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post-translational modifications in circadian rhythms.the pace has quickened in circadian biology research. in particular, an abundance of results focused on post-translational modifications (ptms) is sharpening our view of circadian molecular clockworks. ptms affect nearly all aspects of clock biology; in some cases they are essential for clock function and in others, they provide layers of regulatory fine-tuning. our goal is to review recent advances in clock ptms, help make sense of emerging themes, and spotlight intriguing (and perhaps controve ...200919740663
optimization of coprogen production in neurospora crassa.coprogen production of neurospora crassa was dependent on glucose, aspartate and iron contents as well as on initial ph of the culture media. surplus iron and acidic ph hindered the production of coprogen as well as the transcription of the sid1 gene (ncu07117) encoding putative l-ornithine-n5-monooxygenase, the first enzyme in the coprogen biosynthetic pathway. high glucose (40 g/l) and aspartate (21 g/l) concentrations were beneficial for coprogen synthesis, but neither glucose nor aspartate a ...200919700391
transcriptional profiling and functional analysis of heterokaryon incompatibility in neurospora crassa reveals that reactive oxygen species, but not metacaspases, are associated with programmed cell death.heterokaryon incompatibility (hi) is a nonself recognition phenomenon occurring in filamentous fungi that is important for limiting resource plundering and restricting viral transfer between strains. nonself recognition and hi occurs during hyphal fusion between strains that differ at het loci. if two strains undergo hyphal fusion, but differ in allelic specificity at a het locus, the fusion cell is compartmentalized and undergoes a rapid programmed cell death (pcd). incompatible heterokaryons s ...200919696111
cytoplasmic bulk flow propels nuclei in mature hyphae of neurospora crassa.we used confocal microscopy to evaluate nuclear dynamics in mature, growing hyphae of neurospora crassa whose nuclei expressed histone h1-tagged green fluorescent protein (gfp). in addition to the h1-gfp wild-type (wt) strain, we examined nuclear displacement (passive transport) in four mutants deficient in microtubule-related motor proteins (ro-1, ro-3, kin-1, and a ro-1 kin-1 double mutant). we also treated the wt strain with benomyl and cytochalasin a to disrupt microtubules and actin microfi ...200919684281
kinetic evidence for an anion binding pocket in the active site of nitronate monooxygenase.a series of monovalent, inorganic anions and aliphatic aldehydes were tested as inhibitors for hansenula mrakii and neurospora crassa nitronate monooxygenase, formerly known as 2-nitropropane dioxygenase, to investigate the structural features that contribute to the binding of the anionic nitronate substrates to the enzymes. a linear correlation between the volumes of the inorganic anions and their effectiveness as competitive inhibitors of the enzymes was observed in a plot of pk(is)versus the ...200919683782
rhythmic conidiation in constant light in vivid mutants of neurospora crassa.in neurospora crassa, a circadian rhythm of conidiation (asexual spore formation) can be seen on the surface of agar media. this rhythm has a period of 22 hr in constant darkness (d/d). under constant illumination (l/l), no rhythm is visible and cultures show constant conidiation. however, here we report that strains with a mutation in the vivid (vvd) gene, previously shown to code for the photoreceptor involved in photo-adaptation, exhibit conidiation rhythms in l/l as well as in d/d. the perio ...200919139144
characterization and mrna expression profile of the tbnre1 gene of the ectomycorrhizal fungus tuber borchii.this study focuses on the cloning and characterization of the major nitrogen regulator element from the ectomycorrhizal fungus tuber borchii, tbnre1. sequence analysis of the predicted protein and complementation experiments in neurospora crassa demonstrated that the cloned gene is orthologous to area/nit-2 gene. transcriptional expression investigations by real-time rt-pcr showed tbnre1 up-regulation in the presence of nitrate or in the absence of nitrogen during free-living mycelium growth. on ...200919115055
characterization of chromosome ends in the filamentous fungus neurospora crassa.telomeres and subtelomere regions have vital roles in cellular homeostasis and can facilitate niche adaptation. however, information on telomere/subtelomere structure is still limited to a small number of organisms. prior to initiation of this project, the neurospora crassa genome assembly contained only 3 of the 14 telomeres. the missing telomeres were identified through bioinformatic mining of raw sequence data from the genome project and from clones in new cosmid and plasmid libraries. their ...200919104079
relics of repeat-induced point mutation direct heterochromatin formation in neurospora crassa.both rnai-dependent and -independent mechanisms have been implicated in the establishment of heterochromatin domains, which may be stabilized by feedback loops involving chromatin proteins and modifications of histones and dna. neurospora crassa sports features of heterochromatin found in higher eukaryotes, namely cytosine methylation (5mc), methylation of histone h3 lysine 9 (h3k9me), and heterochromatin protein 1 (hp1), and is a model to investigate heterochromatin establishment and maintenanc ...200919092133
transcriptional analysis of the response of neurospora crassa to phytosphingosine reveals links to mitochondrial function.treatment of neurospora crassa cells with phytosphingosine (phs) induces programmed cell death (pcd) by an unknown mechanism. to determine the relationship between phs treatment and pcd, we determined changes in global gene expression levels in n. crassa during a time-course of phs treatment. most genes having differential expression levels compared to untreated samples showed an increase in relative expression level upon phs exposure. however, genes encoding mitochondrial proteins were highly e ...200919520721
relationship between phylogenetic distribution and genomic features in neurospora crassa.in the post-genome era, insufficient functional annotation of predicted genes greatly restricts the potential of mining genome data. we demonstrate that an evolutionary approach, which is independent of functional annotation, has great potential as a tool for genome analysis. we chose the genome of a model filamentous fungus neurospora crassa as an example. phylogenetic distribution of each predicted protein coding gene (pcg) in the n. crassa genome was used to classify genes into six mutually e ...200919461939
qirna is a new type of small interfering rna induced by dna damage.rna interference pathways use small rnas to mediate gene silencing in eukaryotes. in addition to small interfering rnas (sirnas) and micrornas, several types of endogenously produced small rnas have important roles in gene regulation, germ cell maintenance and transposon silencing. the production of some of these rnas requires the synthesis of aberrant rnas (arnas) or pre-sirnas, which are specifically recognized by rna-dependent rna polymerases to make double-stranded rna. the mechanism for arn ...200919444217
