phosphate transporters from the higher plant arabidopsis thaliana.two cdnas (atpt1 and atpt2) encoding plant phosphate transporters have been isolated from a library prepared with mrna extracted from phosphate-starved arabidopsis thaliana roots, the encoded polypeptides are 78% identical to each other and show high degree of amino acid sequence similarity with high-affinity phosphate transporters of saccharomyces cerevisiae, neurospora crassa, and the mycorrhizal fungus glomus versiforme. the atpt1 and atpt2 polypeptides are integral membrane proteins predicte ...19968927627
blue light induced adp ribosylation of 38 and 56 kda proteins in the soluble fraction of mycelia of neurospora crassa.soluble fractions prepared from the mycelia of wild type (74-or23-1a) and band (bd) exhibited an increase in the rate of the adp ribosylation of a 38 kda protein from nicotinamide adenine [32p]dinucleotide ([32p]nad) in the presence of 10(-7) m riboflavin caused by blue light irradiation in vitro. the soluble fraction was mixed with a reaction mixture containing 5 microci [32p]nad at 0 degree c for 20 s and then it was irradiated with blue light (420 nm, 42 mumol m-2 s-1) for 12.5, 25, 50, 100, ...19968933725
cloning and sequencing of the lys1 gene encoding homocitrate synthase in the yeast yarrowia lipolytica.the alpha-aminoadipate pathway for the biosynthesis of lysine is present only in fungi and euglena. the first step in the pathway is the condensation of acetyl-coa and alpha-ketoglutarate into homocitrate, and this step is carried out by the enzyme homocitrate synthase (ec in spite of extensive genetic analysis, no mutation affecting this step has been isolated until now in model organisms such as saccharomyces cerevisiae or neurospora crassa, although identification of mutations affe ...19968948100
a mitochondrial signal peptide from neurospora crassa increases the permeability of isolated rat liver mitochondria.mitochondria that contain ca2+ can be induced by a variety of triggering agents and conditions to undergo a permeability transition (pt); the inner membrane becomes nonselectively permeable to small solutes. mastoparan, an amphipathic peptide from wasp venom, has recently been reported to induce this transition (pfeiffer et al., 1995, j. biol. chem. 270,4923). we have examined the effect on the permeability of isolated rat liver mitochondria of a second amphipathic peptide, the signal sequence o ...19968951036
the chsb gene of aspergillus nidulans is necessary for normal hyphal growth and development.the chsb gene from aspergillus nidulans encodes a class iii chitin synthase, an enzyme class found in filamentous fungi but not in yeast-like organisms. using a novel method, we isolated haploid segregants carrying a disrupted chsb allele from heterozygous diploid disruptants. the haploid disruptants grow as minute colonies that do not conidiate. hyphae from the disruptants have enlarged tips, a high degree of branching, and disorganized lateral walls. the mycelium is not deficient in chitin con ...19968953267
saccharomyces cerevisiae chorismate synthase has a flavin reductase activity.chorismate synthase (cs) catalyses the conversion of 5-enolpyruvylshikimate 3-phosphate (epsp) to form chorismate, which is the last common intermediate in the synthesis of the three aromatic amino acids phenylalanine, tyrosine and tryptophan. despite the overall redox-neutral reaction, catalysis has an absolute requirement for reduced flavin. in the fungus neurospora crassa, a flavin reductase (fr) activity able to generate reduced flavin mononucleotide in the presence of nadph is an intrinsic ...19968971708
cloning and functional analysis of a beta-tubulin gene from a benomyl resistant mutant of aspergillus parasiticus.a genomic dna library prepared from a benomyl resistant strain of aspergillus parasiticus was screened with a neurospora crassa beta-tubulin gene probe. a unique a. parasiticus genomic dna fragment, thought to carry a mutant beta-tubulin gene (benr), was isolated. two plasmids, pyt1 and pytpyrg, carrying the putative benr gene or benr plus a second selectable marker (pyrg), respectively, were used to transform a benomyl sensitive strain of a. parasiticus (cs10) to determine if benr conferred ben ...19968982061
functional promoter elements common to ribosomal protein and ribosomal rna genes in neurospora crassa.the promoter sequences of a cytoplasmic ribosomal protein gene (crp-2) of neurospora crassa were identified using promoter deletion and substitution mutants. a gene-targeting strategy was used to assay the mutants in vivo. the promoter architecture of crp-2 is complex and is more similar to that of ribosomal protein genes in mouse than in saccharomyces cerevisiae. six regions were identified as important for transcription. these included two elements, a cg repeat and a dde box, that are conserve ...19969003305
subunit vii of ubiquinol:cytochrome-c oxidoreductase from neurospora crassa is functional in yeast and has an n-terminal extension that is not essential for mitochondrial targeting.cdna clones encoding subunit vii of the neurospora crassa bc1 complex (ubiquinol:cytochrome-c oxidoreductase), which is homologous with subunit viii of the complex from yeast (encoded by qcr8), were identified on the basis of functional complementation of a yeast qcr8 deletion strain. the clones contain an open reading frame encoding a protein with a calculated molecular mass of 11.8 kda. the n-terminal eight residues of the amino acid sequence deduced from the cdna clones are absent from the ma ...19969003361
the aspergillus niger gcn4 homologue, cpca, is transcriptionally regulated and encodes an unusual leucine zipper.the general control transcriptional regulator gene cpca of aspergillus niger was cloned by complementation of a saccharomyces cerevisiae delta gcn4 mutant strain. the encoded protein conferred resistance to amino acid analogues when expressed in yeast. disruption of cpca in a. niger resulted in a strain which is sensitive towards 3-aminotriazole and fails to respond to amino acid starvation, cpca encodes a transcript of approximately 2400 nucleotides in length that includes a 5' leader region of ...19979004217
cloning and characterization of a neurospora crassa ribosomal protein gene, crps-7.a neurospora crassa ribosomal protein gene, crps-7, was isolated from a genomic dna library closely linked to the morphological gene ro-2. sequence analysis and a computerized database search revealed a high degree of homology to the xenopus laevis rps8 and rat rps7 gene, as well as to uncharacterized orfs from two yeast species. comparison with a nearly full-length cdna clone revealed two introns, one of which is in a conserved position shared with the xenopus gene. although a number of sequenc ...19979021131
serine/threonine protein phosphatases and a protein phosphatase 1 inhibitor from neurospora crassa.the major spontaneously active serine/threonine (ser/thr) protein phosphatase activities in n. crassa wild type (fgsc 424) were type-1 (pp1), type-2a (pp2a) and type-2c (pp2c). pp1 and pp2c predominantly dephosphorylated phosphorylase a and casein, respectively. pp2a acted on both substrates, but was two-fold more active against casein. pp1 activity was inhibited by protamine, heparin, okadaic acid (ic50 50 nm) and mammalian inhibitor-1 (ic50 2 nm). on the other hand. pp2a activity was inhibited ...19969033809
