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large-scale expressed sequence tag analysis for the chestnut blight fungus cryphonectria parasitica.cryphonectria parasitica is the causal fungal agent responsible for the chestnut blight disease. we report the generation of 14,263 expressed sequence tags representing 6318 unisequences for the fungus. functional annotation of these unisequences revealed different gene expression patterns for wild-type and hypovirus-infected cultures at the sporulation stage and allowed the reconstruction of key c. parasitica signal transduction pathways conserved in the sorbidaryomycetes. a list of homologous ...200818166491
structure of a tyrosyl-trna synthetase splicing factor bound to a group i intron rna.the 'rna world' hypothesis holds that during evolution the structural and enzymatic functions initially served by rna were assumed by proteins, leading to the latter's domination of biological catalysis. this progression can still be seen in modern biology, where ribozymes, such as the ribosome and rnase p, have evolved into protein-dependent rna catalysts ('rnpzymes'). similarly, group i introns use rna-catalysed splicing reactions, but many function as rnpzymes bound to proteins that stabilize ...200818172503
cloning and characterization of the blr2, the homologue of the blue-light regulator of neurospora crassa wc-2, in the phytopathogenic fungus bipolaris oryzae.bipolaris oryzae is a filamentous ascomycetous fungus that causes brown leaf spot disease in rice. we isolated and characterized blr2, a gene that encodes a putative blue-light regulator similar to neurospora crassa white collar-2 (wc-2). the deduced amino acid sequence of the blr2 showed significant homology to other fungal blue-light regulator proteins in the per-arnt-sim (pas) protein-protein interaction domain, nuclear localization signal, and gata zinc finger dna-binding domains. the blr2-s ...200818172720
functional characterization of fusarium verticillioides cpp1, a gene encoding a putative protein phosphatase 2a catalytic subunit.fusarium verticillioides produces the mycotoxin fumonisin b(1) (fb(1)) on maize kernels. in this study, we identified a putative protein phosphatase gene cpp1 in f. verticillioides, and investigated its role in fb(1) regulation. previous work has shown that cpp1 expression is elevated in an fb(1)-suppressing genetic background. thus, we hypothesized that cpp1 is negatively associated with fb(1) production. to test this hypothesis, we generated a cpp1 knockout mutant, pp179, and studied the effec ...200818174151
hydrolysis and fermentation of brewer's spent grain by neurospora crassa.in this study, the ethanol production by the mesophilic fungus neurospora crassa from bg was studied and optimized concerning the induction of lignocellulose degrading enzymes and the production phase as well. the production of cellulolytic and hemicellulolytic enzymes was studied under solid-state cultivation (ssc). ssc in a laboratory horizontal bioreactor using the optimized medium, ws and bg in the ratio 1:1 and initial moisture level 61.5%, allowed the large scale production of the multienz ...200818178432
protein kinases fpk1p and fpk2p are novel regulators of phospholipid asymmetry.type 4 p-type atpases (flippases) are implicated in the generation of phospholipid asymmetry in membranes by the inward translocation of phospholipids. in budding yeast, the drs2/dnf family members lem3p-dnf1p/dnf2p and cdc50p-drs2p are putative flippases that are localized, respectively, to the plasma membrane and endosomal/trans-golgi network (tgn) compartments. herein, we identified a protein kinase gene, fpk1, as a mutation that exhibited synthetic lethality with the cdc50delta mutation. the ...200818199685
the white collar protein wcoa of fusarium fujikuroi is not essential for photocarotenogenesis, but is involved in the regulation of secondary metabolism and conidiation.the fungal proteins of the white collar photoreceptor family, represented by wc-1 from neurospora crassa, mediate the control by light of different biochemical and developmental processes, such as carotenogenesis or sporulation. carotenoid biosynthesis is induced by light in the gibberellin-producing fungus fusarium fujikuroi. in an attempt to identify the photoreceptor for this response, we cloned the only wc-1-like gene present in the available fusarium genomes, that we called wcoa. the predic ...200818203635