activity of the circadian transcription factor white collar complex is modulated by phosphorylation of sp-motifs.posttranslational modifications, particularly phosphorylation, regulate activity, stability and localization of proteins in circadian clocks, thereby contributing to a stable oscillation with a period of approximately 24h. the white collar complex (wcc) is the central transcription factor of the circadian clock of neurospora crassa. its activity is regulated in a circadian manner by rhythmic phosphorylation, mediated by the clock protein frequency (frq). here we present purification of tap-tagge ...200919427309
minimum criteria for dna damage-induced phase advances in circadian rhythms.robust oscillatory behaviors are common features of circadian and cell cycle rhythms. these cyclic processes, however, behave distinctively in terms of their periods and phases in response to external influences such as light, temperature, nutrients, etc. nevertheless, several links have been found between these two oscillators. cell division cycles gated by the circadian clock have been observed since the late 1950s. on the other hand, ionizing radiation (ir) treatments cause cells to undergo a ...200919424508
neurospora crassa fmf-1 encodes the homologue of the schizosaccharomyces pombe ste11p regulator of sexual development.the neurospora crassa fmf-1 mutation exerts an unusual 'perithecium-dominant' developmental arrest; fmf-1 x fmf-1+ cross becomes arrested in perithecial development regardless of whether the mutant participates in the cross as the male or female parent. we localized fmf-1 to the lg il genome segment between the centromere-proximal breakpoint of the chromosome segment duplication dp(il)39311 and the centromere. by mapping crossovers with respect to rflp markers in this region we further localized ...200919417542
molecular view of the interaction between iota-carrageenan and a phospholipid film and its role in enzyme immobilization.proteins incorporated into phospholipid langmuir-blodgett (lb) films are a good model system for biomembranes and enzyme immobilization studies. the specific fluidity of biomembranes, an important requisite for enzymatic activity, is naturally controlled by varying phospholipid compositions. in a model system, instead, lb film fluidity may be varied by covering the top layer with different substances able to interact simultaneously with the phospholipid and the protein to be immobilized. in this ...200919415915
chitosan permeabilizes the plasma membrane and kills cells of neurospora crassa in an energy dependent manner.chitosan has been reported to inhibit spore germination and mycelial growth in plant pathogens, but its mode of antifungal action is poorly understood. following chitosan treatment, we characterized plasma membrane permeabilization, and cell death and lysis in the experimental model, neurospora crassa. rhodamine-labeled chitosan was used to show that chitosan is internalized by fungal cells. cell viability stains and the calcium reporter, aequorin, were used to monitor plasma membrane permeabili ...200919389478
a calcium binding protein from cell wall of neurospora crassa.isolated cell wall preparations of n. crassa bind significant levels of ca, mg and other divalent cations. enzymatic treatment of the cell wall with beta-(1,3)-glucanase, but not with chitinase, resulted in solubilization of only the calcium-binding protein fraction. a calcium-binding protein (cabp) was purified by metal-chelate affinity chromatography and reversed phase hplc. cabp has an mr of around 6 kda on sds-page and mass spectrometry showed that it has a molecular mass of 5744 da. one mol ...200919322841
vaccine production in neurospora crassa.we have chosen to use the filamentous fungus neurospora crassa to produce subunit vaccines. here we describe the production and purification of influenza hemagglutinin and neuraminidase antigens in n. crassa. the n. crassa system used by neugenesis offers many advantages over other systems for production of recombinant protein. in contrast to mammalian cell culture, n. crassa can be grown in a rapid and economic manner, generating large amounts of recombinant protein in simple, defined medium. v ...200919285427
a novel polyketide biosynthesis gene cluster is involved in fruiting body morphogenesis in the filamentous fungi sordaria macrospora and neurospora crassa.during fungal fruiting body development, hyphae aggregate to form multicellular structures that protect and disperse the sexual spores. analysis of microarray data revealed a gene cluster strongly upregulated during fruiting body development in the ascomycete sordaria macrospora. real time pcr analysis showed that the genes from the orthologous cluster in neurospora crassa are also upregulated during development. the cluster encodes putative polyketide biosynthesis enzymes, including a reducing ...200919277664
genome-wide analysis of light-inducible responses reveals hierarchical light signalling in neurospora.white collar-1 (wc-1) and white collar-2 (wc-2) are essential for light-mediated responses in neurospora crassa, but the molecular mechanisms underlying gene induction and the roles of other real and putative photoreceptors remain poorly characterized. unsupervised hierarchical clustering of genome-wide microarrays reveals 5.6% of detectable transcripts, including several novel mediators, that are either early or late light responsive. evidence is shown for photoreception in the absence of the d ...200919262566
quelling targets the rdna locus and functions in rdna copy number control.rna silencing occurs in a broad range of organisms. although its ancestral function is probably related to the genome defense mechanism against repetitive selfish elements, it has been found that rna silencing regulates different cellular processes such as gene expression and chromosomal segregation. in neurospora crassa, a rna silencing mechanism, called quelling, acts to repress the expression of transgenes and transposons, but until now no other cellular functions have been shown to be regula ...200919243581
effects of mitochondrial complex iii disruption in the respiratory chain of neurospora crassa.in mitochondria from most organisms, including neurospora crassa, dimeric complex iii was found associated with complex i. additional association of complex iv with this core structure leads to the formation of a respirasome. it was recently described for bacteria and mammals that complex iii is needed for the assembly/stability of complex i. to elucidate the role of complex iii in the organization of the respiratory chain of n. crassa, we analysed strains devoid of either the rieske iron-sulphu ...200919239619
the bem46-like protein appears to be essential for hyphal development upon ascospore germination in neurospora crassa and is targeted to the endoplasmic reticulum.the bud emergence (bem)46 proteins are evolutionarily conserved members of the alpha/beta-hydrolase super family, but their exact role remains unknown. to better understand the cellular role of bem46 and its homologs, we used the model organism neurospora crassa in conjunction with bem46 rnai, over-expression vectors, and repeat induced point mutation analyzes. we clearly demonstrated that bem46 is required for cell type-specific hyphal growth, which indicates a role for bem46 in maintaining pol ...200919238386
