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isolation and characterization of a laccase gene from podospora anserina.the genome of the filamentous ascomycete podospora anserina contains at least four non-adjacent regions that are homologous to the laccase gene of neurospora crassa. one of these regions contains a gene (lac2) encoding a protein that displays 62% identity with the n. crassa laccase. in shaken cultures, lac2 mrna is present at low basal levels throughout the growth phase but increases at least 20-fold at the beginning of the autolytic phase and decreases again thereafter. addition of aromatic xen ...19968914515
nuc-2, a component of the phosphate-regulated signal transduction pathway in neurospora crassa, is an ankyrin repeat protein.in response to phosphorus limitation, the fungus neurospora crassa synthesizes a number of enzymes that function to bring more phosphate into the cell. the nuc-2 protein appears to sense the availability of phosphate and transmits the signal downstream to the regulatory pathway. the nuc-2+ gene has been cloned by its ability to restore growth of a nuc-2 mutant under restrictive conditions of high ph and low phosphate concentration. we mapped the cloned gene to the right arm of linkage group ii, ...19968917314
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 4.1.3.21). 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
site-directed mutagenesis of the cysteine residues in the neurospora crassa plasma membrane h(+)-atpase.a high-yield yeast expression system for site-directed mutagenesis of the neurospora crassa plasma membrane h(+)-atpase has recently been reported (mahanty, s. k., rao, u. s., nicholas, r. a., and scarborough, g. a. (1994) j. biol. chem. 269, 17705-17712). using this system, each of the eight cysteine residues in the atpase was changed to a serine or an alanine residue, producing strains c148s and c148a, c376s and c376a, c409s and c409a, c472s and c472a, c532s and c532a, c545s and c545a, c840s a ...19968550588
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 fungi.in 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 effects.an 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 crassa.in 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
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
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
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 fungi.contact-sensing 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 sources.in 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 flour.an 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
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
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
carbon-13 nuclear-magnetic-resonance spectroscopy of whole cells and of cytochrome c from neurospora crass grown with (s-me-13c)methionine.neurospora crassa cytochrome c biosynthetically labelled with [s-me-13c]methionine was prepared and analysed by 13c nuclear-magnetic-resonance spectroscopy. the methyl group of methionine is extensively incorporated into an n-trimethyl-lysine-72 residue arise from s-adenosylmethionine transmethylation, and that the methyl carbons of methionine residues are sufficiently close to the haem centre to experience chemical shifts from the ring currents of the tetrapyrrole pi electrons and broadening du ...1997179524
oxidation processes and ubiquinone localization in the branched respiratory system of mi-1 mutant of neurospora crassa.1. stimulation of succinate oxidation in mi-1 mitochondria by mg2+ and pi is abolished on uncoupling, which points to the energy-linked activation of succinate oxidation. 2. mitochondria exhibited respiratory control with succinate and nadh when the cyanide-insensitive oxidation was inhibited by salicylhydroxamic acid (sham). sham lowered the oxidation rate with nadh and succinate to the same level, though the nadh oxidation rate was 2.5 times as high as with succinate. 3. despite the high stimu ...1997168706
studies on the poky mutant of eurospora crassa. fingerprint analysis of mitochondrial ribosomal rna.base sequence and methylation of mitochondrial ribosomal rnas from wild type and poky strains of neurospora crassa were compared to determine whether a mutational lesion exists in poky 19 s rna. at the outset, new procedures were developed for the isolation of intact nucleic acids from neurospora mitochondria based on the substitution of ca2+ for mg2+ in the isolation media to inhibit mitochondrial nuclease activity. using these procedures, intact and highly purified 32p-labeled ribosomal rnas w ...1997178661
homology modeling of neurospora crassa geranylgeranyl pyrophosphate synthase: structural interpretation of mutant phenotypes.a model of the tertiary structure of the neurospora crassa carotenogenic prenyltransferase, geranylgeranyl pyrophosphate synthase (ggpps), is presented, based on structural homology with other prenyltransferases and on the crystal structure of recombinant avian farnesyl pyrophosphate synthase (fpps). the conserved aspartate-rich motifs ddxx(xx)d and associated basic residues, considered to be the active sites for binding and catalysis in all prenyltransferases, are highly conserved in the n. cra ...19979464568
developmental and photoregulation of three neurospora crassa carotenogenic genes during conidiation induced by desiccation.neurospora crassa asexual sporulation (conidiation) is induced by different cues including desiccating aerial environments. three of the genes that are expressed during this developmental pathway are the albino (al) genes, which encode carotenoid biosynthetic enzymes. if conidiation is induced by nutrient deprivation in liquid culture, two overlapping al-3 mrnas, al-3 (m) and al-3 (c), are expressed (arpaia et al., 1995, dev. biol. 170, 620-635). here we quantitate accumulation of each of the fo ...19979126619