three-dimensional structure of nadh-dehydrogenase from neurospora crassa by electron microscopy and conical tilt reconstruction.nadh-dehydrogenase (complex i) is the first complex of the mitochondrial respiratory chain. it is an amphipatic molecule located in the inner mitochondrial membrane and is composed of at least 35 unique subunits encoded by both mitochondrial and nuclear dna. the whole complex was isolated in detergent from the fungus neurospora crassa. it is very stable in its isolated form and was analysed as such by electron microscopy. its mass, determined by dark-field scanning electron microscopy was estima ...19979034360
analysis of frequency (frq) clock gene homologs: evidence for a helix-turn-helix transcription factor.the frq gene plays a key role in the organization of the neurospora crassa circadian clock. our previous analysis of a c-terminal fragment of the putative frq protein suggested it is a nuclear transcription factor but did not identify a known dna-binding domain. if the hypothesis is correct that frq is a transcription factor, sequences consistent with this function should be conserved in distantly related species. to investigate, we have cloned frq homologs from other filamentous fungi including ...19979037100
cross-pathway and pathway-specific control of amino acid biosynthesis in magnaporthe griseathe gene encoding the small subunit of the arginine-specific carbamoyl phosphate synthetase, arg2, of magnaporthe grisea was characterized to examine the basic regulation of biosynthetic genes in this plant pathogen. the transcript of the arg2 gene contains an upstream open reading frame (uorf) that is similar to uorfs found in the homologous genes of neurospora crassa (arg-2) and saccharomyces cerevisiae (cpa1), suggesting that the m. grisea gene is translationally regulated by a mechanism that ...19979073479
efficient cloning of ascomycete mating type genes by pcr amplification of the conserved mat hmg boxcloning of mating type (mat) genes from ascomycetes has been hampered by low conservation among them. one of the pair of mat genes, represented by mat-2 of cochliobolus heterostrophus (a loculoascomycete) and mt a of neurospora crassa (a pyrenomycete), encodes a protein with a conserved dna binding motif called the high mobility group (hmg) box. pcr with primer pairs corresponding to the borders of the c. heterostrophus and the n. crassa hmg boxes generated an approximately 0.3-kb product from g ...19979073486
effect of nonylphenol on growth of neurospora crassa and candida albicans.the effects of the nonionic surfactant nonylphenol on the growth and morphologies of the filamentous fungus neurospora crassa and the diploid yeast candida albicans have been examined. nonylphenol inhibited respiration and growth of n. crassa, effecting a 10-fold decrease in organism yield at 25 microm. severe morphological defects were also induced: cell shape was abnormal and apical dominance was lost. nonylphenol monoethoxylate (the parent compound of nonylphenol) was a less potent growth inh ...19979097428
genetic regulation of nitrogen metabolism in the the fungi, nitrogen metabolism is controlled by a complex genetic regulatory circuit which ensures the preferential use of primary nitrogen sources and also confers the ability to use many different secondary nitrogen sources when appropriate. most structural genes encoding nitrogen catabolic enzymes are subject to nitrogen catabolite repression, mediated by positive-acting transcription factors of the gata family of proteins. however, certain gata family members, such as the yeast dal80 fact ...19979106362
phospholipid composition of highly purified mitochondrial outer membranes of rat liver and neurospora crassa. is cardiolipin present in the mitochondrial outer membrane?isolated mitochondrial outer membrane vesicles (omv) are a suitable system for studying various functions of the mitochondrial outer membrane. for studies on mitochondrial lipid import as well as for studies on the role of lipids in processes occurring in the outer membrane, knowledge of the phospholipid composition of the outer membrane is indispensable. recently, a mild subfractionation procedure was described for the isolation of highly purified omv from mitochondria of neurospora crassa (may ...19979106488
cross-pathway and pathway-specific control of amino acid biosynthesis in magnaporthe grisea.the gene encoding the small subunit of the arginine-specific carbamoyl phosphate synthetase, arg2, of magnaporthe grisea was characterized to examine the basic regulation of biosynthetic genes in this plant pathogen. the transcript of the arg2 gene contains an upstream open reading frame (uorf) that is similar to uorfs found in the homologous genes of neurospora crassa (arg-2) and saccharomyces cerevisiae (cpa1), suggesting that the m. grisea gene is translationally regulated by a mechanism that ...19979126616
efficient cloning of ascomycete mating type genes by pcr amplification of the conserved mat hmg box.cloning of mating type (mat) genes from ascomycetes has been hampered by low conservation among them. one of the pair of mat genes, represented by mat-2 of cochliobolus heterostrophus (a loculoascomycete) and mt a of neurospora crassa (a pyrenomycete), encodes a protein with a conserved dna binding motif called the high mobility group (hmg) box. pcr with primer pairs corresponding to the borders of the c. heterostrophus and the n. crassa hmg boxes generated an approximately 0.3-kb product from g ...19979126621
quantitative and qualitative comparisons of spontaneous and radiation-induced specific-locus mutation in the ad-3 region of heterokaryon 12 of neurospora crassa.the data from forward-mutation experiments to obtain specific-locus mutations at two closely linked loci in the adenine-3 (ad-3) region of heterokaryon 12 (h-12) of neurospora crassa have been used to determine the relative frequencies and mutational spectra of ad-3 mutants occurring spontaneously and those induced by 7 different radiation treatments. previous studies have demonstrated that specific-locus mutants at these two loci result from 5 major genotypic classes, namely two classes of gene ...19979129678
quantitative and qualitative comparison of spontaneous and chemical-induced specific-locus mutation in the ad-3 region of heterokaryon 12 of neurospora crassa.the data from forward-mutation experiments to obtain specific-locus mutations at two closely linked loci in the adenine-3 (ad-3) region of heterokaryon 12 (h-12) of neurospora crassa have been tabulated to determine the relative frequencies and mutational spectra of ad-3 mutants occurring spontaneously and those induced by 22 different chemical treatments. previous studies have demonstrated that specific-locus mutations at these two loci result from 5 major genotypic classes, namely two classes ...19979129679
cloning of cutinase transcription factor 1, a transactivating protein containing cys6zn2 binuclear cluster dna-binding motif.hydroxy fatty acids from plant cutin were shown previously to induce the expression of the cutinase gene via a palindromic sequence located at -159 base pairs of the cutinase gene in fusarium solani f. sp. pisi (nectria hematococca mating type vi). of the two overlapping palindromes in this sequence, palindrome 2 was found to be essential for the inducibility of cutinase by hydroxy fatty acids. screening of a phage expression library with the concatenated palindrome 2 as probe detected a distinc ...19979139694
the osmotic-1 locus of neurospora crassa encodes a putative histidine kinase similar to osmosensors of bacteria and yeast.osmotically sensitive mutants of neurospora crassa are unable to grow on medium supplemented with 4% nacl, have altered morphologies and cell-wall compositions, and are resistant to dicarboximide fungicides. osmotic-1 (os-1) mutants have a unique characteristic of forming protoplasts that grow and divide in specialized liquid medium, suggesting that the os-1+ gene product is important for cell-wall assembly. a cosmid containing the os-1+ locus of n. crassa, isolated from a genomic cosmid library ...19979142740