characterization of mutations in alhk1 gene from alternaria longipes: implication of limited function of two-component histidine kinase on conferring dicarboximide resistance.four series (s, m, r, and w) of alternaria longipes isolates were obtained based on consecutive induction with dimethachlon (dim) and ultraviolet irradiation. these isolates were then characterized according to their tolerance to dim, sensitivity to osmotic stress, and phenotypic properties. all the induced dim-resistant isolates showed a higher osmosensitivity than the parental strains, and the last generation was more resistant than the first generation in the m, r, and w series. in addition, ...200818239410
genetic and molecular analysis of the temperature-sensitive mutant un-17 carrying a mutation in the gene encoding poly(a)-polymerase in neurospora crassa.the un-17 mutant was originally isolated as an irreparable temperature-sensitive (ts) mutant in neurospora crassa. early experiments showed that cells of this mutant immediately stopped growing and died when the temperature of the culture was shifted from a permissive temperature (25 degrees c) to non-permissive temperature (35 degrees c). this ts phenotype is suppressed by addition of cycloheximide or in some conditions of growth repression. even at the permissive temperature, it shows a female ...200718270435
functional and physical interaction of blue- and red-light sensors in aspergillus nidulans.light sensing is very important for organisms in all biological kingdoms to adapt to changing environmental conditions. it was discovered recently that plant-like phytochrome is involved in light sensing in the filamentous fungus aspergillus nidulans[1]. here, we show that phytochrome (fpha) is part of a protein complex containing lrea (wc-1) and lreb (wc-2) [2, 3], two central components of the neurospora crassa blue-light-sensing system. we found that fpha represses sexual development and myco ...200818291652
phenotype of a mechanosensitive channel mutant, mid-1, in a filamentous fungus, neurospora crassa.in the yeast saccharomyces cerevisiae, the mid1 (mating-induced death) gene encodes a stretch-activated channel which is required for successful mating; the mutant phenotype is rescued by elevated extracellular calcium. homologs of the mid1 gene are found in fungi that are morphologically complex compared to yeast, both basidiomycetes and ascomycetes. we explored the phenotype of a mid-1 knockout mutant in the filamentous ascomycete neurospora crassa. the mutant exhibits lower growth vigor than ...200818296620
neurospora crassa as a model organism for mitochondrial biogenesis.neurospora crassa has proven to be an excellent organism for studying various aspects of the biology of mitochondria by biochemical and genetic approaches. as n. crassa is an obligate aerobe and contains complex i, its mitochondria are more similar to mammalian mitochondria than those of yeast. the recent sequencing of the genome of n. crassa and a gene knockout project that is under way make the organism even more attractive. we describe some of the advantages of n. crassa as a model organism a ...200718314721
[homology-dependent inactivation of ltr retrotransposons in genomes of aspergillus fumigatus and a. nidulans].repeat-induced point mutation (rip) is the most intriguing among the known mechanisms of repeated sequences inactivation because of its ability to produce irreversible mutation of repeated dna. discovered for the first time in neurospora crassa, rip is characterized by c:g to t:a transitions in duplicated sequences. the mechanisms and distribution of rip are still purely investigated. mobile elements are a common target for the processes which lead to homology-dependent silencing because of thei ...200718318114
repeat induced point mutation in two asexual fungi, aspergillus niger and penicillium chrysogenum.repeat induced point mutation (rip) is a gene silencing mechanism present in fungal genomes. during rip, duplicated sequences are efficiently and irreversibly mutated by transitions from c:g to t:a. for the first time, we have identified traces of rip in transposable elements of aspergillus niger and penicillium chrysogenum, two biotechnologically relevant fungi. we found that rip in p. chrysogenum has affected a large set of sequences, which also contain other mutations. on the other hand, rip ...200818347798
