexual and sexual development in the filamentous fungus neurospora crassa. three gα subunits (gna-1, gna-2, and gna-3), one gβ subunit (gnb-1), and one gγ subunit (gng-1) have been functionally characterized, but genetic epistasis relationships between gβ and gα subunit genes have not been determined. physical association between gnb-1 and flag-tagged gng-1 has been previously demonstrated by coimmunoprecipitation, but knowledge of ...201222903975
importance of map kinases during protoperithecial morphogenesis in neurospora crassa.in order to produce multicellular structures filamentous fungi combine various morphogenetic programs that are fundamentally different from those used by plants and animals. the perithecium, the female sexual fruitbody of neurospora crassa, differentiates from the vegetative mycelium in distinct morphological stages, and represents one of the more complex multicellular structures produced by fungi. in this study we defined the stages of protoperithecial morphogenesis in the n. crassa wild type i ...201222900028
wsc-1 and ham-7 are mak-1 map kinase pathway sensors required for cell wall integrity and hyphal fusion in neurospora crassa.a large number of cell wall proteins are encoded in the neurospora crassa genome. strains carrying gene deletions of 65 predicted cell wall proteins were characterized. deletion mutations in two of these genes (wsc-1 and ham-7) have easily identified morphological and inhibitor-based defects. their phenotypic characterization indicates that ham-7 and wsc-1 function during cell-to-cell hyphal fusion and in cell wall integrity maintenance, respectively. wsc-1 encodes a transmembrane protein with e ...201222879952
physical association of the wc-1 photoreceptor and the histone acetyltransferase ngf-1 is required for blue light signal transduction in neurospora crassa.in neurospora crassa and other filamentous fungi, light-dependent-specific phenomena are regulated by transcription factors wc-1 and wc-2. in addition to its transcriptional activity, wc-1 is able to directly sense light stimuli through a lov sensor domain. its location in the nucleus and heterodimerization with wc-2, together with the presence of a zinc-finger dna-binding domain and an environmental sensor domain, all resemble the functional evolutionary architecture adopted by vertebrate nucle ...201222875992
in vivo conformation and replication intermediates of circular mitochondrial plasmids in neurospora and cryphonectria parasitica.the in vivo conformation and replication intermediates of fungal circular mitochondrial plasmids and plasmid-like mitochondrial element (plmes) were analyzed by two-dimensional gel electrophoresis and electron microscopy. plasmids with circular restriction maps exist predominantly as circular molecules and were found to replicate by rolling circle mechanisms. however, the reverse transcriptase-encoding mauriceville plasmid of neurospora crassa was observed to replicate by two possible mechanisms ...201222862920
diversification of a protein kinase cascade: ime-2 is involved in nonself recognition and programmed cell death in neurospora crassa.kinase cascades and the modification of proteins by phosphorylation are major mechanisms for cell signaling and communication, and evolution of these signaling pathways can contribute to new developmental or environmental response pathways. the saccharomyces cerevisiae kinase ime2 has been well characterized for its role in meiosis. however, recent studies have revealed alternative functions for ime2 in both s. cerevisiae and other fungi. in the filamentous fungus neurospora crassa, the ime2 hom ...201222813893
graphite electrodes modified with neurospora crassa cellobiose dehydrogenase: comparative electrochemical characterization under direct and mediated electron transfer.the electrochemical characterization of a class ii cellobiose dehydrogenase (cdh), from the ascomycete fungus neurospora crassa, adsorbed on graphite (g), was performed in regard to direct (det) and mediated electron transfer (met). the effects of the applied potential, mediator (1,4 benzoquinone) concentration and flow carrier ph on the amperometric response of the g/cdh modified electrodes were investigated under flow conditions. from the calibration curves, recorded at two ph values (5.2 and ...201222809780
functional analysis of glycoside hydrolase family 18 and 20 genes in neurospora crassa.glycoside hydrolase family 18 contains hydrolytic enzymes with chitinase or endo-n-acetyl-β-d-glucosaminidase (engase) activity, while glycoside hydrolase family 20 contains enzymes with β-n-acetylhexosaminidase (nagase) activity. chitinases and nagases are involved in chitin degradation. chitinases are phylogenetically divided into three main groups (a, b and c), each further divided into subgroups. in this study, we investigated the functional role of 10 neurospora crassa genes that encode chi ...201222796096