catabolite repression directly affects transcription of the qa-y gene of neurospora crassa.the quinic acid (qa) gene cluster of neurospora crassa is subject to two levels of gene control: a primary system which responds to the presence of quinic acid via the qa-1s repressor protein, and a secondary system which represses transcription of qa genes in the presence of a preferred carbon source, e.g. glucose. rna blot analysis revealed that transcription of the qa-y gene, which encodes a quinic acid permease, was strongly repressed in the presence of glucose even in the absence of the qa- ...200919236936
transcriptional changes in the nuc-2a mutant strain of neurospora crassa cultivated under conditions of phosphate shortage.the molecular mechanism that controls the response to phosphate shortage in neurospora crassa involves four regulatory genes -nuc-2, preg, pgov, and nuc-1. phosphate shortage is sensed by the nuc-2 gene, the product of which inhibits the functioning of the preg-pgov complex. this allows the translocation of the transcriptional factor nuc-1 into the nucleus, which activates the transcription of phosphate-repressible phosphatases. the nuc-2a mutant strain of n. crassa carries a loss-of-function mu ...200919230635
chromosome segment duplications in neurospora crassa: barren crosses beget fertile science.studies on neurospora chromosome segment duplications (dps) performed since the publication of perkins's comprehensive review in 1997 form the focus of this article. we present a brief summary of perkins's seminal work on chromosome rearrangements, specifically, the identification of insertional and quasiterminal translocations that can segregate dp progeny when crossed with normal sequence strains (i.e., t x n). we describe the genome defense process called meiotic silencing by unpaired dna tha ...200919204993
a sorghum 85kda heat stress-modulated protein shows calmodulin-binding properties and cross-reactivity to anti-neurospora crassa hsp 80 antibodies.the present study, carried out to identify stress-modulated calmodulin (cam)-binding proteins in sorghum, resulted in the isolation of several proteins, which showed binding to cam-sepharose matrix. calmodulin gel overlay assay and maldi-tof ms analysis revealed that an 85kda protein (hsp85), which interacted with calmodulin, cross-reacted with anti-n. crassa hsp80 antibodies. since these antibodies bind to plant hsp90, sorghum hsp85 is likely to be a member of the hsp90 family. this study provi ...200919174162
systems analysis of plant cell wall degradation by the model filamentous fungus neurospora crassa.the filamentous fungus neurospora crassa is a model laboratory organism, but in nature is commonly found growing on dead plant material, particularly grasses. using functional genomics resources available for n. crassa, which include a near-full genome deletion strain set and whole genome microarrays, we undertook a system-wide analysis of plant cell wall and cellulose degradation. we identified approximately 770 genes that showed expression differences when n. crassa was cultured on ground misc ...200920018766
neurospora sees the light: light signaling components in a model system.light is a key environmental signal for most life on earth. over 5% of neurospora crassa genes are expressed in response to light stimulation in a temporally regulated cascade that includes several transcription factors. fungal genomes, including neurospora's, may encode several different proteins capable of binding chromophores with the ability to harvest light energy as well as proteins that can interact with primary photoreceptors or further propogate the light signal. the best understood pho ...200919907715
snornas from the filamentous fungus neurospora crassa: structural, functional and evolutionary insights.snornas represent an excellent model for studying the structural and functional evolution of small non-coding rnas involved in the post-transcriptional modification machinery for rrnas and snrnas in eukaryotic cells. identification of snornas from neurospora crassa, an important model organism playing key roles in the development of modern genetics, biochemistry and molecular biology will provide insights into the evolution of snorna genes in the fungus kingdom.200919895704
oscillatory recruitment of signaling proteins to cell tips promotes coordinated behavior during cell fusion.cell-cell communication is essential for coordinating physiological responses in multicellular organisms and is required for various developmental processes, including cell migration, differentiation, and fusion. to facilitate communication, functional differences are usually required between interacting cells, which can be established either genetically or developmentally. however, genetically identical cells in the same developmental state are also capable of communicating, but must avoid self ...200919884508
functional stratification of the spitzenkörper of neurospora crassa.gs-1 (ncu04189) is a protein required for the synthesis of beta-1,3-glucan in neurospora crassa. as chitin, beta-1,3-glucan is a morphogenetically relevant component of the fungal cell wall. previously, we showed that chitin synthases are delivered to the growing hyphal tip of n. crassa by secretory microvesicles that follow an unconventional route and accumulate in the core of the spitzenkörper (spk). tagged with the green fluorescent protein (gfp), gs-1 accumulated in the hyphal apex forming a ...200919843220
cell elongation and branching are regulated by differential phosphorylation states of the nuclear dbf2-related kinase cot1 in neurospora crassa.dysfunction of the neurospora crassa nuclear dbf2-related kinase cot1 leads to cessation of tip extension and massive induction of new sites of growth. to determine the role phosphorylation plays in cot1 function, we mutated cot1 residues corresponding to positions of highly conserved nuclear dbf2-related phosphorylation sites. analyses of the point-mutation cot-1 strains (mimicking non- and constitutively phosphorylated states) indicate the involvement of cot1 phosphorylation in the regulation ...200919818014
genome-wide high throughput analysis of dna methylation in eukaryotes.cytosine methylation is the quintessential epigenetic mark. two well-established methods, bisulfite sequencing and methyl-dna immunoprecipitation (medip) lend themselves to the genome-wide analysis of dna methylation by high throughput sequencing. here we provide an overview and brief review of these methods. we summarize our experience with medip followed by high throughput illumina/solexa sequencing, exemplified by the analysis of the methylated fraction of the neurospora crassa genome ("methy ...200918950712
plasticity of a key metabolic pathway in fungi.beta oxidation is the principal metabolic pathway for fatty acid degradation. the pathway is virtually universally present throughout eukaryotes yet displays different forms in enzyme architecture, substrate specificity, and subcellular location. in this review, we examine beta oxidation across the fungal kingdom by conducting a large-scale in silico screen and localization prediction for all relevant enzymes in >50 species. the survey reveals that fungi exhibit an astounding diversity of beta o ...200918795352
a mutation within the catalytic domain of cot1 kinase confers changes in the presence of two cot1 isoforms and in ser/thr protein kinase and phosphatase activities in neurospora crassa.neurospora crassa grows by forming spreading colonies. cot-1 belongs to a class of n. crassa colonial temperature-sensitive (cot) mutants and encodes a ser/thr protein kinase. we have mapped the cot-1 mutation to a single base change resulting in a his to arg substitution at amino acid 351, which resides within the catalytic domain. antibodies raised against cot1 detected and immunoprecipitated a predominant 73-kda polypeptide in n. crassa extracts, whose abundance was constant under all growth ...200910441452