a novel gene, phca from pseudomonas syringae induces programmed cell death in the filamentous fungus neurospora crassa.the phytopathogen pseudomonas syringae competes with other epiphytic organisms, such as filamentous fungi, for resources. here we characterize a gene in p. syringae pv. syringae b728a and p. syringae pv. tomato dc3000, termed phca, that has homology to a filamentous fungal gene called het-c. phca is conserved in many p. syringae strains, but is absent in one of the major clades, which includes the p. syringae pathovar phaseolicola. in the filamentous fungus neurospora crassa, het-c regulates a c ...200818363647
properties of family 79 beta-glucuronidases that hydrolyze beta-glucuronosyl and 4-o-methyl-beta-glucuronosyl residues of arabinogalactan-protein.the carbohydrate moieties of arabinogalactan-proteins (agps), which are mainly composed of gal, l-ara, glca, and 4-me-glca residues, are essential for the physiological functions of these proteoglycans in higher plants. for this study, we have identified two genes encoding family 79 beta-glucuronidases, designated anglcaase and ncglcaase, in aspergillus niger and neurospora crassa, respectively, based on the amino acid sequence of a native beta-glucuronidase purified from a commercial pectolytic ...200818377882
specificity of commercial anti-spectrin antibody in the study of fungal and oomycete spectrin: cross-reaction with proteins other than spectrin.spectrin was first described in erythrocytes where it forms a filamentous network in the cytoplasmic face of the plasma membrane and participates in the membrane's structural integrity in addition to controlling the lateral mobility of integral membrane proteins. in fungi, spectrin-like proteins have been described in the plasma membrane, concentrated mainly in the region of maximum apical expansion. this localization led to the idea of a spectrin based membrane skeleton in fungi participating i ...200818378170
genetic analysis of the neurospora crassa rad14 homolog mus-43 and the rad10 homolog mus-44 reveals that they belong to the mus-38 pathway of two nucleotide excision repair systems.saccharomyces cerevisiae rad14 and rad10 proteins are essential for nucleotide excision repair (ner). rad14 is a uv-damaged dna binding protein and rad10 is a structure-specific endonuclease that functions in a complex with rad1. in this study, we identified and characterized the rad14 and rad10 homolog genes in neurospora crassa, which we named mus-43 and mus-44, respectively. disruption of mus-43 and mus-44 conferred sensitivity to uv and 4-nitroquinoline 1-oxide, but not to methyl methanesulf ...200818379129
identification and evolution of fungal mitochondrial tyrosyl-trna synthetases with group i intron splicing activity.the bifunctional neurospora crassa mitochondrial tyrosyl-trna synthetase (cyt-18 protein) both aminoacylates mitochondrial trna(tyr) and acts as a structure-stabilizing splicing cofactor for group i introns. previous studies showed that cyt-18 has distinct trna(tyr) and group i intron-binding sites, with the latter formed by three small "insertions" in the nucleotide-binding fold and other structural adaptations compared with nonsplicing bacterial tyrosyl-trna synthetases. here, analysis of geno ...200818413600
biological rhythms workshop ia: molecular basis of rhythms generation.current circadian models are based on genetic, biochemical, and structural data that, when combined, provide a comprehensive picture of the molecular basis for rhythms generation. these models describe three basic elements-input pathways, oscillator, and output pathways-to which each molecular component is assigned. the lines between these elements are often blurred because some proteins function in more than one element of the circadian system. the end result of these molecular oscillations is ...200718419259
an f-actin-depleted zone is present at the hyphal tip of invasive hyphae of neurospora crassa.the distribution of filamentous actin (f-actin) in invasive and noninvasive hyphae of the ascomycete neurospora crassa was investigated. eighty six percent of noninvasive hyphae had f-actin in the tip region compared to only 9% of invasive hyphae. the remaining 91% of the invasive hyphae had no obvious tip high concentration of f-actin staining; instead they had an f-actin-depleted zone in this region, although some f-actin, possibly associated with the spitzenkörper, remained at the tip. the si ...200818421548
dynamin-like protein-dependent formation of woronin bodies in saccharomyces cerevisiae upon heterologous expression of a single protein.filamentous ascomycetes harbor woronin bodies and glyoxysomes, two types of microbodies, within one cell at the same time. the dominant protein of the neurospora crassa woronin body, hex1, forms a hexagonal core crystal via oligomerization and evidence has accumulated that woronin bodies bud off from glyoxysomes. we analyzed whether hex1 is sufficient to induce woronin body formation upon heterologous expression in saccharomyces cerevisiae, an organism devoid of this specialized organelle. in wi ...200818435762