biophysical characterization of the recombinant importin-α from neurospora crassa.neurospora crassa has been widely used as a model organism and contributed to the development of biochemistry and molecular biology by allowing the identification of many metabolic pathways and mechanisms responsible for gene regulation. nuclear proteins are synthesized in the cytoplasm and need to be translocated to the nucleus to exert their functions which the importin-α receptor has a key role for the classical nuclear import pathway. in an attempt to get structural information of the nuclea ...201322789101
the translin-trax complex (c3po) is a ribonuclease in trna processing.conserved translin-trax complexes, also known as c3pos, have been implicated in many biological processes, but how they function remains unclear. recently, c3po was shown to be an endornase that promotes rna interference (rnai) in animal cells. here, we show that c3po does not play a significant role in rnai in the filamentous fungus neurospora crassa. instead, the neurospora c3po functions as an rnase that removes the 5' pre-trna fragments after the processing of pre-trnas by rnase p. in additi ...201222773104
cdr4 is the major contributor to azole resistance among four pdr5p-like abc transporters in neurospora crassa.pdr5p-like abc transporters play a significant role in azole resistance in saccharomyces cerevisiae and candida spp. most of filamentous fungal species have multiple pdr5p homologues. in this study, phylogenic analysis identified that filamentous fungi have at least two phylogenically distant groups of pdr5p homologues. one contains pmr1-like pdr5p homologues while the other contains both atrf-like and atrb-like pdr5p homologues. neurospora crassa has a total of four genes encoding pdr5p homolog ...201222749171
analysis of the sulfur-regulated control of the cystathionine γ-lyase gene of neurospora crassa.cystathionine γ-lyase plays a key role in the transsulfuration pathway through its primary reaction of catalyzing the formation of cysteine from cystathionine. the neurospora crassa cystathionine γ-lyase gene (cys-16(+)) is of particular interest in dissecting the regulation and dynamics of transsulfuration. the aim of this study was to determine the regulatory connection of cys-16(+) to the neurospora sulfur regulatory network. in addition, the cys-16(+) promoter was characterized with the goal ...201222748183
rna interference pathways in fungi: mechanisms and functions.rna interference (rnai) is a conserved eukaryotic gene regulatory mechanism that uses small noncoding rnas to mediate posttranscriptional/transcriptional gene silencing. the fission yeast schizosaccharomyces pombe and the filamentous fungus neurospora crassa have served as important model systems for rnai research. studies on these two organisms and other fungi have contributed significantly to our understanding of the mechanisms and functions of rnai in eukaryotes. in addition, surprisingly div ...201222746336
characterization of the two neurospora crassa cellobiose dehydrogenases and their connection to oxidative cellulose degradation.the genome of neurospora crassa encodes two different cellobiose dehydrogenases (cdhs) with a sequence identity of only 53%. so far, only cdh iia, which is induced during growth on cellulose and features a c-terminal carbohydrate binding module (cbm), was detected in the secretome of n. crassa and preliminarily characterized. cdh iib is not significantly upregulated during growth on cellulosic material and lacks a cbm. since cdh iib could not be identified in the secretome, both cdhs were recomb ...201222729546
neurosporaxanthin production by neurospora and fusarium.the orange pigmentation of the ascomycete fungi neurospora and fusarium is mainly due to the accumulation of neurosporaxanthin, a carboxylic apocarotenoid whose possible biotechnological applications have not been investigated. from the discovery of the first enzyme of the biosynthetic pathway in 1989, the prenyltransferase al-3, to the recent identification of an aldehyde dehydrogenase responsible for the last biosynthetic step, all the enzymes and biochemical reactions needed for neurosporaxan ...201222711132
rho1 and rho2 share partially overlapping functions in the regulation of cell wall integrity and hyphal polarity in neurospora crassa.rho proteins are key regulators of cellular morphogenesis, but their function in filamentous fungi is poorly understood. by generating conditional rho-1 mutants, we dissected the function of the essential gtpase rho1 in cell polarization and maintenance of cell wall integrity in neurospora crassa. we identified ncu00668/rgf1 as rho1-specific exchange factor, which controls actin organization and the cell wall integrity mak1 map kinase pathway through the direct interaction of active rho1 with th ...201222703449