identification of putative phosphoinositide-specific phospholipase c genes in filamentous fungi.five putative phosphoinositide-specific phospholipases c (plc) genes were identified in three species of filamentous fungi. using polymerase chain reaction with degenerate oligonucleotide primers, gene fragments encoding amino acid sequences homologous to plcs of mammals and other organisms were amplified: one sequence from botryotinia fuckeliana, one from aspergillus nidulans, and three from neurospora crassa. the molecular cloning and sequencing of a putative plc gene (bcplc1) from b. fuckelia ...19979163731
purification and characterization of an antifungal pr-5 protein from pumpkin leaves.a 28-kda antifungal pr-5 protein (pltp) was purified from pumpkin leaves to homogeneity by using ammonium sulfate fractionation, a regenerated chitin column, and reversed-phase column chromatographies on butyl-toyopearl and hplc c18 columns. analysis of 14 n-terminal amino acid sequences of pltp shows 100% sequence identity to those of two pr-5 proteins, np24 from tomatoes and ap24 from tobacco. the identical sequence also exhibited high amino acid sequence homology to that of an osmotin-like pr ...19979163735
cloning of an e. coli reca and yeast rad51 homolog, rada, an allele of the uvsc in aspergillus nidulans and its mutator e. coli reca and yeast rad51 homolog from aspergillus nidulans, rada, has been cloned by screening genomic and cdna libraries with a pcr-amplified probe. this probe was generated using primers carrying the conserved sequences of eukaryotic reca homologs. the deduced amino acid sequence revealed two conserved walker-a and -b type nucleotide-binding domains and exhibited 88%, 60%, and 53% identity with mei-3 of neurospora crassa, rhp51+ of schizosaccharomyces pombe, and rad51 of saccharomyces c ...19979163746
molecular cloning and expression studies of two divergent alpha-tubulin genes in neurospora crassa.three alpha-tubulin isoforms were previously detected in neurospora crassa. we have cloned and analysed two alpha-tubulin cdnas, tub alpha a and tub alpha b that encode polypeptides of 453 and 451 amino acids, respectively. the encoded amino acids exhibit an unusual divergence of 35%. this is the highest divergence ever observed between alpha-tubulins from the same species. the expression of the two genes is developmentally regulated. we did not detect any transcription of the tub alpha a gene i ...19979163903
cloning and characterization of a cdna encoding a bacteriophage-type rna polymerase from the higher plant chenopodium album.we have cloned a full-length cdna from the higher plant chenopodium album coding for a single subunit bacteriophage-type rna polymerase. the cdna isolated from an actively growing cell suspension culture recognized a 3.8 kb transcript on northern blots. the open reading frame comprises 987 amino acids with a predicted molecular mass of 112 kda. a comparison of the protein sequence with those of the two known fungal mitochondrial rna polymerases, from saccharomyces cerevisiae and neurospora crass ...19979171081
expression of herpes virus thymidine kinase in neurospora crassa.the expression of thymidine kinase in fungi, which normally lack this enzyme, will greatly aid the study of dna metabolism and provide useful drug-sensitive phenotypes. the herpes simplex virus type-1 thymidine kinase gene ( tk ) was expressed in neurospora crassa. tk was expressed as a fusion to n.crassa arg-2 regulatory sequences and as a hygromycin phosphotransferase-thymidine kinase fusion gene under the control of cytomegalovirus and sv40 sequences. only strains containing tk showed thymidi ...19979171090
rco-3, a gene involved in glucose transport and conidiation in neurospora crassa.macroconidiation in neurospora crassa is influenced by a number of environmental cues, including the nutritional status of the growing organism. conidia formation is normally observed when the fungus is exposed to air. however, carbon limitation can induce conidiation in mycclia submerged in an aerated liquid medium. a mutant was previously isolated that could conidiate in submerged culture without imposing nutrient limitation and the gene responsible for this phenotype (rco-3) has now been clon ...19979178001
a putative rhamnogalacturonase required for sexual development of neurospora previous work, the asd-i (ascus development) gene of the filamentous fingus neurospora crassa was identified as a gene expressed preferentially during the sexual cycle and shown to be essential for normal sexual development. the asd-i gene has been sequenced and further characterized. it contains two introns, the first of which is in-frame and inefficiently or differentially spliced. the predicted asd-i protein has extensive homology with rhamnogalacturonase b of aspergillus aculeatus, which ...19979178004
histidine kinases in signal transduction pathways of eukaryotes.autophosphorylating histidine kinases are an ancient conserved family of enzymes that are found in eubacteria, archaebacteria and eukaryotes. they are activated by a wide range of extracellular signals and transfer phosphate moieties to aspartates found in response regulators. recent studies have shown that such two-component signal transduction pathways mediate osmoregulation in saccharomyces cerevisiae, dictyostelium discoideum and neurospora crassa. moreover, they play pivotal roles in respon ...19979191038
adaptation of proteases and carbohydrates of saprophytic, phytopathogenic and entomopathogenic fungi to the requirements of their ecological niches.the abilities of isolates of saprophytes (neurospora crassa, aspergillus nidulans), an opportunistic human pathogen (aspergillus fumigatus), an opportunistic insect pathogen (aspergillus flavus), plant pathogens (verticillium albo-atrum, verticillium dahliae, nectria haematococca), a mushroom pathogen (verticillium fungicola) and entomopathogens (verticillium lecanii, beauveria bassiana, metarhizium anisopliae) to utilize plant cell walls and insect cuticle components in different nutrient media ...19979202474
isolation of developmentally regulated genes from the edible mushroom agaricus bisporus.from a cdna library, constructed from mushroom primordia, nine cdnas were isolated which were either induced or specifically expressed during fruit body development and maturation of the basidiomycete agaricus bisporus. these cdnas varied in size from 372 to 1019 bp and hybridized to transcripts of 400-1600 nt. four of the cdnas were only expressed in the generative phase of the life cycle while the other five cdnas were strongly induced but had low steady-state mrna levels in vegetatively grown ...19979202475
a kinesin-like mechanoenzyme from the zygomycete syncephalastrum racemosum shares biochemical similarities with conventional kinesin from neurospora crassa.the kinesin superfamily consists of mechanoenzymes that convert chemical energy, stored in nucleoside triphosphates, into movement along microtubules. the founding member of this protein superfamily, the so-called conventional kinesin, was only known from animal sources until the recent description of kinesin from the filamentous fungus neurospora crassa (g. steinberg, m. schliwa, mol. biol. cell 6, 1605-1618 (1995)). to determine whether similar motors with comparable features are common in oth ...19979208225