dynamics of the preprotein translocation channel of the outer membrane of mitochondria.the protein translocase of the outer mitochondrial membrane (tom) serves as the main entry site for virtually all mitochondrial proteins. like many other protein translocases it also has an ion channel activity that can be used to study the dynamical properties of this supramolecular complex. we have purified tom core complex and tom40, the main pore forming subunit, from mitochondria of the filamentous fungus neurospora crassa and incorporated them into planar lipid bilayers. we then examined t ...200818456827
the genome sequence of the model ascomycete fungus podospora anserina.the dung-inhabiting ascomycete fungus podospora anserina is a model used to study various aspects of eukaryotic and fungal biology, such as ageing, prions and sexual development.200818460219
structural insights into biosynthesis of resorcinolic lipids by a type iii polyketide synthase in neurospora crassa.microbial type iii polyketide synthases (pkss) have revealed remarkable mechanistic as well as functional versatility. recently, a type iii pks homolog from azotobacter has been implicated in the biosynthesis of resorcinolic lipids, thus adding a new functional significance to this class of proteins. here, we report the structural and mutational investigations of a novel type iii pks protein from neurospora crassa involved in the biosynthesis of resorcinolic metabolites by utilizing long chain f ...200818462950
sexual and vegetative compatibility genes in the aspergilli.gene flow within populations can occur by sexual and/or parasexual means. analyses of experimental and in silico work are presented relevant to possible gene flow within the aspergilli. first, the discovery of mating-type (mat) genes within certain species of aspergillus is described. the implications for self-fertility, sexuality in supposedly asexual species and possible uses as phylogenetic markers are discussed. second, the results of data mining for heterokaryon incompatibility (het) and pr ...200718490952
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
mode of action and properties of xylanase and beta-xylosidase from neurospora crassa.extracellular beta-xylosidase (1,4-beta-d-xylan xylohydrolase, ec 3.2.1.37) from culture filtrates of neurospora crassa was purified to homogeneity by preparative isoelectric focusing followed by gel electrophoresis. the molecular weight of the purified xylosidase was 83,000 d and the k(m) on p-nitrophenyl-beta-d-xyloside was 0.047mm. the homogeneous xylanase (1,4-beta-d-xylan xylanohydrolase, ec 3.2.1.8) and beta-xylosidase showed differences in their mode of action towards xylooligosaccharides ...198618555300
the nuclear dbf2-related kinase cot1 and the mitogen-activated protein kinases mak1 and mak2 genetically interact to regulate filamentous growth, hyphal fusion and sexual development in neurospora crassa.ndr kinases, such as neurospora crassa cot1, are important for cell differentiation and polar morphogenesis, yet their input signals as well as their integration into a cellular signaling context are still elusive. here, we identify the cot-1 suppressor gul-4 as mak-2 and show that mutants of the gul-4/mak-2 mitogen-activated protein (map) kinase pathway suppress cot-1 phenotypes along with a concomitant reduction in protein kinase a (pka) activity. furthermore, mak-2 pathway defects are partial ...200818562669
nadph oxidases nox-1 and nox-2 require the regulatory subunit nor-1 to control cell differentiation and growth in neurospora crassa.we have proposed that reactive oxygen species (ros) play essential roles in cell differentiation. enzymes belonging to the nadph oxidase (nox) family produce superoxide in a regulated manner. we have identified three distinct nox subfamilies in the fungal kingdom and have shown that noxa is required for sexual cell differentiation in aspergillus nidulans. here we show that neurospora crassa nox-1 elimination results in complete female sterility, decreased asexual development, and reduction of hy ...200818567788