the neurospora crassa dfg5 and dcw1 genes encode α-1,6-mannanases that function in the incorporation of glycoproteins into the cell wall.the covalent cross-linking of cell wall proteins into the cell wall glucan/chitin matrix is an important step in the biogenesis of the fungal cell wall. we demonstrate that the neurospora crassa dfg5 (ncu03770) and dcw1 (ncu08127) enzymes function in vivo to cross-link glycoproteins into the cell wall. mutants lacking dfg5 or dcw1 release slightly elevated levels of cell wall proteins into their growth medium. mutants lacking both dfg5 and dcw1 have substantially reduced levels of cell wall prot ...201222701726
high-resolution spatiotemporal analysis of gene expression in real time: in vivo analysis of circadian rhythms in neurospora crassa using a frequency-luciferase translational reporter.the pacemaker of the neurospora circadian clock is composed of a transcriptional-translational feedback loop that has been intensively studied during the last two decades. invaluable information has been derived from measuring the expression of the central clock component frequency (frq) under liquid culture conditions. direct analyses of frq mrna and protein levels on solid media - where overt circadian rhythms are normally visualized - have not been trivial due to technical issues. nevertheles ...201222695169
coronin is a component of the endocytic collar of hyphae of neurospora crassa and is necessary for normal growth and morphogenesis.coronin plays a major role in the organization and dynamics of actin in yeast. to investigate the role of coronin in a filamentous fungus (neurospora crassa), we examined its subcellular localization using fluorescent proteins and the phenotypic consequences of coronin gene (crn-1) deletion in hyphal morphogenesis, spitzenkörper behavior and endocytosis. coronin-gfp was localized in patches, forming a subapical collar near the hyphal apex; significantly, it was absent from the apex. the subapica ...201222693603
use of fluorescent protein to analyse recombination at three loci in neurospora crassa.we have inserted a histone h1-gfp fusion gene adjacent to three loci on different chromosomes of neurospora crassa and made mating pairs in which a wild type version of gfp is crossed to one with a mutation in the 5' end of gfp. the loci are his-3, am and his-5, chosen because recombination mechanisms appear to differ between his-3 and am, and because crossing over adjacent to his-5, like his-3, is regulated by rec-2. at his-3, the frequencies of crossing over between gfp and the centromere and ...201222691725
analyses of dynein heavy chain mutations reveal complex interactions between dynein motor domains and cellular dynein functions.cytoplasmic dynein transports cargoes for a variety of crucial cellular functions. however, since dynein is essential in most eukaryotic organisms, the in-depth study of the cellular function of dynein via genetic analysis of dynein mutations has not been practical. here, we identify and characterize 34 different dynein heavy chain mutations using a genetic screen of the ascomycete fungus neurospora crassa, in which dynein is nonessential. interestingly, our studies show that these mutations seg ...201222649085
concentration-dependent mechanisms of cell penetration and killing by the de novo designed antifungal hexapeptide paf26.recent evidence indicates that antimicrobial peptides can kill microbes in more complex ways than just by membrane permeabilization. in this study, the mechanism of internalization of the de novo designed cationic hexapeptide paf26 has been characterized in detail using neurospora crassa. live-cell imaging of fluorescently labelled paf26, organelle probes and mutants indicate that the peptide is endocytically internalized at low fungicidal concentrations (2.0-5 µm). at these concentrations, paf2 ...201222646057
role of fig1, a component of the low-affinity calcium uptake system, in growth and sexual development of filamentous fungi.the function of fig1, a transmembrane protein of the low-affinity calcium uptake system (lacs) in fungi, was examined for its role in the growth and development of the plant pathogen fusarium graminearum. the δfig1 mutants failed to produce mature perithecia, and sexual development was halted prior to the formation of perithecium initials. the loss of fig1 function also resulted in a reduced vegetative growth rate. macroconidium production was reduced 70-fold in the δfig1 mutants compared to the ...201222635922