identification of a motor protein required for filamentous growth in ustilago maydis.the phytopathogenic fungus ustilago maydis exists in two stages, the yeast-like haploid form and the filamentous dikaryon. both pathogenicity and dimorphism are genetically controlled by two mating-type loci, with only the filamentous stage being pathogenic on corn. we have identified two genes (kin1 and kin2) encoding motor proteins of the kinesin family. kin1 is most similar to the human cenp-e gene product, while kin2 is most closely related to the conventional kinesin nkin of neurospora cras ...19979218789
footprinting of the spinach nitrite reductase gene promoter reveals the preservation of nitrate regulatory elements between fungi and higher plants.nitrite reductase (nir) is the second enzyme in the nitrate assimilatory pathway reducing nitrite to ammonium. the expression of the nir gene is induced upon the addition of nitrate. in an earlier study, a 130 bp upstream region of the spinach nir gene promoter, located between -330 to - 200, was shown to be necessary for nitrate induction of beta-glucuronidase (gus) expression in tissue-specific manner in transgenic tobacco plant [28]. to further delineate the cis-acting elements involved in ni ...19979225857
the geranylgeranyl diphosphate synthase gene of gibberella fujikuroi: isolation and expression.the rice pathogen, gibberella fujikuroi, produces large amounts of gibberellins, a group of natural plant hormones, which induce the superelongation (bakanae) disease of rice. gibberellins are diterpenoid compounds which are synthesized via the isoprenoid pathway. here we report the isolation and molecular characterization of the geranylgeranyl diphosphate synthase (ggs) gene from g. fujikuroi. geranylgeranyl diphosphate synthase is a key enzyme in isoprenoid biosynthesis. southern blot analysis ...19979230902
gene targeting is locus dependent in the filamentous fungus aspergillus nidulans.the effect of altering the conditions of transformation on the efficiency of gene targeting in filamentous fungi was studied using aspergillus nidulans as a model organism. the niad and amds genes of a. nidulans, which are both involved in nitrogen source utilisation, were selected as target loci. homologous recombination of transforming dna at these loci resulted in niad and amds mutants with an impaired ability to utilise nitrate or acetamide as the sole nitrogen source, respectively. vectors ...19979236780
monoclonal antibodies as probes for fungal wall structure during morphogenesis.three monoclonal antibodies (mabs), s4d1, s3b3 and s1e5, were produced from hybridoma cell lines raised from mice immunized with hyphal walls of neurospora crassa and one (pax-1) from mice immunized with hyphal walls of paxillus involutus. in immunofluorescence studies, the three n. crassa mabs recognized epitopes with different patterns of distribution at the hyphal surface of n. crassa. s4d1 recognized an epitope which was present on the surface of both conidia and hyphae; s3b3 recognized an e ...19979245814
isolation and characterisation of a novel stress-inducible pdi-family gene from aspergillus niger.current strategies to improve the secretion of heterologous proteins in aspergillus niger include the manipulation of chaperones and foldases specific to the endoplasmic reticulum (er). a family of er-specific proteins which share active-site homology wit protein disulfide isomerase (pdi) has been identified from other systems, many of which are inducible by agents which cause malfolding of proteins in the er. here we report identification of tiga from aspergillus niger and erp38 from neurospora ...19979256071
isolation and characterization of the krev-1 gene, a novel member of ras superfamily in neurospora crassa: involvement in sexual cycle progression.genes belonging to the ras superfamily encode low-molecular-weight gtp/gdp-binding proteins that are highly conserved in wide variety of organisms. we used the polymerase chain reaction (pcr) to isolate a novel member of the ras superfamily from the filamentous fungus neurospora crassa and obtained a mammalian krev-1 homolog. we named the gene krev-1 and analyzed its structure and function. the krev-1 gene encodes a polypeptide of 225 amino acids, which is nearly 60% homologous to the mammalian ...19979267439
genetic regulation of the qa gene cluster of neurospora crassa: induction of qa messenger ribonucleic acid and dependency on qa-1 in vitro protein-synthesizing system (rabbit reticulocyte) was programmed with total polyadenylated messenger ribonucleic acid from wild type and various mutants in the qa gene cluster of neurospora crassa. the products of two of the qa genes, quinate dehydrogenase (qa-3+) and dehydroshikimate dehydratase (qa-4+), were identified by specific immunoprecipitation and sodium dodecyl sulfate-slab gel electrophoresis. the results indicated that for both genes induction of a specific enzyme activit ...19819279395
reversal in the direction of movement of a molecular motor.kinesin and non-claret disjunctional (ncd) are molecular motors of the kinesin superfamily that move in opposite directions along microtubules. the molecular basis underlying the direction of movement is unclear, although it is thought to be an intrinsic property of the motor domain, a conserved region about 330 amino acids in length. the motor domain is found at the amino terminus in conventional kinesins, but at the carboxy terminus in ncd. here we report on a chimaera composed of the motor do ...19979288974
cloning heterologous genes: problems and approaches.model fungi such as neurospora crassa, aspergillus niger, and saccharomyces cerevisiae have provided a wealth of genetic information and are currently the object of cooperative genome sequencing projects. many agriculturally and medically economic important pathogenic fungi, however, are less well characterized, which makes it difficult to study their genes and gene products. gene sequences from model fungi offer a unique opportunity to clone cognate genes from pathogenic counterparts. in this r ...19979290241
contact-sensing by hyphae of dermatophytic and saprophytic or thigmotropism is the directional growth response of cells in relation to topographical guidance cues. thigmotropism is thought to play a major role in the location of infectable sites on plants by phytopathogenic fungi and has recently been shown to be a property of hyphae in the human pathogenic fungus candida albicans. here we show that hyphae of the dermatophytes epidermophyton floccosum, microsporum canis and trichophyton mentagrophytes reorientate their direction of growt ...19979292428
expression of a plant protein by neurospora crassa.heterologous expression of plant genes may serve as an important alternative for producing plant proteins. we have investigated the ability of the fungus neurospora crassa to secrete zeamatin, a protein produced by zea mays. zeamatin was induced after being fused to glucoamylase, an extracellular hydrolase produced by n. crassa. glucoamylase induction and other culture parameters were monitored in untransformed n. crassa grown in shaken liquid culture. a dna plasmid, pgez, was constructed by ins ...19979292999
the ornithine decarboxylase gene from candida albicans. sequence analysis and expression during dimorphism.the gene (caodc) coding for ornithine decarboxylase, a key enzyme in polyamine biosynthesis, was cloned from candida albicans by pcr and characterized. the deduced protein contains 470 amino acids together with the substrate- and co-factor-binding sequences which define the ornithine decarboxylases of eukaryotic species. it shows a high homology with other odcs, mainly those from saccharomyces cerevisiae and neurospora crassa. a putative pest sequence, which correlates quite well with those desc ...19979294258