role of cytochrome c heme lyase in the import of cytochrome c into mitochondria.the import of cytochrome c into neurospora crassa mitochondria was examined at distinct stages in vitro. the precursor protein, apocytochrome c, binds to mitochondria with high affinity and specificity but is not transported completely across the outer membrane in the absence of conversion to holocytochrome c. the bound apocytochrome c is accessible to externally added proteases but at the same time penetrates far enough through the outer membrane to interact with cytochrome c heme lyase. format ...20092848813
negative roles of a novel nitrogen metabolite repression-related gene, tar1, in laccase production and nitrate utilization by the basidiomycete cryptococcus neoformans.the multicopper oxidase laccase is widespread in fungi and has great industrial importance. one puzzle regarding laccase production in the basidiomycetous yeast cryptococcus neoformans is that it is inhibited by high temperature (e.g., 37 degrees c). in this paper, we report identification of a nitrogen metabolite repression-related gene, tar1, which is responsible for laccase repression. disruption of tar1 results in a significant increase in the level of lac1 mrna at 37 degrees c. the putative ...200919734333
nucleoside diphosphate kinase-1 regulates hyphal development via the transcriptional regulation of catalase in neurospora crassa.a nucleoside diphosphate kinase-1-disrupted (ndk-1(rip-1)) mutant was observed to be defective in aerial hyphal and conidial development. in this study, two types of hyphae, fine and thick, were observed in wild-type (wt) strains. however, only fine-type hyphae were observed in the ndk-1(rip-1) mutants. the ndk-1(rip-1) mutants were stimulated by oxidative stress and constitutively expressed an antioxidant enzyme catalase (cat)-3. furthermore the ndk-1(rip-1) mutants could form thick hyphae by o ...200919765588
distinct changes in soybean xylem sap proteome in response to pathogenic and symbiotic microbe interactions.plant systemic signaling characterized by the long distance transport of molecules across plant organs involves the xylem and phloem conduits. root-microbe interactions generate systemic signals that are transported to aerial organs via the xylem sap. we analyzed the xylem sap proteome of soybean seedlings in response to pathogenic and symbiotic interactions to identify systemic signaling proteins and other differentially expressed proteins.200919772575
ptk2 contributes to osmoadaptation in the filamentous fungus neurospora crassa.hyphal tip-growing organisms often rely upon an internal hydrostatic pressure (turgor) to drive localized expansion of the cell. regulation of the turgor in response to osmotic shock is mediated primarily by an osmotic map kinase cascade which activates osmolyte synthesis and ion uptake to effect turgor recovery. we characterized a neurospora crassa homolog (ptk2) of ser/thr kinase regulators of ion transport in yeast to determine its role in turgor regulation in a filamentous fungi. the ptk2 mu ...200919772928
two ndr kinase-mob complexes function as distinct modules during septum formation and tip extension in neurospora crassa.ndr kinases are important for growth and differentiation and require interaction with mob proteins for activity and function. we characterized the ndr kinases and mob activators in neurospora crassa and identified two ndr kinases (cot1 and dbf2) and four mob proteins (mob1, mob2a, mob2b and mob3/phocein) that form two functional ndr-mob protein complexes. the mob1-dbf2 complex is not only essential for septum formation in vegetative cells and during conidiation, but also functions during sexual ...200919788544
structure and distribution of organelles and cellular location of calcium transporters in neurospora crassa.we wanted to examine the cellular locations of four neurospora crassa proteins that transport calcium. however, the structure and distribution of organelles in live hyphae of n. crassa have not been comprehensively described. therefore, we made recombinant genes that generate translational fusions of putative organellar marker proteins with green or red fluorescent protein. we observed putative endoplasmic reticulum proteins, encoded by grp-78 and dpm, in the nuclear envelope and associated memb ...200919801418
self-signalling and self-fusion in filamentous fungi.the formation of interconnected hyphal networks is central to the organisation and functioning of the filamentous fungal colony. it is brought about by the fusion of specialised hyphae during colony initiation and mature colony development. these hyphae are normally genetically identical, and hence this process is termed hyphal self-fusion. the conidial anastomosis tube (cat) functions in forming networks of conidial germlings during colony initiation. this hyphal type in neurospora crassa is be ...200919864177
observing meiosis in filamentous fungi: sordaria and neurospora.the filamentous fungi neurospora crassa and sordaria macrospora are materials of choice for recombination studies because each of the dna strands involved in meiosis can be visually analyzed using spore-color mutants. well-advanced molecular genetic methodologies have been developed for each of these fungi, and several mutants defective in recombination and/or pairing are available. moreover, the complete genome sequence of n. crassa has made it possible to clone virtually any gene involved in t ...200919685321
hydrothermal processing and enzymatic hydrolysis of sorghum bagasse for fermentable carbohydrates production.untreated and hydrothermally treated sorghum bagasse (sb) was hydrolyzed to simple sugars by the synergistic action of cellulases and hemicellulases produced by the fungi fusarium oxysporum and neurospora crassa. synergism between the two lignocellulolytic systems was maximized with the application of higher fraction of n. crassa enzymes. hydrothermolysis of sb was studied at a wide range of treatment times and temperatures. at intense pretreatment conditions (210 degrees c for 20 min; logr(0)=4 ...200919692234
synthesis and antifungal activity of beta-trifluoroalkyl aminovinyl ketone derivatives.ten beta-trifluoroalkyl aminovinyl ketone derivatives were synthesized, and their inhibitory effects on several phytopathogenic fungi, an oomycete and plants were assessed. the various compounds were fungitoxic at the 10-100 microm range, with (z)-3-amino-4,4,4-trifluoro-1-(4-chlorophenyl)but-2-en-1-one exhibiting the highest inhibitory effect on most of the test pathogens. alternaria alternata and neurospora crassa were the most tolerant and sensitive fungi to the compounds, respectively. we pr ...200919711891
[application of bioinformatics analysis of signal peptide in the identification of neurospora crassa phya gene].to identify the function of the gene encoding neurospora crassa eaa33149.1 protein which has 46.85% similarity with aspergillus niger pha gene.200919726332
evolutionary roles of upstream open reading frames in mediating gene regulation in fungi.upstream open reading frames (uorfs) are frequently present in the 5'-leader regions of fungal mrnas. they can affect translation by controlling the ability of ribosomes that scan from the mrna 5' end to reach the downstream genic reading frame. the translation of uorfs can also affect mrna stability. for several genes, including saccharomyces cerevisiae gcn4, s. cerevisiae cpa1, and neurospora crassa arg-2, regulation by uorfs controls expression in response to specific physiological signals. t ...200919514854