sex-specific gene expression during asexual development of neurospora crassa.the impact of loci that determine sexual identity upon the asexual, dominant stage of fungal life history has been well studied. to investigate their impact, expression differences between strains of different mating type during asexual development were assayed, with rna sampled from otherwise largely isogenic mat a and mat a strains of neurospora crassa at early, middle, and late clonal stages of development. we observed significant differences in overall gene expression between mating types ac ...201222626843
the functions of grainy head-like proteins in animals and fungi and the evolution of apical extracellular barriers.the grainy head (grh) family of transcription factors are crucial for the development and repair of epidermal barriers in all animals in which they have been studied. this is a high-level functional conservation, as the known structural and enzymatic genes regulated by grh proteins differ between species depending on the type of epidermal barrier being formed. interestingly, members of the cp2 superfamily of transcription factors, which encompasses the grh and lsf families in animals, are also f ...201222590528
neurospora cop9 signalosome integrity plays major roles for hyphal growth, conidial development, and circadian function.the cop9 signalosome (csn) is a highly conserved multifunctional complex that has two major biochemical roles: cleaving nedd8 from cullin proteins and maintaining the stability of crl components. we used mutation analysis to confirm that the jamm domain of the csn-5 subunit is responsible for nedd8 cleavage from cullin proteins in neurospora crassa. point mutations of key residues in the metal-binding motif (ex(n)hxhx(10)d) of the csn-5 jamm domain disrupted csn deneddylation activity without in ...201222589747
on-target labeling of intracellular metabolites combined with chemical mapping of individual hyphae revealing cytoplasmic relocation of isotopologues.a microscale analytical platform integrating microbial cell culture, isotopic labeling, along with visual and mass spectrometric imaging with single-cell resolution has been developed and applied in the monitoring of cellular metabolism in fungal mycelium. the method implements open chips with a two-dimensional surface pattern composed of hydrophobic and hydrophilic zones. two hydrophilic islands are used as medium reservoirs, while the hydrophobic area constitutes the support for the growing ae ...201222583035
production of platinum nanoparticles and nanoaggregates using neurospora crassa.fungal biomass and fungal extract of the nonpathogenic fungus neurospora crassa were successfully used as reducing agents for the biosynthesis of platinum nanoparticles (ptnps). the experiment was carried out by exposing the fungal biomass or the fungal extract to a 0.001 m precursor solution of hexachloroplatinic(iv) acid (h2ptcl6). a change of color of the biomass from pale yellow to dark brown was the first indication of possible formation of ptnps by the fungus. subsequent analyses confirmed ...201222580320
structural basis for substrate targeting and catalysis by fungal polysaccharide monooxygenases.the use of cellulases remains a major cost in the production of renewable fuels and chemicals from lignocellulosic biomass. fungi secrete copper-dependent polysaccharide monooxygenases (pmos) that oxidatively cleave crystalline cellulose and improve the effectiveness of cellulases. however, the means by which pmos recognize and cleave their substrates in the plant cell wall remain unclear. here, we present structures of neurospora crassa pmo-2 and pmo-3 at 1.10 and 1.37 å resolution, respectivel ...201222578542
bioconversion of dilute-acid pretreated sorghum bagasse to ethanol by neurospora crassa.bioethanol production from sweet sorghum bagasse (sb), the lignocellulosic solid residue obtained after extraction of sugars from sorghum stalks, can further improve the energy yield of the crop. the aim of the present work was to evaluate a cost-efficient bioconversion of sb to ethanol at high solids loadings (16 % at pretreatment and 8 % at fermentation), low cellulase activities (1-7 fpu/g sb) and co-fermentation of hexoses and pentoses. the fungus neurospora crassa dsm 1129 was used, which e ...201222573272
yeast adaptor protein, nbp2p, is conserved regulator of fungal ptc1p phosphatases and is involved in multiple signaling pathways.nbp2p is an src homology 3 (sh3) domain-containing yeast protein that is involved in a variety of cellular processes. this small adaptor protein binds to a number of different proteins through its sh3 domain, and a region n-terminal to the sh3 domain binds to the protein phosphatase, ptc1p. despite its involvement in a large number of physical and genetic interactions, the only well characterized function of nbp2p is to recruit ptc1p to the high osmolarity glycerol pathway, which results in down ...201222570491