characterization of vacuolar arginine uptake and amino acid efflux in neurospora crassa using cupric ion to permeabilize the plasma membrane.treatment of neurospora crassa mycelia with cupric ion has been shown to permeabilize the plasma and mitochondrial membranes. permeabilized mycelia were shown to take up arginine into the vacuoles. uptake was atp-independent and appeared to be driven by an existing k+-gradient. the kinetic characteristics of the observed uptake were similar to those observed using vacuolar membrane vesicles: the km for arginine uptake was found to be 4.2-4.5 mm. permeabilized mycelia were used to study the regul ...19979454645
blue light regulation in neurospora crassa.the fungus neurospora crassa has been shown to be a paradigm for photobiological, biochemical, and genetic studies of blue light perception and signal transduction. several different developmental and morphological processes of neurospora are regulated by blue light and can be divided into early and late blue light responses. the characterization of two central regulator proteins of blue light signal transduction in neurospora crassa, wc1 and wc2, and the isolation of light-regulated genes, indi ...19979454641
effects of the tol mutation on allelic interactions at het loci in neurospora crassa.a mutant at the tol locus of neurospora crassa can suppress heterokaryon (vegetative) incompatibility associated with differences at the mating-type locus. we tested the ability of this allele to suppress the vegetative incompatibility reactions that can occur when strains differ at one of nine het loci (het-c, -d, -e, -5, -6, -7, -8, -9, and -10). we found no cases in which the tol mutant suppresses a heteroallelic het locus interaction. thus, the interaction(s) that leads to vegetative incompa ...19979449795
specific, high affinity binding sites for an antifungal plant defensin on neurospora crassa hyphae and microsomal membranes.hs-afp1, an antifungal plant defensin from seed of the plant heuchera sanguinea, was radioactively labeled using t-butoxycarbonyl-[35s]l-methionine n-hydroxysuccinimidyl ester, resulting in a 35s-labeled peptide with unaltered antifungal activity. [35s]hs-afp1 was used to assess binding on living hyphae of the fungus neurospora crassa. binding of [35s]hs-afp1 was found to be competitive, reversible, and saturable with an apparent kd of 29 nm and a bmax of 1.4 pmol/mg protein. [35s]hs-afp1 also b ...19979405418
genetic analysis of signal transduction through light-induced protein phosphorylation in neurospora crassa perithecia.we have previously demonstrated that blue light induces the phosphorylation of a 15-kda protein in crude membrane fractions of neurospora crassa mycelia. here we report the isolation and characterization of a mutant (psp; phosphorylation of small proteins that is completely defective for phosphorylation of that protein, as assayed in both crude membrane and soluble fractions. this mutation defines a unique locus that maps to linkage group vr between al-3 and his-6. to elucidate the photobiologic ...19979435784
two distinct protein-protein interactions between the nit2 and nmr regulatory proteins are required to establish nitrogen metabolite repression in neurospora crassa.nitrogen metabolism is a highly regulated process in neurospora crassa. the structural genes that encode nitrogen catabolic enzymes are subject to nitrogen metabolite repression, mediated by the positive-acting nit2 protein and by the negative-acting nmr protein. nit2, a globally acting factor, is a member of the gata family of regulatory proteins and has a single cys2/cys2 zinc finger dna-binding domain. the negative-acting nmr protein interacts via specific protein-protein binding with two dis ...19979427402
white collar proteins: passing the light signal in neurospora crassa.the filamentous fungus neurospora crassa is an excellent paradigm for the study of blue light signal transduction. the isolation and characterization of the genes for two central regulators of the blue light response, white collar-1 and white collar-2, have begun to shed light on the mechanism of blue light signal transduction in fungi. these proteins are not only proposed to encode blue-light-activated transcription factors but also to be elements of the blue light signal transduction pathway.19979402704
differential hsc70 expression during asexual development of neurospora crassa.the constitutive and the heat-shock-induced expression of members of heat-shock protein families changed during vegetative development and conidiation of neurospora crassa as determined by two-dimensional gel electrophoresis. western blot and elisa analyses revealed the highest amounts of the constitutive heat-shock protein 70 (hsc70) in conidiating aerial hyphae and dormant conidia. during conidial germination the amount of hsc70 decreased and subsequently increased during vegetative growth. st ...19979387240
site and significance of cysteine residues in xylose reductase from neurospora crassa as deduced by fluorescence studies.inactivation of xylose reductase (xr) by p-hydroxy-mercury benzoate (phmb) was found to be biphasic with second-order rate constants of 80 and 6 m-1s-1 for the fast (kf) and slow (ks) phase respectively. spectroscopic studies indicated that the inactivation was due to modification of one cys residue per molecule of xr and not due to subsequent disruption of the quaternary structure. the binding of nadph to xr (kd 0.9 microm) was depressed on modification of the enzyme by phmb (kd 2.3 microm). th ...19979367847