multiple gene disruptions by marker recycling with highly efficient gene-targeting background (deltaligd) in aspergillus oryzae.previously we reported that double disruption of the proteinase genes (tppa and pepe) improved heterologous protein production by aspergillus oryzae (jin et al. appl microbiol biotechnol 76:1059-1068, 2007). since a. oryzae has 134 protease genes, the number of auxotrophy in a single host is limited for multiple disruptions of many protease genes. in order to rapidly perform multiple gene disruptions in a. oryzae, we generated the marker recycling system in highly efficient gene-targeting backgr ...200818574559
a cytosine methyltransferase homologue is essential for sexual development in aspergillus nidulans.the genome defense processes rip (repeat-induced point mutation) in the filamentous fungus neurospora crassa, and mip (methylation induced premeiotically) in the fungus ascobolus immersus depend on proteins with dna methyltransferase (dmt) domains. nevertheless, these proteins, rid and masc1, respectively, have not been demonstrated to have dmt activity. we discovered a close homologue in aspergillus nidulans, a fungus thought to have no methylation and no genome defense system comparable to rip ...200818575630
the production and properties of a new xylose reductase from fungus. neurospora crassa.neurospora crassa xi was found to ferment xylose and glucose simultaneously. xylose was the appropriate inducer for the production of xylose reductase that had two isoenzymes designated as ei and eii. both ei and eii, which were purified by affinity chromatography, had nadph-dependent xylose reductase activities. eii also had nadh-dependent activity, and ei is the only xylose reductase found so far without any nadh-dependent activity. ei and eii had mws of 30 kda and 27 kda, and pis of 5.6 and 5 ...199818576008
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
mitop2: an integrative tool for the analysis of the mitochondrial proteome.mitochondria are crucial for normal cell metabolism and maintenance. mitochondrial dysfunction has been implicated in a spectrum of human diseases, ranging from rare monogenic to common multifactorial disorders. important for the understanding of organelle function is the assignment of its constituents, and although over 1,500 proteins are predicted to be involved in mammalian mitochondrial function, so far only about 900 are assigned to mitochondria with reasonable certainty. continuing efforts ...200818780189
the information highways of a biotechnological workhorse--signal transduction in hypocrea jecorina.the ascomycete hypocrea jecorina (anamorph trichoderma reesei) is one of the most prolific producers of biomass-degrading enzymes and frequently termed an industrial workhorse. to compete for nutrients in its habitat despite its shortcoming in certain degradative enzymes, efficient perception and interpretation of environmental signals is indispensable. a better understanding of these signals as well as their transmission machinery can provide sources for improvement of biotechnological processe ...200818803869
novel apocarotenoid intermediates in neurospora crassa mutants imply a new biosynthetic reaction sequence leading to neurosporaxanthin formation.neurosporaxanthin, beta-apo-4'-carotenoic acid (c35), represents the end-product of the carotenoid pathway in neurospora crassa. it is supposed to be synthesized in three steps catalyzed by sequential al-2, cao-2 and ylo-1 activities: (i) cyclization of 3,4-didehydrolycopene (c40); (ii) cleavage of torulene into beta-apo-4'-carotenal (c35); and finally (iii) oxidation of beta-apo-4'-carotenal. however, analyses of the ylo-1 mutant revealed the accumulation of intermediates other than beta-apo-4' ...200818812228
sulphur metabolism and cellulase gene expression are connected processes in the filamentous fungus hypocrea jecorina (anamorph trichoderma reesei).sulphur compounds like cysteine, methionine and s-adenosylmethionine are essential for the viability of most cells. thus many organisms have developed a complex regulatory circuit that governs the expression of enzymes involved in sulphur assimilation and metabolism. in the filamentous fungus hypocrea jecorina (anamorph trichoderma reesei) little is known about the participants in this circuit.200818842142
mitogen-activated protein kinase cascade required for regulation of development and secondary metabolism in neurospora crassa.mitogen-activated protein kinase (mapk) signaling cascades are composed of mapk kinase kinases (mapkkks), mapk kinases (mapkks), and mapks. in this study, we characterize components of a mapk cascade in neurospora crassa (mik-1, mapkkk; mek-1, mapkk; and mak-1, mapk) homologous to that controlling cell wall integrity in saccharomyces cerevisiae. growth of basal hyphae is significantly reduced in mik-1, mek-1, and mak-1 deletion mutants on solid medium. all three mutants formed short aerial hypha ...200818849472