fungal phosphate transporter serves as a receptor backbone for gibbon ape leukemia virus.pit1, the receptor for gibbon ape leukemia virus (galv), is proposed to be an integral membrane protein with five extracellular loops. chimeras made between pit1 homologs differing in permissivity for infection and between pit1 and the related protein pit2 have shown that the fourth extracellular loop plays a critical role in infection. however, further elucidation of the roles of the extracellular loops in infection is hampered by the high level of sequence similarity among these proteins. the ...19979311843
mating-type genes from the homothallic fungus sordaria macrospora are functionally expressed in a heterothallic ascomycete.homokaryons from the homothallic ascomycte sordaria macrospora are able to enter the sexual pathway and to form fertile fruiting bodies. to analyze the molecular basis of homothallism and to elucidate the role of mating-products during fruiting body development, we cloned and sequenced the entire s. macrospora mating-type locus. comparison of the sordaria mating-type locus with mating-type idiomorphs from the heterothallic ascomycetes neurospora crassa and podospora anserina revealed that sequen ...19979335594
the neurospora crassa cya-5 nuclear gene encodes a protein with a region of homology to the saccharomyces cerevisiae pet309 protein and is required in a post-transcriptional step for the expression of the mitochondrially encoded coxi protein.the cya-5 nuclear mutant of neurosopora crassa was previously shown to be deficient in cytochrome aa3, cytochrome c oxidase activity, and the immunologically detectable coxi protein. we have now demonstrated that the mitochondria of this mutant contain mrna for the coxi protein and that coxi cannot be detected during pulse-chase labeling experiments of mitochondrial translation products. cloning and analysis of the cya-5 gene reveal a long open reading frame capable of encoding a 1136 amino-acid ...19979342407
molecular genetics of sulfur assimilation in filamentous fungi and yeast.the filamentous fungi aspergillus nidulans and neurospora crassa and the yeast saccharomyces cerevisiae each possess a global regulatory circuit that controls the expression of permeases and enzymes that function both in the acquisition of sulfur from the environment and in its assimilation. control of the structural genes that specify an array of enzymes that catalyze reactions of sulfur metabolism occurs at the transcriptional level and involves both positive-acting and negative-acting regulat ...19979343344
mutations in genes encoding the mitochondrial outer membrane proteins tom70 and mdm10 of podospora anserina modify the spectrum of mitochondrial dna rearrangements associated with cellular death.tom70 and mdm10 are mitochondrial outer membrane proteins. tom70 is implicated in the import of proteins from the cytosol into the mitochondria in saccharomyces cerevisiae and neurospora crassa. mdm10 is involved in the morphology and distribution of mitochondria in s. cerevisiae. here we report on the characterization of the genes encoding these proteins in the filamentous fungus podospora anserina. the two genes were previously genetically identified through a systematic search for nuclear sup ...19979343397
impairment of calcineurin function in neurospora crassa reveals its essential role in hyphal growth, morphology and maintenance of the apical ca2+ gradient.the function of neurospora crassa calcineurin was investigated in n. crassa strains transformed with a construct that provides for the inducible expression of antisense rna for the catalytic subunit of calcineurin (cna-1). induction of antisense rna expression was associated with reduced levels of cna-1 mrna and of immunodetectable cna1 protein and decreased calcineurin enzyme activity, indicating that a conditional reduction of the target function had been achieved in antisense transformants wi ...19979349701
enzymatic properties and effect of ionic strength on periplasmic nitrate reductase (nap) from desulfovibrio desulfuricans atcc 27774.some sulfate reducing bacteria can induce nitrate reductase when grown on nitrate containing media being involved in dissimilatory reduction of nitrate, an important step of the nitrogen cycle. previously, it was reported the purification of the first soluble nitrate reductase from a sulfate-reducing bacteria desulfovibrio desulfuricans atcc 27774 (s.a. bursakov, m.-y. liu, w.j. payne, j. legall, i. moura, and j.j.g. moura (1995) anaerobe 1, 55-60). the present work provides further information ...19979367852
characterization of the mitochondrial cytochrome b gene from venturia inaequalis.a new class of agricultural fungicides derived from the group of antifungal strobilurins acts as specific respiration inhibitors by binding to mitochondrial cytochrome b. the cytochrome b gene was cloned and sequenced from the mitochondrial genome of venturia inaequalis, the causal agent of apple scab. the gene was 10.65 kbp in size and contained seven exons and six introns. the exons encoded a protein of 393 amino acids. comparison of the deduced amino-acid sequence with cytochrome b proteins f ...19979371888
g protein alpha subunit genes control growth, development, and pathogenicity of magnaporthe grisea.three g protein alpha subunit genes have been cloned and characterized from magnaporthe grisea: maga is very similar to cpg-2 of cryphonectria parasitica; magb is virtually identical to cpg-1 of cryphonectria parasitica, to gna1 of neurospora crassa, and to fada of emericella nidulans; and magc is most similar to gna2 of neurospora crassa. homologous recombination resulting in targeted deletion of maga had no effect on vegetative growth, conidiation, or appressorium formation. deletion of magc r ...19979390422
genetic and molecular characterization of neurospora crassa mus-23: a gene involved in recombinational repair.a newly isolated mutant, mus-23, of neurospora crassa was found to be highly sensitive to a wide variety of mutagens, including uv light, methyl methanesulfonate, 4-nitroquinoline 1-oxide, n-methyl-n'-nitro-n-nitrosoguanidine and tert-butyl hydroperoxide. this mutant was originally isolated as a mutant that could not grow on medium containing histidine. meiosis and sporulation were defective in homozygous crosses between mus-23 haploids. the mus-23 gene is located on the right arm of lgii, betwe ...19979393441
the production of gamma-linolenic acid by selected members of the dikaryomycota grown on different carbon this study, seven fungal strains, representing different phylogenetic groups within the dikaryomycota, were tested for the presence of gamma-linolenic acid [18:3(omega 6)], when grown in synthetic liquid media devoid of fatty acids, on a series of 40 different carbon sources. the fungal strains represented the species dipodascopsis uninucleata, eurotium rubrum, galactomyces geotrichum, neurospora crassa, saccharomyces cerevisiae, spongipellis unicolor and talaromyces flavus. cultures were per ...19979403104
arabidopsis nph1: a protein kinase with a putative redox-sensing domain.the nph1 (nonphototropic hypocotyl 1) gene encodes an essential component acting very early in the signal-transduction chain for phototropism. arabidopsis nph1 contains a serine-threonine kinase domain and lov1 and lov2 repeats that share similarity (36 to 56 percent) with halobacterium salinarium bat, azotobacter vinelandii nifl, neurospora crassa white collar-1, escherichia coli aer, and the eag family of potassium-channel proteins from drosophila and mammals. sequence similarity with a known ...19979405347