trifluoromethanesulfonic acid-based proteomic analysis of cell wall and secreted proteins of the ascomycetous fungi neurospora crassa and candida albicans.cell wall proteins from purified candida albicans and neurospora crassa cell walls were released using trifluoromethanesulfonic acid (tfms) which cleaves the cell wall glucan/chitin matrix and deglycosylates the proteins. the cell wall proteins were then characterized by sds-page and identified by proteomic analysis. the analyses for c. albicans identified 15 cell wall proteins and six secreted proteins. for n. crassa, the analyses identified 26 cell wall proteins and nine secreted proteins. mos ...200919555771
enzymatic synthesis of bis-5-alkylresorcinols by resorcinol-producing type iii polyketide synthases.no enzyme systems responsible for the biosynthesis of structurally and biosynthetically intriguing bis-5-alkylresorcinols produced by plants have been identified. herein, we show that bacterial, fungal and plant alkylresorcinol-producing type iii polyketide synthases (pkss), such as arsb in the gram-negative bacterium azotobacter vinelandii, oras in the fungus neurospora crassa and aras2 in the rice plant oryza sativa, can synthesize bis-5-alkylresorcinol from alkanedioic acid n-acetylcysteamine ...200919557027
cell-free protein synthesis energized by slowly-metabolized maltodextrin.cell-free protein synthesis (cfps) is a rapid and high throughput technology for obtaining proteins from their genes. the primary energy source atp is regenerated from the secondary energy source through substrate phosphorylation in cfps.200919558718
deletion and overexpression of the aspergillus nidulans gata factor areb reveals unexpected pleiotropy.the aspergillus nidulans transcription factor area is a key regulator of nitrogen metabolic gene expression. area contains a c-terminal gata zinc finger dna-binding domain and activates expression of genes necessary for nitrogen acquisition. previous studies identified areb as a potential negative regulator of nitrogen catabolism showing similarity with penicillium chrysogenum nreb and neurospora crassa asd4. the areb gene encodes multiple products containing an n-terminal gata zinc finger and a ...200919628561
the het-c heterokaryon incompatibility gene in aspergillus niger.heterokaryon incompatibility among aspergillus niger strains is a widespread phenomenon that is observed as the inability to form stable heterokaryons. the genetic basis of heterokaryon incompatibility reactions is well established in some sexual filamentous fungi but largely unknown in presumed asexual species, such as a. niger. to test whether the genes that determine heterokaryon incompatibility in neurospora crassa, such as het-c, vib-1 and pin-c, have a similar function in a. niger, we perf ...200919015029
the saccharomyces cerevisiae prm1 homolog in neurospora crassa is involved in vegetative and sexual cell fusion events but also has postfertilization functions.cell-cell fusion is essential for a variety of developmental steps in many eukaryotic organisms, during both fertilization and vegetative cell growth. although the molecular mechanisms associated with intracellular membrane fusion are well characterized, the molecular mechanisms of plasma membrane merger between cells are poorly understood. in the filamentous fungus neurospora crassa, cell fusion events occur during both vegetative and sexual stages of its life cycle, thus making it an attractiv ...200919064710
heterologous expression in escherichia coli of neurospora crassa neutral trehalase as an active enzyme.neutral trehalase from neurospora crassa was expressed in escherichia coli as a polypeptide of approximately 84 kda in agreement with the theoretical size calculated from the corresponding cdna. the recombinant neutral trehalase, purified by affinity chromatography exhibited a specific activity of 80-150 mu/mg protein. optima of ph and temperature were 7.0 and 30 degrees c, respectively. the enzyme was absolutely specific for trehalose, and was quite sensitive to incubation at 40 degrees c. the ...200919073263
structure-function relationships in fungal large-subunit catalases.neurospora crassa has two large-subunit catalases, cat-1 and cat-3. cat-1 is associated with non-growing cells and accumulates particularly in asexual spores; cat-3 is associated with growing cells and is induced under different stress conditions. it is our interest to elucidate the structure-function relationships in large-subunit catalases. here we have determined the cat-3 crystal structure and compared it with the previously determined cat-1 structure. similar to cat-1, cat-3 hydrogen peroxi ...200919109972
telomere organization in the ligninolytic basidiomycete pleurotus ostreatus.telomeres are structural and functional chromosome regions that are essential for the cell cycle to proceed normally. they are, however, difficult to map genetically and to identify in genome-wide sequence programs because of their structure and repetitive nature. we studied the telomeric and subtelomeric organization in the basidiomycete pleurotus ostreatus using a combination of molecular and bioinformatics tools that permitted us to determine 19 out of the 22 telomeres expected in this fungus ...200919114509
dimerization of the yeast eukaryotic translation initiation factor 5a requires hypusine and is rna dependent.post-translational modification of the highly conserved k51 residue of the saccharomyces cerevisiae eukaryotic translation initiation factor 5a (eif5a) to form hypusine, is essential for its many functions including the binding of specific mrnas. we characterized hypusinated yeast eif5a by size-exclusion chromatography and native page, showing that the protein exists as a homodimer. a k51r mutant, which was not functional in vivo eluted as a monomer and inhibition of hypusination abolished dimer ...200919120453
understanding the plasticity of the alpha/beta hydrolase fold: lid swapping on the candida antarctica lipase b results in chimeras with interesting biocatalytic properties.the best of both worlds. long molecular dynamics (md) simulations of candida antarctica lipase b (calb) confirmed the function of helix alpha5 as a lid structure. replacement of the helix with corresponding lid regions from calb homologues from neurospora crassa and gibberella zeae resulted in new calb chimeras with novel biocatalytic properties. the figure shows a snapshot from the md simulation. the candida antarctica lipase b (calb) has found very extensive use in biocatalysis reactions. long ...200919156649
tools for fungal proteomics: multifunctional neurospora vectors for gene replacement, protein expression and protein purification.the completion of genome-sequencing projects for a number of fungi set the stage for detailed investigations of proteins. we report the generation of versatile expression vectors for detection and isolation of proteins and protein complexes in the filamentous fungus neurospora crassa. the vectors, which can be adapted for other fungi, contain c- or n-terminal flag, ha, myc, gfp, or hat-flag epitope tags with a flexible poly-glycine linker and include sequences for targeting to the his-3 locus in ...200919171944