mglig4, a homolog of neurospora crassa mus-53 (dna ligase iv), is involved in, but not essential for, non-homologous end-joining events in magnaporthe grisea.in many eukaryotic organisms, the non-homologous end-joining (nhej) system is a major pathway for the repair of dna double-strand breaks (dsbs). dna ligase iv is a component of the nhej system and is strictly required for the nhej system in saccharomyces cerevisiae and in neurospora crassa. to investigate the functions of dna ligase iv in magnaporthe grisea, we generated deletion mutants of mglig4, which encodes a homolog of n. crassa dna ligase iv. mutants (mglig4) showed no defects in asexual ...200818854221
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
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
a point mutation in the two-component histidine kinase bcos-1 gene confers dicarboximide resistance in field isolates of botrytis cinerea.abstract partial dna fragments of botrytis cinerea field isolates encoding the putative osmosensor histidine kinase gene (bcos1) were cloned by polymerase chain reaction amplification and the predicted amino acid sequences were compared between dicarboximide-sensitive and resistant field isolates. the predicted bcos1p is highly homologous to osmosensor histidine kinase os1p from neurospora crassa including the n-terminal six tandem repeats of approximately 90 amino acids. four dicarboximide-resi ...200218944142
the mycoparasite ampelomyces quisqualis expresses exga encoding an exo-beta-1,3-glucanase in culture and during mycoparasitism.abstract ampelomyces quisqualis, a mycoparasite of fungi causing powdery mildews, exhibited high levels of extracellular exo-beta-1,3-glucanase activity in culture compared with neurospora crassa and gliocladium roseum. a. quisqualis culture filtrates affected powdery mildew caused by sphaerotheca fusca in a manner indicative of cell wall degradation, as determined by microscopic examination. a gene encoding an exo-beta-1,3-glucanase in a. quisqualis, designated exga, was isolated and sequenced. ...199918944674
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
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
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
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
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
the fate of gene duplicates in the genomes of fungal pathogens.understanding how molecular changes underlie phenotypic variation within and between species is one of the main goals of evolutionary biology and comparative genetics. the recent proliferation of sequenced fungal genomes offers a unique opportunity to start elucidating the extreme phenotypic diversity in the kingdom fungi.1-4 we attempted to investigate the contribution of gene families to the evolutionary forces shaping the diversity of pathogenic lifestyles among the fungi.5 we studied a famil ...200819513258
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
structure and applications of a rhamnolipid surfactant produced in soybean oil waste.soybean oil soapstock was utilized as an alternative carbon source for the production of rhamnolipids by pseudomonas aeruginosa lbi strain. the chemical composition and properties of the rhamnolipid mixture obtained were determined to define its potential applications. the chemical characterization of the rhamnolipid has revealed the presence of ten different homologues. the monorhamnolipid rhac10c1) and the dirhamnolipid rha2c10c10 were the main components of the mixture that showed predominanc ...201019649781
expression of ribonuclease a and ribonuclease n1 in the filamentous fungus neurospora crassa.in this study, we investigated the ability of the fungus neurospora crassa to produce and secrete two ribonucleases: the heterologous bovine rnase a and the endogenous rnase n(1). a set of expression vectors was constructed, each consisting of an rnase a open reading frame under the control of a specific promoter and each with a specific terminator. n. crassa transformants were analyzed at the transcriptional and protein levels. irrespective of the promoter used, all transformants showed an rnas ...201019662399
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