enforcing silencing: dynamic hp1 complexes in neurospora.analysis of the neurospora crassa chromodomain protein cdp-2, a component of a newly characterized hp1-containing complex, reveals a second gene-silencing mechanism and provides insights into the dynamic nature of chromatin domains that possess shared components.201222551706
the arginine attenuator peptide interferes with the ribosome peptidyl transferase center.the fungal arginine attenuator peptide (aap) is encoded by a regulatory upstream open reading frame (uorf). the aap acts as a nascent peptide within the ribosome tunnel to stall translation in response to arginine (arg). the effect of aap and arg on ribosome peptidyl transferase center (ptc) function was analyzed in neurospora crassa and wheat germ translation extracts using the transfer of nascent aap to puromycin as an assay. in the presence of a high concentration of arg, the wild-type aap in ...201222508989
heterochromatin protein 1 forms distinct complexes to direct histone deacetylation and dna methylation.dna methylation, methylation of histone h3 at lys9 (h3k9me3) and hypoacetylated histones are common molecular features of heterochromatin. important details of their functions and inter-relationships remain unclear, however. in neurospora crassa, h3k9me3 directs dna methylation through a complex containing heterochromatin protein 1 (hp1) and the dna methyltransferase dim-2. we identified a distinct hp1 complex, hp1, cdp-2, hda-1 and chap (hchc), and found that it is responsible for silencing ind ...201222504884
comprehensive modelling of the neurospora circadian clock and its temperature compensation.circadian clocks provide an internal measure of external time allowing organisms to anticipate and exploit predictable daily changes in the environment. rhythms driven by circadian clocks have a temperature compensated periodicity of approximately 24 hours that persists in constant conditions and can be reset by environmental time cues. computational modelling has aided our understanding of the molecular mechanisms of circadian clocks, nevertheless it remains a major challenge to integrate the l ...201222496627
organic acid excretion in penicillium ochrochloron increases with ambient ph.despite being of high biotechnological relevance, many aspects of organic acid excretion in filamentous fungi like the influence of ambient ph are still insufficiently understood. while the excretion of an individual organic acid may peak at a certain ph value, the few available studies investigating a broader range of organic acids indicate that total organic acid excretion rises with increasing external ph. we hypothesized that this phenomenon might be a general response of filamentous fungi t ...201222493592
induction of lignocellulose-degrading enzymes in neurospora crassa by cellodextrins.neurospora crassa colonizes burnt grasslands in the wild and metabolizes both cellulose and hemicellulose from plant cell walls. when switched from a favored carbon source such as sucrose to cellulose, n. crassa dramatically upregulates expression and secretion of a wide variety of genes encoding lignocellulolytic enzymes. however, the means by which n. crassa and other filamentous fungi sense the presence of cellulose in the environment remains unclear. here, we show that an n. crassa mutant ca ...201222474347
investigation of the ph-dependent electron transfer mechanism of ascomycetous class ii cellobiose dehydrogenases on electrodes.cellobiose dehydrogenase (cdh) is capable of direct electron transfer (det) on various carbon and thiol-modified gold electrodes. as a result, these systems have been utilized as biocatalyst in biosensors and biofuel cell anodes. class i cdhs, from basidiomycetous fungi, are highly specific to cellulose or lactose, and det is only observed at ph values below 5.5. to extend the applicability of cdh-based electrodes, the catalytic properties and the behavior on electrode surfaces of ascomycetous c ...201222471986
characterization of apoptosis-related oxidoreductases from neurospora crassa.the genome from neurospora crassa presented three open reading frames homologous to the genes coding for human aif and amid proteins, which are flavoproteins with oxidoreductase activities implicated in caspase-independent apoptosis. to investigate the role of these proteins, namely within the mitochondrial respiratory chain, we studied their cellular localization and characterized the respective null mutant strains. efficiency of the respiratory chain was analyzed by oxygen consumption studies ...201222470547