inflated kinetic isotope effects in the branched mechanism of neurospora crassa 2-nitropropane dioxygenase.catalytic turnover of neurospora crassa 2-nitropropane dioxygenase with nitroethane as substrate occurs through both nonoxidative and oxidative pathways. the ph dependence of the kinetic isotope effects with [1,1-(2)h(2)]nitroethane as substrate was measured in the current study by monitoring the formation of the nitronate product in the nonoxidative pathway. the kinetic isotope effect on the second-order rate constant for nitronate formation, k(cat)/k(m), decreased from an upper limiting value ...200919199786
transient responses during hyperosmotic shock in the filamentous fungus neurospora crassa.fungal cells maintain an internal hydrostatic pressure (turgor) of about 400-500 kpa. in the filamentous fungus neurospora crassa, the initial cellular responses to hyperosmotic treatment are loss of turgor, a decrease in relative hyphal volume per unit length (within 1 min) and cell growth arrest; all recover over a period of 10-60 min due to increased net ion uptake and glycerol production. the electrical responses to hyperosmotic treatment are a transient depolarization of the potential (with ...200919246761
carotenoids and carotenogenic genes in podospora anserina: engineering of the carotenoid composition extends the life span of the mycelium.carotenoids have been identified in the fungus podospora anserina and a parallel pathway to neurosporene and beta-carotene was established. three genes for the beta-carotene branch have been cloned and their function elucidated. they correspond to the al-1, al-2 and al-3 genes from neurospora crassa. they were individually and in combinations over-expressed in p. anserina in order to modify the carotenoid composition qualitatively and quantitatively. in the resulting transformants, carotenoid sy ...200919277665
the polarisome component spa-2 localizes at the apex of neurospora crassa and partially colocalizes with the spitzenkörper.in fungal hyphae multiple protein complexes assemble at sites of apical growth to maintain cell polarity and promote nucleation of actin. polarity allows the directional traffic of vesicles to the spitzenkörper (spk) prior to fusing with the plasma membrane to provide precursors and enzymes required for cell extension and nutrition. one of these complexes is the polarisome, which in saccharomyces cerevisiae contains spa2p, pea2p, bud6p/aip3p and bni1p. to investigate the localization and role of ...200919281855
re-characterisation of saccharomyces cerevisiae ach1p: fungal coa-transferases are involved in acetic acid detoxification.saccharomyces cerevisiae and neurospora crassa mutants defective in the so-called acetyl-coa hydrolases ach1p and acu-8, respectively, display a severe growth defect on acetate, which is most strongly pronounced under acidic conditions. acetyl-coa hydrolysis is an energy wasting process and therefore denoted as a biochemical conundrum. acetyl-coa hydrolases show high sequence identity to the coa-transferase coat from aspergillus nidulans. therefore, we extensively re-characterised the yeast enzy ...200919298859
the basidiomycetous mushroom lentinula edodes white collar-2 homolog phrb, a partner of putative blue-light photoreceptor phra, binds to a specific site in the promoter region of the l. edodes tyrosinase gene.we isolated a cdna homolog of neurospora crassa wc-2 from the basidiomycetous mushroom lentinula edodes and termed it phrb cdna. the deduced phrb (313 amino acid residues) contained a pas domain and a zinc-finger motif. random binding-site selection analysis of the phrb produced in escherichia coli revealed that it bound to a 7-bp sequence with the consensus sequence 5'gata/ttg/t/ac3'. electrophoretic mobility-shift assay showed that it also bound to the consensus sequence 5'gatattc3' in the pro ...200919344689
growth at high ph and sodium and potassium tolerance in media above the cytoplasmic ph depend on ena atpases in ustilago maydis.potassium and na(+) effluxes across the plasma membrane are crucial processes for the ionic homeostasis of cells. in fungal cells, these effluxes are mediated by cation/h(+) antiporters and ena atpases. we have cloned and studied the functions of the two ena atpases of ustilago maydis, u. maydis ena1 (umena1) and umena2. umena1 is a typical k(+) or na(+) efflux atpase whose function is indispensable for growth at ph 9.0 and for even modest na(+) or k(+) tolerances above ph 8.0. umena1 locates to ...200919363061
unwinding by local strand separation is critical for the function of dead-box proteins as rna chaperones.the dead-box proteins cyt-19 in neurospora crassa and mss116p in saccharomyces cerevisiae are broadly acting rna chaperones that function in mitochondria to stimulate group i and group ii intron splicing and to activate mrna translation. previous studies showed that the s. cerevisiae cytosolic/nuclear dead-box protein ded1p could stimulate group ii intron splicing in vitro. here, we show that ded1p complements mitochondrial translation and group i and group ii intron splicing defects in mss116de ...200919393667
the mitochondrial proteome database: mitop2.defining the mitochondrial proteome is a prerequisite for fully understanding the organelles function as well as mechanisms underlying mitochondrial pathology. the core functions of mitochondria include oxidative phosphorylation, amino acid metabolism, fatty acid oxidation, and ion homeostasis. in addition to these well-known functions, many crucial properties in cell signaling, cell differentiation and cell death are only now being elucidated, and with them the proteins involved. with the wealt ...200919426859
a phylogenetically conserved dna damage response resets the circadian clock.the mammalian circadian clock influences the timing of many biological processes such as the sleep/wake cycle, metabolism, and cell division. environmental cues such as light exposure can influence the timing of this system through the posttranslational modification of key components of the core molecular oscillator. we have previously shown that dna damage can reset the circadian clock in a time-of-day-dependent manner in the filamentous fungus neurospora crassa through the modulation of negati ...200919465696
properties of the kinesin-3 nckin3 motor domain and implications for neck function.members of the kinesin-3 family are microtubule motors involved in the transport of membranous cargo. nckin3 from the fungus neurospora crassa is dimeric but inactivates one of its motor heads to generate nonprocessive motility. to determine how one of the heads is inactivated, we investigated truncated monomeric constructs. none of the constructs generated processive single-molecule motility, and multimotor velocities depended linearly on the number of residues remaining in the neck. the kineti ...200919490122
investigation of regulatory factors required for alternative oxidase production in neurospora crassa.alternative oxidase (aox) has been found in a large number of filamentous fungi and yeasts with the notable exceptions of saccharomyces cerevisiae and schizosaccharomyces pombe. in virtually all of these fungi, aox is induced by stresses on the cell that compromise the efficiency of the standard mitochondrial electron transport chain. as aox is encoded in the nucleus and the signals that induce its expression originate in mitochondria, induction of the enzyme provides a classic example of retrog ...200919493307