purification and characterization of a low molecular weight endo-xylanase from mushroom termitomyces clypeatus.a low molecular weight endo-xylanase (ec 3.2.1.8) was purified from an edible mushroom termitomyces clypeatus grown in submerged medium with oat spelt xylan. xylanase was purified to apparent homogeneity by ammonium sulfate fractionation and gel filtration chromatography. its molecular weight was determined by gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 12 kda. the enzyme was found to be most active at 50 degrees c and ph 5.0, being most stab ...201019728166
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
camp signaling pathway controls glycogen metabolism in neurospora crassa by regulating the glycogen synthase gene expression and phosphorylation.the camp-pka signaling pathway plays an important role in many biological processes including glycogen metabolism. in this work we investigated its role in the neurospora crassa glycogen metabolism control using mutant strains affected in components of the pathway, the cr-1 strain deficient in adenylyl cyclase activity therefore has the pka pathway not active, and the mcb strain a temperature-sensitive mutant defective in the regulatory subunit of pka therefore is a strain with constitutively ac ...201019883780
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
aoso protein accumulates at the septal pore in response to various stresses in the filamentous fungus aspergillus oryzae.filamentous ascomycetes form hyphal networks that are compartmentalized by septa which have a perforated pore allowing the passage of cytoplasm and organelles between adjacent hyphal compartments. thus, the septal pore may play an important role in the organized growth of multicellular organisms. upon hyphal injury, the septal pore is plugged by a wound-healing organelle, known as the woronin body, to prevent excessive cytoplasmic leakage. however, the movement of proteins towards the septal por ...201019945422
the mitochondrial porin, vdac, has retained the ability to be assembled in the bacterial outer membrane.beta-barrel proteins are present in the outer membranes (oms) of gram-negative bacteria, mitochondria, and chloroplasts. their assembly requires a machinery of which the central component, called omp85 (bama) in bacteria and tob55 (sam50) in mitochondria, is evolutionarily conserved. an open question is whether the signals in beta-barrel om proteins required for assembly via this multicomponent machinery are also conserved. to address this question, we have expressed in escherichia coli the mito ...201019959601
the s helix mediates signal transmission as a hamp domain coiled-coil extension in the narx nitrate sensor from escherichia coli k-12.in the nitrate-responsive, homodimeric narx sensor, two cytoplasmic membrane alpha-helices delimit the periplasmic ligand-binding domain. the hamp domain, a four-helix parallel coiled-coil built from two alpha-helices (hd1 and hd2), immediately follows the second transmembrane helix. previous computational studies identified a likely coiled-coil-forming alpha-helix, the signaling helix (s helix), in a range of signaling proteins, including eucaryal receptor guanylyl cyclases, but its function re ...201019966007
the ndr kinase dbf-2 is involved in regulation of mitosis, conidial development, and glycogen metabolism in neurospora crassa.neurospora crassa dbf-2 encodes an ndr (nuclear dbf2-related) protein kinase, homologous to lats1, a core component of the hippo pathway. this pathway plays important roles in restraining cell proliferation and promoting apoptosis in differentiating cells. here, we demonstrate that dbf-2 is involved in three fundamental processes in a filamentous fungus: cell cycle regulation, glycogen biosynthesis, and conidiation. dbf-2 is predominantly localized to the nucleus, and most (approximately 60%) db ...201019966031
characterization of ctr family genes and the elucidation of their role in the life cycle of neurospora crassa.transcriptional analysis using qrt-pcr of 62 metal ion transporters during conidial germination of neurospora crassa showed a significant up regulation of a hypothetical copper transporter gene, tcu-1, that belongs to the ctr family. herein we characterised the ctr family genes (tcu-1, tcu-2 and tcu-3) and deciphered their role in various developmental phases of the n. crassa life cycle. cross complementation assays in copper uptake mutant of saccharomyces cerevisiae revealed that tcu-1, tcu-2 a ...201020034585
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