purification and characterization of the catalytic subunit of protein phosphatase 1 from neurospora crassa.protein phosphorylation is a universal regulatory mechanism in eukaryotic cells. the phosphorylation state of proteins is affected by the antagonistic activities of protein kinases and phosphatases. protein phosphatases (pps) can be classified as serine/threonine and tyrosine specific phosphatases. ser/thr phosphatases are divided into four subclasses (pp1, pp2a, pp2b, pp2c) on the basis of their substrate specificity, metal ion dependence and inhibitor sensitivity. we were able to detect the ac ...19979406609
mating types and sexual development in filamentous ascomycetes.the progress made in the molecular characterization of the mating types in several filamentous ascomycetes has allowed us to better understand their role in sexual development and has brought to light interesting biological problems. the mating types of neurospora crassa, podospora anserina, and cochliobolus heterostrophus consist of unrelated and unique sequences containing one or several genes with multiple functions, related to sexuality or not, such as vegetative incompatibility in n. crassa ...19979409146
identification and characterization of a novel arabinoxylanase from wheat endogenous wheat (triticum aestivum) flour endoxylanase was purified to homogeneity from a crude wheat flour extract by ammonium sulfate precipitation and cation-exchange chromatography. the 30-kd protein had an isoelectric point of 9.3 or higher. a sequence of 19 amino acids at the nh2 terminus showed 84.2% identity with an internal sequence of 15-kd grain-softness protein, friabilin. high-performance anion-exchange chromatography and gel-permeation analysis of the hydrolysis products indica ...19979414565
cloning and characterization of two phosphate transporters from medicago truncatula roots: regulation in response to phosphate and to colonization by arbuscular mycorrhizal (am) fungi.most vascular plants can acquire phosphate from the environment either directly, via the roots, or indirectly, via a fungal symbiont that invades the cortical cells of the root. here we have identified two cdna clones (mtpt1 and mtpt2) encoding phosphate transporters from a mycorrhizal root cdna library (medicago truncatula/glomus versiforme). the cdnas represent m. truncatula genes and the encoded proteins share identity with high-affinity phosphate transporters from arabidopsis, potato, yeast, ...19989425684
mating type in filamentous fungi.mating type genes regulate sexual compatibility and sexual reproduction in fungi. this review focuses on recent molecular analyses of well-characterized mating systems from representative ascomycete (neurospora crassa, podospora anserina) and basidiomycete (ustilago maydis, coprinus cinereus, schizophyllum commune) fungi. these mating systems include many conserved components, such as gene regulatory polypeptides and pheromone/receptor signal transduction cascades, as well as conserved processes ...19979442896
identification of the tyky homologous subunit of complex i from neurospora crassa.a polypeptide subunit of complex i from neurospora crassa, homologous to bovine tyky, was expressed in escherichia coli, purified and used for the production of rabbit antiserum. the mature mitochondrial protein displays a molecular mass of 21280 da and results from cleavage of a presequence consisting of the first 34 n-terminal amino acids of the precursor. this protein was found closely associated with the peripheral arm of complex i.19979452770
umchs5, a gene coding for a class iv chitin synthase in ustilago maydis.a fragment corresponding to a conserved region of a fifth gene coding for chitin synthase in the plant pathogenic fungus ustilago maydis was amplified by means of the polymerase chain reaction (pcr). the amplified fragment was utilized as a probe for the identification of the whole gene in a genomic library of the fungus. the predicted gene product of umchs5 has highest similarity with class iv chitin synthases encoded by the chs3 genes from saccharomyces cerevisiae and candida albicans, chs-4 f ...19979454647
synthesis and differential turnover of the cys3 regulatory protein of neurospora crassa are subject to sulfur control.the transcription factor cys3 of neurospora crassa is a positive regulator of the sulfur regulatory circuit which contains many structural genes involved in sulfur metabolism. expression and degradation of the cys3 protein are precisely regulated in a sulfur-dependent manner. cys-3 expression was found to be fully repressed by high concentrations of methionine or inorganic sulfate present in the culture medium and to be derepressed when these favored sulfur sources were limited. cys-3 transcript ...19989457847
recombinant pseudorabies virus dnase exhibits a recbcd-like catalytic function.the pseudorabies virus (prv) dnase gene has previously been mapped within the prv genome. to characterize further the enzymic properties of prv dnase, this enzyme was expressed in escherichia coli with the use of a pet expression vector. the protein was purified to homogeneity and assayed for nuclease activity in vitro. recombinant prv dnase exhibited an alkaline ph preference and an absolute requirement for mg2+ ions that could not be replaced by ca2+ and na+ ions. further studies showed that p ...19989461490
differential expression of glucose-regulated (grp78) and heat-shock-inducible (hsp70) genes during asexual development of neurospora crassa.the expression of a glucose-regulated gene (grp78) changes significantly during the vegetative life cycle of neurospora crassa: the amounts of grp78 mrna are low in dormant conidia, increase during germination and exponential growth, decline in young aerial hyphae and reach a maximum in late (15-18 h) aerial hyphae. heat shock (30 min at 45 degrees c) elevated the mrna level of this gene especially in early aerial hyphae, whereas no increase above the high constitutive amount was found after hea ...19989467899
the neurospora aab-1 gene encodes a ccaat binding protein homologous to yeast hap5.the expression of the am (glutamate dehydrogenase) gene is dependent upon two upstream activating sequences, designated ursam(alpha) and ursam(beta). a heteromeric nuclear protein am alpha binding protein (aab) binds specifically to a ccaat box within the ursam(alpha) element. aab appears to be composed of three components. we used polyclonal antiserum raised against the highly purified aab1 subunit to isolate a partial aab-1 cdna clone, which was then used to isolate a full-length cdna and a ge ...19989475726
hypothesis--origin of the parietal cell: microorganism?parietal cells, which exist in the stomach and in meckel's diverticulum of the ileum, secrete hydrochloric acid by means of h+,k(+)-atpase (alpha- and beta-subunits) in the human body. although parietal cells are generated from progenitor cells (stem cells), their lifespan (approximately 150-200 days) is distinctly longer than that of surface epithelial cells (4 days), which are also derived from stem cells. microorganisms, including bacteria (both aerobic and anaerobic), are able to pump h+ out ...19979479641
an additional role for the f-box motif: gene regulation within the neurospora crassa sulfur control network.the f-box represents a protein motif originally identified as a conserved amino-terminal domain within the neurospora crassa negative regulator sulfur controller-2. recently, f-boxes have been found within a number of cell cycle regulatory proteins, where they mediate ubiquitin-driven proteolytic events required for major cell cycle transitions. f-box function, however, is not restricted solely to cell cycle pathways. here we present evidence expanding f-box function to encompass gene regulatory ...19989482900