differential impact of nutrition on developmental and metabolic gene expression during fruiting body development in neurospora crassa.fungal fruiting body size and form are influenced by the ecology of the species, including diverse environmental stimuli. accordingly, nutritional resources available to the fungus during development can be vital to successful production of fruiting bodies. to investigate the effect of nutrition, perithecial development of neurospora crassa was induced on two different media, a chemically sparsely nutritive synthetic crossing medium (scm) and a natural carrot agar (ca). protoperithecia were coll ...201222469835
unravelling the molecular basis for light modulated cellulase gene expression - the role of photoreceptors in neurospora crassa.light represents an important environmental cue, which exerts considerable influence on the metabolism of fungi. studies with the biotechnological fungal workhorse trichoderma reesei (hypocrea jecorina) have revealed an interconnection between transcriptional regulation of cellulolytic enzymes and the light response. neurospora crassa has been used as a model organism to study light and circadian rhythm biology. we therefore investigated whether light also regulates transcriptional regulation of ...201222462823
hydrophobic motif phosphorylation coordinates activity and polar localization of the neurospora crassa nuclear dbf2-related kinase cot1.nuclear dbf2p-related (ndr) kinases and associated proteins are recognized as a conserved network that regulates eukaryotic cell polarity. ndr kinases require association with mob adaptor proteins and phosphorylation of two conserved residues in the activation segment and hydrophobic motif for activity and function. we demonstrate that the neurospora crassa ndr kinase cot1 forms inactive dimers via a conserved n-terminal extension, which is also required for the interaction of the kinase with mo ...201222451488
analysis of the role of transcription factor vad-5 in conidiation of neurospora crassa.conidiation is the major mode of reproduction in many filamentous fungi. the neurospora crassa gene vad-5, which encodes a gal4-like zn2cys6 transcription factor, was suggested to contribute to conidiation in a previous study using a knockout mutant. in this study, we confirmed the positive contribution of vad-5 to conidiation by gene complementation. to understand the role of vad-5 in conidiation, transcriptomic profiles generated by digital gene expression profiling from the vad-5 deletion mut ...201222445960
electrical phenotypes of calcium transport mutant strains of a filamentous fungus, neurospora crassa.we characterized the electrical phenotypes of mutants with mutations in genes encoding calcium transporters-a mechanosensitive channel homolog (mscs), a ca(2+)/h(+) exchange protein (cax), and ca(2+)-atpases (nca-1, nca-2, nca-3)-as well as those of double mutants (the nca-2 cax, nca-2 nca-3, and nca-3 cax mutants). the electrical characterization used dual impalements to obtain cable-corrected current-voltage measurements. only two types of mutants (the mscs mutant; the nca-2 mutant and nca-2-c ...201222408225
controlling gene expression with the q repressible binary expression system in caenorhabditis elegans.we established a transcription-based binary gene expression system in caenorhabditis elegans using the recently developed q system. this system, derived from genes in neurospora crassa, uses the transcriptional activator qf to induce the expression of target genes. activation can be efficiently suppressed by the transcriptional repressor qs, and suppression can be relieved by the nontoxic small molecule quinic acid. we used qf, qs and quinic acid to achieve temporal and spatial control of transg ...201222406855
neurospora mrc1 homologue is involved in replication stability and is required for normal cell growth and chromosome integrity in mus-9 and mus-21 mutants.stalled replication forks easily collapse and such structures can induce dna strand breaks or toxic recombination products. therefore, factors involved in stabilization of replication should be important for genome integrity. in our previous study, loss of both atm and atr homologues was shown to cause growth defects and chromosome instability in neurospora crassa. to elucidate the relationships between these defects and replication instability, we focused on one of the viable replication factor ...201222395065
sad-3, a putative helicase required for meiotic silencing by unpaired dna, interacts with other components of the silencing machinery.in neurospora crassa, genes lacking a pairing partner during meiosis are suppressed by a process known as meiotic silencing by unpaired dna (msud). to identify novel msud components, we have developed a high-throughput reverse-genetic screen for use with the n. crassa knockout library. here we describe the screening method and the characterization of a gene (sad-3) subsequently discovered. sad-3 is a putative helicase required for msud and sexual spore production. it exists in a complex with oth ...201122384347