mapping the interaction sites of aspergillus nidulans phytochrome fpha with the global regulator vea and the white collar protein lreb.aspergillus nidulans senses red and blue-light and employs a phytochrome and a neurospora crassa white collar (wc) homologous system for light perception and transmits this information into developmental decisions. under light conditions it undergoes asexual development and in the dark it develops sexually. the phytochrome fpha consists of a light sensory domain and a signal output domain, consisting of a histidine kinase and a response regulator domain. previously it was shown that the phytochr ...200918936976
the cyanide hydratase from neurospora crassa forms a helix which has a dimeric repeat.the fungal cyanide hydratases form a functionally specialized subset of the nitrilases which catalyze the hydrolysis of cyanide to formamide with high specificity. these hold great promise for the bioremediation of cyanide wastes. the low resolution (3.0 nm) three-dimensional reconstruction of negatively stained recombinant cyanide hydratase fibers from the saprophytic fungus neurospora crassa by iterative helical real space reconstruction reveals that enzyme fibers display left-handed d(1) s(5. ...200918946669
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
characterization of gpit-1 and gpit-2, two auxiliary components of the neurospora crassa gpi transamidase complex.the glycosylphosphatidylinositol (gpi) transamidase contains five known subunits and functions in the lumen of the er to produce gpi-anchored proteins. the transamidase cleaves proteins containing a gpi anchor attachment signal at their c terminus and generates an amide bond between the newly generated carboxyl terminus of the protein and a gpi anchor. we have identified and characterized gpit-1 and gpit-2, two of the transamidase subunits from neurospora crassa. gpit-1 and gpit-2 are homologs o ...200919927742
functional characterization of tzn1 and tzn2-zinc transporter genes in neurospora crassa.previous work from our laboratory involved the description of the neurospora metal transportome, which included seven hypothetical zinc transporters belonging to the zip family. the aim of the present study was to make a comparative functional evaluation of two hypothetical zinc transporters named tzn1 (ncu07621.3) and tzn2 (ncu11414.3). phenotypic analysis of tzn1 and tzn2 mutants and a double mutant (tzn1tzn2) revealed that the deletion of tzn1 causes aconidiation and a greater defect in growt ...200918989627
the group iii two-component histidine kinase of filamentous fungi is involved in the fungicidal activity of the bacterial polyketide ambruticin.we have shown that the plant pathogen alternaria brassicicola exhibited very high susceptibility to ambruticin vs4 and to a lesser extent to the phenylpyrrole fungicide fludioxonil. these compounds are both derived from natural bacterial metabolites with antifungal properties and are thought to exert their toxicity by interfering with osmoregulation in filamentous fungi. disruption of the osmosensor group iii histidine kinase gene abnik1 (for a. brassicola nik1) resulted in high levels of resist ...200919011080
neurospora crassa mat a-2 and mat a-3 proteins weakly interact in the yeast two-hybrid system and affect yeast growth.mating-type genes control the entry into the sexual cycle, mating identity and sexual development in fungi. the mat a-2 and mat a-3 genes, present in the mat a idiomorph of the filamentous fungus neurospora crassa, are required for post-fertilization functions but are not essential for mating identity. their putative roles as transcription factors are based on the similarity of mat a-2 with the podospora anserina smr1 gene and an hmg motif present in the mat a-3 gene. in this work the yeast two- ...200921637691
identification and biochemical characterization of molybdenum cofactor-binding proteins from arabidopsis thaliana.the molybdenum cofactor (moco) forms part of the catalytic center in all eukaryotic molybdenum enzymes and is synthesized in a highly conserved pathway. among eukaryotes, very little is known about the processes taking place subsequent to moco biosynthesis, i.e. moco transfer, allocation, and insertion into molybdenum enzymes. in the model plant arabidopsis thaliana, we identified a novel protein family consisting of nine members that after recombinant expression are able to bind moco with k(d) ...200920040598
illuminating solution responses of a lov domain protein with photocoupled small-angle x-ray scattering.the pas-lov domain is a signal-transducing component found in a large variety of proteins that is responsible for sensing different stimuli such as light, oxygen, and voltage. the lov protein vvd regulates blue light responses in the filamentous fungi neurospora crassa. using photocoupled, time-resolved small-angle x-ray scattering, we extract the solution protein structure in both dark-adapted and light-activated states. two distinct dark-adapted conformations are detected in the wild-type prot ...200919712683
regulation of glutamine synthesis by glycine and serine in neurospora crassa.the biosynthetic activities of the polypeptide subunits alpha and beta of glutamine synthetase (gs) were inhibited in vitro by glycine and serine. these amino acids inhibited the growth of a mutant strain with partial gs activity when grown on glutamate as the nitrogen source and also blocked the synthesis of the glutamine in vivo, thus demonstrating the inhibitory effect on gs activity in vivo. glycine and serine lowered the intracellular glutamine pool and regulated gs beta synthesis. a prefer ...20092867084
dna sequence duplications trigger gene inactivation in neurospora crassa.transforming sequences are faithfully replicated in vegetative cells of neurospora but are typically subject at high frequency to sequence alterations and methylation in the period between fertilization and nuclear fusion. previous work showed a correlation between the occurrence of these radical changes, referred to by the acronym rip, and the presence of sequence duplications resulting from the introduced dna. various possible causes for the rip process were investigated. introduction of a sin ...20092842795
gene organization and regulation in the qa (quinic acid) gene cluster of neurospora crassa. 20092931582
nucleotide sequence of pho-4+, encoding a phosphate-repressible phosphate permease of neurospora crassa.the nucleotide (nt) sequence of the neurospora crassa pho-4+ gene, which encodes a phosphate-repressible phosphate permease, has been determined. the gene specifies a protein of 590 amino acids (aa) and contains two introns. two rna transcripts of 3.3 and 2.4 kb have been identified, and transcription start points (tsp) and termination sites and/or processing sites have been located. the 3.3-kb message is initiated about 890 nt upstream from the tsp for the 2.4-kb transcript. a hydropathy profil ...20082531109
restricted distribution of the tad transposon in strains of neurospora.a simple colony blot procedure was used to screen 336 neurospora strains for the presence of the transposable element tad. these strains included the standard laboratory wild types, all of the available neurospora isolates collected from the ivory coast, and all wild-collected neurospora crassa isolates available from the fungal genetics stock center. tad was found only in the strain of origin from adiopodumé, ivory coast, where it is present in multiple copies. three other strains of african or ...20082546685