carbon allocation in ectomycorrhizas: identification and expression analysis of an amanita muscaria monosaccharide transporter.ectomycorrhizas are formed between certain soil fungi and fine roots of predominantly woody plants. an important feature of this symbiosis is the supply of plant-derived carbohydrates to the fungus. as a first step toward a better understanding of the molecular basis of this process, we cloned a monosaccharide transporter from the ectomycorrhizal fungus amanita muscaria. degenerate oligonucleotide primers were designed to match conserved regions from known fungal sugar transporters. a cdna fragm ...19989487692
a nima homologue promotes chromatin condensation in fission yeast.entry into mitosis requires p34(cdc2), which activates downstream mitotic events through phosphorylation of key target proteins. in aspergillus nidulans, the nima protein kinase has been identified as a potential downstream target and plays a role in regulating chromatin condensation at mitosis. nima- mutants arrest in a state that physically resembles interphase even though p34(cdc2) is fully active. despite evidence for the existence of nima-like activities in a variety of cell types, the only ...19989490640
isolation of casln1 and canik1, the genes for osmosensing histidine kinase homologues, from the pathogenic fungus candida albicans.recent studies have revealed that fungi possess a mechanism similar to bacterial two-component systems to respond to extracellular changes in osmolarity. in saccharomyces cerevisiae, sln1p contains both histidine kinase and receiver (response regulator) domains and acts as an osmosensor protein that regulates the downstream hog1 map kinase cascade. sln1 of candida albicans was functionally cloned using an s. cerevisiae strain in which sln1 expression was conditionally suppressed. deletion analys ...19989493379
cloning and characterization of a general amino acid control transcriptional activator from the chestnut blight fungus cryphonectria parasitica.we have cloned and characterized a homologue of the neurospora crassa general amino acid control gene cpc-1 from the chestnut blight fungus cryphonectria parasitica. the deduced amino acid sequence of c. parasitica cpc1 (cpcpc1) contains regions with significant homology to the transcriptional activation, dna binding, and dimerization domains previously defined for n. crassa cpc1 (nccpc1) and the equivalent "b-zip" transcription factor from saccharomyces cerevisiae, gcn4 (scgcn4). treatment of c ...19989501479
consistent structure between bacterial and mitochondrial nadh:ubiquinone oxidoreductase (complex i).respiratory chains of bacteria and mitochondria contain closely related forms of the proton-pumping nadh:ubiquinone oxidoreductase (complex i). in bacteria the complex has a molecular mass of approximately 530 kda and consists of 14 different subunits. the homologues of these 14 subunits together with some 27 additional subunits make up the mitochondrial complex, adding up to a molecular mass of approximately 1 mda. we calculated three-dimensional models at medium resolution of isolated and nega ...19989514725
the aspergillus nidulans sulphur regulatory gene sconb encodes a protein with wd40 repeats and an f-box.the aspergillus nidulans gene sconb, one of the four identified genes controlling sulphur metabolite repression, was cloned and analysed. it encodes a polypeptide of 678 amino acids containing seven wd repeats characteristic of the large wd40 family of eukaryotic regulatory proteins. the sconb protein has nuclear localisation signals and is very similar to the neurospora crassa scon2 and saccharomyces cerevisiae met30 proteins, both of which are involved in the regulation of sulphur metabolism. ...19989520259
the role of the dynactin complex in intracellular motility.dynactin is a multisubunit complex that binds to the minus-end-directed microtubule motor cytoplasmic dynein and may provide a link between the motor and its cargo. results from genetic studies in saccharomyces cerevisiae, neurospora crassa, aspergillus nidulans, and drosophila have suggested that cytoplasmic dynein and dynactin function in the same cellular pathways. p150glued, a vertebrate homologue of the drosophila gene glued, is the largest polypeptide in the dynactin complex with multiple ...19989522459
secondary structure of the intact h+,k+-atpase and of its membrane-embedded region. an attenuated total reflection infrared spectroscopy, circular dichroism and raman spectroscopy study.models of p-type atpase predict that membrane-embedded fragments represent about 20% of the protein and adopt an all-alpha-helical structure. while this prediction was confirmed for the ca2-atpase [corbalan-garcia, s., teruel, j., villalain, j. & gomez-fernandez, j. (1994) biochemistry 33, 8247-8254], it is at odds with recent experimental evidence gathered on the neurospora crassa plasma membrane h+-atpase [vigneron, l., ruysschaert, j.-m. & goormaghtigh, e. (1995) j. biol. chem. 270, 17685-176 ...19989523697
molecular genetics of mating recognition in basidiomycete fungi.the recognition of compatible mating partners in the basidiomycete fungi requires the coordinated activities of two gene complexes defined as the mating-type genes. one complex encodes members of the homeobox family of transcription factors, which heterodimerize on mating to generate an active transcription regulator. the other complex encodes peptide pheromones and 7-transmembrane receptors that permit intercellular signalling. remarkably, a single species may have many thousands of cross-compa ...19989529887
characterization of the aspergillus nidulans nmra gene involved in nitrogen metabolite repression.the gene nmra of aspergillus nidulans has been isolated and found to be a homolog of the neurospora crassa gene nmr-1, involved in nitrogen metabolite repression. deletion of nmra results in partial derepression of activities subject to nitrogen repression similar to phenotypes observed for certain mutations in the positively acting area gene.19989537404
the neurospora rca-1 gene complements an aspergillus flbd sporulation mutant but has no identifiable role in neurospora sporulation.the aspergillus nidulans flbd gene encodes a protein with a myb-like dna-binding domain that is proposed to act in concert with other developmental regulators to control initiation of conidiophore development. we have identified a neurospora crassa gene called rca-1 (regulator of conidiation in aspergillus) based on its sequence similarity to flbd. we found that n. crassa rca-1 can complement the conidiation defect of an a. nidulans flbd mutant and that induced expression of rca-1 caused conidia ...19989539422
molecular analysis of a glucose-regulated gene (grp78) of neurospora crassa.the nucleotide sequence of the gene encoding the glucose-regulated protein 78 (grp78) of neurospora crassa was determined. the orf codes for a protein of 662 amino acids (72 kda) and belongs to the heat shock protein 70 (hsp70) gene family, which is characterized by three hsp70 'signature sequences'. the grp78 gene contains 5 introns. the protein carries the er retention signal hdel at its carboxy terminus and is most homologous to the kar2/grp78 protein of saccharomyces cerevisiae (78%) and to ...19989545520
oxidation of catalase by singlet oxygen.different bands of catalase activity in zymograms (cat-1a-cat-1e) appear during neurospora crassa development and under stress conditions. here we demonstrate that singlet oxygen modifies cat-1a, giving rise to a sequential shift in electrophoretic mobility, similar to the one observed in vivo. purified cat-1a was modified with singlet oxygen generated from a photosensitization reaction; even when the reaction was separated from the enzyme by an air barrier, a condition in which only singlet oxy ...19989553125