rediscovery by whole genome sequencing: classical mutations and genome polymorphisms in neurospora crassa.classical forward genetics has been foundational to modern biology, and has been the paradigm for characterizing the role of genes in shaping phenotypes for decades. in recent years, reverse genetics has been used to identify the functions of genes, via the intentional introduction of variation and subsequent evaluation in physiological, molecular, and even population contexts. these approaches are complementary and whole genome analysis serves as a bridge between the two. we report in this arti ...201122384341
a fluorescence-based supramolecular tandem assay for monitoring lysine methyltransferase activity in homogeneous solution.the demand for practical and convenient enzyme assays for histone lysine methyltransferases (hkmts) emerges along with the rapid development of this young class of enzymes. a supramolecular reporter pair composed of p-sulfonatocalix[4]arene (cx4) and the fluorescent dye lucigenin (lcg) has been used to monitor enzymatic trimethylation of lysine residues in peptide substrates. the assay affords a switch-on fluorescence response and operates in a continuous, real-time, and label-free fashion. the ...201222367964
temperature-sensitive and circadian oscillators of neurospora crassa share components.in neurospora crassa, the interactions between products of the frequency (frq), frequency-interacting rna helicase (frh), white collar-1 (wc-1), and white collar-2 (wc-2) genes establish a molecular circadian clockwork, called the frq-wc-oscillator (fwo), which is required for the generation of molecular and overt circadian rhythmicity. in strains carrying nonfunctional frq alleles, circadian rhythms in asexual spore development (conidiation) are abolished in constant conditions, yet conidiation ...201222367035
deciphering transcriptional regulatory mechanisms associated with hemicellulose degradation in neurospora crassa.hemicellulose, the second most abundant plant biomass fraction after cellulose, is widely viewed as a potential substrate for the production of liquid fuels and other value-added materials. degradation of hemicellulose by filamentous fungi requires production of many different enzymes, which are induced by biopolymers or its derivatives and regulated mainly at the transcriptional level through transcription factors (tfs). neurospora crassa, a model filamentous fungus, expresses and secretes enzy ...201222345350
genome-wide selection on codon usage at the population level in the fungal model organism neurospora crassa.many organisms exhibit biased codon usage in their genome, including the fungal model organism neurospora crassa. the preferential use of subset of synonymous codons (optimal codons) at the macroevolutionary level is believed to result from a history of selection to promote translational efficiency. at present, few data are available about selection on optimal codons at the microevolutionary scale, that is, at the population level. herein, we conducted a large-scale assessment of codon mutations ...201222334579
selection of a dna barcode for nectriaceae from fungal whole-genomes.a dna barcode is a short segment of sequence that is able to distinguish species. a barcode must ideally contain enough variation to distinguish every individual species and be easily obtained. fungi of nectriaceae are economically important and show high species diversity. to establish a standard dna barcode for this group of fungi, the genomes of neurospora crassa and 30 other filamentous fungi were compared. the expect value was treated as a criterion to recognize homologous sequences. four c ...201222314494
self-assembly of functional, amphipathic amyloid monolayers by the fungal hydrophobin eas.the hydrophobin eas from the fungus neurospora crassa forms functional amyloid fibrils called rodlets that facilitate spore formation and dispersal. self-assembly of eas into fibrillar rodlets occurs spontaneously at hydrophobic:hydrophilic interfaces and the rodlets further associate laterally to form amphipathic monolayers. we have used site-directed mutagenesis and peptide experiments to identify the region of eas that drives intermolecular association and formation of the cross-β rodlet stru ...201222308366