fast light-regulated genes of neurospora crassa.several physiological reactions including the sexual differentiation of the ascomycete neurospora crassa are triggered by blue light. mutants in the white-collar genes wc-1 and wc-2 are blind for all the blue light effects tested so far. we have previously shown that blue light induces some translatable mrnas at different times after beginning the illumination. here we report the cdna cloning of four genes that are induced by blue light. induction of these transcripts is temporally ordered (lag ...20082527354
light activation of the lov protein vivid generates a rapidly exchanging dimer.the fungal photoreceptor vivid (vvd) plays an important role in the adaptation of blue-light responses in neurospora crassa. vvd, an fad-binding lov (light, oxygen, voltage) protein, couples light-induced cysteinyl adduct formation at the flavin ring to conformational changes in the n-terminal cap (ncap) of the vvd pas domain. size-exclusion chromatography (sec), equilibrium ultracentrifugation, and static and dynamic light scattering show that these conformational changes generate a rapidly exc ...200818553928
stress-induced cell death is mediated by ceramide synthesis in neurospora crassa.the combined stresses of moderate heat shock (45 degrees c) and analog-induced glucose deprivation constitute a lethal stress for neurospora crassa. we found that this cell death requires fatty acid synthesis and the cofactor biotin. in the absence of the cofactor, the stressed cells are particularly sensitive to exogenous ceramide, which is lethal at low concentrations. when we extracted endogenous sphingolipids, we found that unique ceramides were induced (i) by the inhibitory glucose analog 2 ...200818952903
ripcal: a tool for alignment-based analysis of repeat-induced point mutations in fungal genomic sequences.repeat-induced point mutation (rip) is a fungal-specific genome defence mechanism that alters the sequences of repetitive dna, thereby inactivating coding genes. repeated dna sequences align between mating and meiosis and both sequences undergo c:g to t:a transitions. in most fungi these transitions preferentially affect cpa di-nucleotides thus altering the frequency of certain di-nucleotides in the affected sequences. the majority of previously published in silico analyses were limited to the c ...200819014496
atf-1 transcription factor regulates the expression of ccg-1 and cat-1 genes in response to fludioxonil under os-2 map kinase in neurospora crassa.the atf/creb family transcriptional factors are regulated by stress-activated map kinase in yeast. the disruptants of the atf-1 gene, which encodes an atf/creb family transcriptional factor, were isolated and characterized in neurospora crassa. the characteristic phenotypes in the os-2 map kinase strain, such as osmotic sensitivity and fludioxonil resistance, were not observed in the deltaatf-1 strain; however, like the os-2 strain, up-regulation of the catalase gene cat-1 and the clock-controll ...200818948219
genetic characterization of morphologically variant strains of paracoccidioides brasiliensis.molecular characterization of paracoccidioides brasiliensis variant strains that had been preserved under mineral oil for decades was carried out by random amplified polymorphic dna analysis (rapd). on p. brasiliensis variants in the transitional phase and strains with typical morphology, rapd produced reproducible polymorphic amplification products that differentiated them. a dendrogram based on the generated rapd patterns placed the 14 p. brasiliensis strains into five groups with similarity c ...200818545858
distinct structural elements dictate the specificity of the type iii pentaketide synthase from neurospora crassa.the fungal type iii polyketide synthase 2'-oxoalkylresorcylic acid synthase (oras) primes with a range of acyl-coenzyme a thioesters (c4-c20) and extends using malonyl-coenzyme a to produce pyrones, resorcinols, and resorcylic acids. to gain insight into this unusual substrate specificity and product profile, we have determined the crystal structures of oras to 1.75 a resolution, the phe-252-->gly site-directed mutant to 2.1 a resolution, and a binary complex of oras with eicosanoic acid to 2.0 ...200818940668
genome mining of cyanide-degrading nitrilases from filamentous fungi.a variety of fungal species are known to degrade cyanide through the action of cyanide hydratases, a specialized subset of nitrilases which hydrolyze cyanide to formamide. in this paper, we report on two previously unknown and uncharacterized cyanide hydratases from neurospora crassa and aspergillus nidulans. recombinant forms of four cyanide hydratases from n. crassa, a. nidulans, gibberella zeae, and gloeocercospora sorghi were prepared after their genes were cloned with n-terminal hexahistidi ...200818587571
dissecting colony development of neurospora crassa using mrna profiling and comparative genomics approaches.colony development, which includes hyphal extension, branching, anastomosis, and asexual sporulation, is a fundamental aspect of the life cycle of filamentous fungi; genetic mechanisms underlying these phenomena are poorly understood. we conducted transcriptional profiling during colony development of the model filamentous fungus neurospora crassa, using 70-mer oligonucleotide microarrays. relative mrna expression levels were determined for six sections of defined age excised from a 27-h-old n. ...200818676954
direct interaction between dna methyltransferase dim-2 and hp1 is required for dna methylation in neurospora crassa.dna methylation is involved in gene silencing and genomic stability in mammals, plants, and fungi. genetics studies of neurospora crassa have revealed that a dna methyltransferase (dim-2), a histone h3k9 methyltransferase (dim-5), and heterochromatin protein 1 (hp1) are required for dna methylation. we explored the interrelationships of these components of the methylation machinery. a yeast two-hybrid screen revealed that hp1 interacts with dim-2. we confirmed the interaction in vivo and demonst ...200818678653
the nonoxidative conversion of nitroethane to ethylnitronate in neurospora crassa 2-nitropropane dioxygenase is catalyzed by histidine 196.the deprotonation of nitroethane catalyzed by neurospora crassa 2-nitropropane dioxygenase was investigated by measuring the formation and release of ethylnitronate formed in turnover as a function of ph and through mutagenesis studies. progress curves for the enzymatic reaction obtained by following the increase in absorbance at 228 nm over time were visibly nonlinear, requiring a logarithmic approximation of the initial reaction rates for the determination of the kinetic parameters of the enzy ...200818690716
evolutionary history of the ancient cutinase family in five filamentous ascomycetes reveals differential gene duplications and losses and in magnaporthe grisea shows evidence of sub- and neo-functionalization.* the cuticle is the first barrier for fungi that parasitize plants systematically or opportunistically. here, the evolutionary history is reported of the multimembered cutinase families of the plant pathogenic ascomycetes magnaporthe grisea, fusarium graminearum and botrytis cinerea and the saprotrophic ascomycetes aspergillus nidulans and neurospora crassa. * molecular taxonomy of all fungal cutinases demonstrates a clear division into two ancient subfamilies. no evidence was found for lateral ...200818713314
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