the intriguing evolution of the "b" and "g" subunits in f-type and v-type atpases: isolation of the vma-10 gene from neurospora crassa.we have characterized the vma-10 gene which encodes the g subunit of the vacuolar atpase in neurospora crassa. the gene is somewhat unusual in filamentous fungi because it contains five introns, comprising 71% of the region between the translation start and stop codons. the 5' untranslated region of the gene contains several elements that have been identified in other genes that encode subunits of the vacuolar atpase in n. crassa. a comparison of g subunits from n. crassa, s. cerevisiae, and ani ...19979559854
the fluffy gene of neurospora crassa encodes a gal4p-type c6 zinc cluster protein required for conidial development.neurospora crassa fluffy (fl) mutants are unable to produce macroconidia. we cloned the fl gene to determine its role in regulating conidiation. a cosmid clone containing fl was identified by complementation. the sequence of fl revealed that it encodes a gal4p-type c6 zinc cluster protein with greatest similarity to the n. crassa nit4 protein that regulates genes required for nitrate utilization. analysis of several fl mutant alleles demonstrated that null mutants are blocked in the budding phas ...19989560395
cloning and characterization of the yeast rad1 homolog gene (mus-38) from neurospora crassa: evidence for involvement in nucleotide excision repair.a neurospora crassa gene encoding a product with homology to the saccharomyces cerevisiae rad1 nucleotide excision repair (ner) protein was isolated by degenerate pcr. the predicted protein consists of 892 amino acids with a molecular weight of 100.4 kda, and 32-37% identity to the xpf/ercc4 protein family. the homolog was mapped to the left arm of linkage group i, the location of the mus-38 gene. subsequently, gene inactivation and complementation studies identified the rad1 homolog as mus-38. ...19989560435
three-dimensional structure of bovine nadh:ubiquinone oxidoreductase (complex i) at 22 a in ice.nadh:ubiquinone oxidoreductase (complex i) is the first and largest complex in the electron transport chain of mitochondria. the bovine complex purified from cardiac muscle consists of at least 42 different subunits with a combined molecular mass of about 890 kda. the three-dimensional structure of the complex was determined at 22 a from single particles embedded in vitrified ice using electron cryo-microscopy. the structure was calculated using a new program to align particles, to correct for t ...19989571020
repeat-induced point mutations in pad-1, a putative rna splicing factor from neurospora crassa, confer dominant lethal effects on ascus development.we describe the characterization of a gene, pad-1, from neurospora crassa which displays sequence characteristics of the rs class of hnrna-binding proteins (hnrnp) and mrna splicing factors. this is the first report of the isolation of a putative hnrnp gene from n. crassa. pad-1 showed 30% identity and 57% similarity to a protein, hcc1, which was isolated using autoantibodies from patients suffering from hepatocellular carcinoma. both hcc1 and pad-1 show amino acid sequence similarities to the h ...19989578630
role of fungal dynein in hyphal growth, microtubule organization, spindle pole body motility and nuclear migration.cytoplasmic dynein is a microtubule-associated motor protein with several putative subcellular functions. sequencing of the gene (dhc1) for cytoplasmic dynein heavy chain of the filamentous ascomycete, nectria haematococca, revealed a 4,349-codon open reading frame (interrupted by two introns) with four highly conserved p-loop motifs, typical of cytoplasmic dynein heavy chains. the predicted amino acid sequence is 78.0% identical to the cytoplasmic dynein heavy chain of neurospora crassa, 70.2% ...19989580563
the isolation and characterization of nrc-1 and nrc-2, two genes encoding protein kinases that control growth and development in neurospora crassa.using an insertional mutagenesis approach, a series of neurospora crassa mutants affected in the ability to control entry into the conidiation developmental program were isolated. one such mutant, gth16-t4, was found to lack normal vegetative hyphae and to undergo constitutive conidiation. the affected gene has been named nrc-1, for nonrepressible conidiation gene #1. the nrc-1 gene was cloned from the mutant genomic dna by plasmid rescue, and was found to encode a protein closely related to the ...19989584090
role of the negative charges in the cytosolic domain of tom22 in the import of precursor proteins into mitochondria.tom22 is an essential mitochondrial outer membrane protein required for the import of precursor proteins into the organelles. the amino-terminal 84 amino acids of tom22 extend into the cytosol and include 19 negatively and 6 positively charged residues. this region of the protein is thought to interact with positively charged presequences on mitochondrial preproteins, presumably via electrostatic interactions. we constructed a series of mutant derivatives of tom22 in which 2 to 15 of the negativ ...19989584158
cell wall active antifungal compounds produced by the marine fungus hypoxylon oceanicum ll-15g256. iii. biological properties of 15g256 gamma.15g256 gamma is a cyclic lipopeptide antifungal agent discovered in a mechanism of action screen for cell wall acting antifungal agents. the compound shows moderate activity in both greenhouse tests against plant disease caused by pathogenic fungi and in in vitro tests against human fungal pathogens. microscopic examination of treated fungi suggests that the compound acts by the inhibition of cell wall biosynthesis. however, in vitro inhibition of neurospora crassa glucan and chitin synthase wer ...19989589067
bacterial expression and characterization of the mitochondrial outer membrane channel. effects of n-terminal modifications.several forms of the voltage-dependent anion-selective channel (vdac) have been expressed at high yield in escherichia coli. full-length constructs of the proteins of neurospora crassa and saccharomyces cerevisiae (ncvdac and scvdac) have been made with 20-residue-long, thrombin-cleavable, his6-containing n-terminal extensions. ncvdac purified from bacteria or mitochondria displays a far-uv cd spectrum (in 1% lauryl dimethylamine oxide at ph 6-8) similar to that of bacterial porins, indicating e ...19989593723
complex i from the fungus neurospora crassa.respiratory chain complex i is a complicated enzyme of mitochondria, that couples electron transfer from nadh to ubiquinone to the proton translocation across the inner membrane of the organelle. the fungus neurospora crassa has been used as one of the main model organisms to study this enzyme. complex i is composed of multiple polypeptide subunits of dual genetic origin and contains several prosthetic groups involved in its activity. most subunits have been cloned and those binding redox centre ...19989593837
transcriptional activation of the aspergillus nidulans gpda promoter by osmotic signals.a differentially expressed gpda cdna clone was isolated from nacl-adapted aspergillus nidulans (fgsc359) and identified as glyceraldehyde-3-phosphate dehydrogenase (gpda) on the basis of its nucleotide sequence. the level of gpda rna substantially increased in cultures gradually adapted to nacl but was greatly reduced in cultures exposed briefly to a high concentration of nacl. a pyrg auxotroph of a. nidulans (a773) was cotransformed with a gpda-uida construct and a plasmid containing the neuros ...19989603839
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