[analysis of several key problems about the two linkage gene mapping of neurospora crassa in genetics teaching].ordered tetrad analysis is important content in the genetics teaching. in particular, the two linkage gene mapping is not only a key point, but also a difficult one. how to explain the content better is a hard nut for the many genetics teachers or editors of the teaching material to crack. here, based on teaching practice of many years we summarized several key problems, which are difficult to understand by students and frequently neglected by the teachers and the editors of genetics. furthermor ...201222306881
roles for receptors, pheromones, g proteins, and mating type genes during sexual reproduction in neurospora crassa.here we characterize the relationship between the pre-2 pheromone receptor and its ligand, ccg-4, and the general requirements for receptors, pheromones, g proteins, and mating type genes during fusion of opposite mating-type cells and sexual sporulation in the multicellular fungus neurospora crassa. pre-2 is highly expressed in mat a cells and is localized in male and female reproductive structures. δpre-2 mat a females do not respond chemotropically to mat a males (conidia) or form mature frui ...201222298702
comparative live-cell imaging analyses of spa-2, bud-6 and bni-1 in neurospora crassa reveal novel features of the filamentous fungal polarisome.a key multiprotein complex involved in regulating the actin cytoskeleton and secretory machinery required for polarized growth in fungi, is the polarisome. recognized core constituents in budding yeast are the proteins spa2, pea2, aip3/bud6, and the key effector bni1. multicellular fungi display a more complex polarized morphogenesis than yeasts, suggesting that the filamentous fungal polarisome might fulfill additional functions. in this study, we compared the subcellular organization and dynam ...201222291944
excitable behavior can explain the "ping-pong" mode of communication between cells using the same chemoattractant.here we elucidate a paradox: how a single chemoattractant-receptor system in two individuals is used for communication despite the seeming inevitability of self-excitation. in the filamentous fungus neurospora crassa, genetically identical cells that produce the same chemoattractant fuse via the homing of individual cell protrusions toward each other. this is achieved via a recently described "ping-pong" pulsatile communication. using a generic activator-inhibitor model of excitable behavior, we ...201222271443
cooperation among germinating spores facilitates the growth of the fungus, neurospora crassa.fusions between individuals are a common feature of organisms with modular, indeterminate life forms, including plants, marine invertebrates and fungi. the consequences of fusion for an individual fungus are poorly understood. we used wild-type and fusion mutant strains of the genetic model neurospora crassa to chronicle the fitness in two different laboratory habitats, and in each experiment started colonies from multiple different densities of asexual spores. on round petri dishes, fusion enab ...201222258449
substitutions in the amino-terminal tail of neurospora histone h3 have varied effects on dna methylation.eukaryotic genomes are partitioned into active and inactive domains called euchromatin and heterochromatin, respectively. in neurospora crassa, heterochromatin formation requires methylation of histone h3 at lysine 9 (h3k9) by the set domain protein dim-5. heterochromatin protein 1 (hp1) reads this mark and directly recruits the dna methyltransferase, dim-2. an ectopic h3 gene carrying a substitution at k9 (hh3(k9l) or hh3(k9r)) causes global loss of dna methylation in the presence of wild-type ...201122242002
a crossover hotspot near his-3 in neurospora crassa is a preferential recombination termination site.during analysis of 148 unselected neurospora crassa octads, an above average rate of crossing over was detected within a 360-base region near the 3' end of his-3, suggesting a hotspot for crossing over about 1.8 kb away from the recombination initiation site within cog. homozygous deletion of the 360-base region increases exchanges in his-3 and on the far side of his-3 from cog, with the heterozygote showing an intermediate increase. we conclude that recombination events initiated at cog termina ...201222203161
oxidative cleavage of cellulose by fungal copper-dependent polysaccharide monooxygenases.fungal-derived, copper-dependent polysaccharide monooxygenases (pmos), formerly known as gh61 proteins, have recently been shown to catalyze the o(2)-dependent oxidative cleavage of recalcitrant polysaccharides. different pmos isolated from neurospora crassa were found to generate oxidized cellodextrins modified at the reducing or nonreducing ends upon incubation with cellulose and cellobiose dehydrogenase. here we show that the nonreducing end product formed by an n. crassa pmo is a 4-ketoaldos ...201222188218
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