primary structure of neurospora crassa gamma-tubulin.we report the isolation and characterization of a cdna-clone for gamma-tubulin from n. crassa. the deduced amino-acid sequence of the cdna-clone most resembles that of gamma-tubulins from other fungi. differential southern blot analysis demonstrates that most probably a single gene represents gamma-tubulin. in addition, northern blot analysis shows that most probably only one single mrna encoding gamma-tubulin is transcribed in n. crassa. in a phylogenetic tree that includes all known gamma-tubu ...19979358070
high-affinity no(3-)-h+ cotransport in the fungus neurospora: induction and control by ph and membrane voltage.high-affinity nitrate transport was examined in intact hyphae of neurospora crassa using electrophysiological recordings to characterize the response of the plasma membrane to no3- challenge and to quantify transport activity. the no3(-)-associated membrane current was determined using a three electrode voltage clamp to bring membrane voltage under experimental control and to compensate for current dissipation along the longitudinal cell axis. nitrate transport was evident in hyphae transferred ...19979351892
evidence for a cysteine-histidine thioether bridge in functional units of molluscan haemocyanins and location of the disulfide bridges in functional units d and g of the betac-haemocyanin of helix pomatia.in functional units d and g from the betac-haemocyanin of the gastropod helix pomatia, aminoacid analysis in the presence of dimethyl sulfoxide showed the occurrence of seven cysteine residues. titration with 5,5'-dithiobis(2-nitrobenzoic acid), however, did not reveal any free thiol group. pepsinolysis at ph 2.0 followed by amino acid analysis and partial sequencing of the cysteine-containing peptides showed that six cysteine residues are involved in the formation of three disulfide bridges at ...19979342242
dna methylation inhibits elongation but not initiation of transcription in neurospora crassa.in plants, animals, and fungi, dna methylation is frequently associated with gene silencing, yet little is known about the role of the methylation in silencing. in neurospora crassa, repeated sequences are silenced by repeat-induced point mutation (rip) and genes that have suffered numerous gc --> at mutations by rip are typically methylated at remaining cytosines. we investigated possible effects on transcription from methylation associated with rip by taking advantage of 5-azacytidine, which p ...19979308966
temperature compensation of the circadian period length--a special case among general homeostatic mechanisms of gene expression?in neurospora crassa, as well as in other organisms, the expression of housekeeping genes is transiently suppressed after exposure to higher temperatures (30-45 degrees c); expression is then reactivated and adapts after a few-hours to values closer to the initial rates. adaptive mechanisms apparently exist in the processes of transcription, rna processing, and translation and render protein synthesis rates temperature compensated. heat shock proteins (hsps) play an important role within these m ...19979298284
temperature compensation and membrane composition in neurospora crassa.the link between temperature compensation of the circadian rhythm and temperature-induced adjustment of membrane composition in neurospora crassa is briefly reviewed. in common with most organisms, neurospora responds to changes in growth temperature by adjusting its lipid composition, primarily by increasing the degree of unsaturation of its fatty acids at low temperature. this may result in maintenance of either membrane fluidity or phase transition behavior over a range of temperatures. in ne ...19979298281
isolation of quelling-defective (qde) mutants impaired in posttranscriptional transgene-induced gene silencing in neurospora crassa.we report the isolation of 15 neurospora crassa mutants defective in "quelling" or transgene-induced gene silencing. these quelling-defective mutants (qde) belonging to three complementation groups have provided insights into the mechanism of posttranscriptional gene silencing in n. crassa. the recessive nature of the qde mutations indicates that the encoded gene products act in trans. we show that when qde genes are mutated in a transgenic-induced silenced strain containing many copies of the t ...19979294193
overlapping functions for two g protein alpha subunits in neurospora crassa.heterotrimeric g proteins, consisting of alpha, beta and gamma subunits, mediate a variety of signaling pathways in eukaryotes. we have previously identified two genes, gna-1 and gna-2, that encode g protein alpha subunits in the filamentous fungus neurospora crassa. mutation of gna-1 results in female infertility and sensitivity to hyperosmotic media. in this study, we investigate the expression and functions of gna-2. results from western analysis and measurements of gna-2 promoter-lacz fusion ...19979286674
ribosome stalling is responsible for arginine-specific translational attenuation in neurospora crassa.the neurospora crassa arg-2 upstream open reading frame (uorf) plays a role in negative arginine-specific translational regulation. primer extension inhibition analyses of arg-2 uorf-containing rna translated in a cell-free system in which arginine-specific regulation was retained revealed "toeprints" corresponding to ribosomes positioned at the uorf initiation and termination codons and at the downstream initiation codon. at high arginine concentrations, the toeprint signal corresponding to rib ...19979271370
heat shock protein 80 of neurospora crassa, a cytosolic molecular chaperone of the eukaryotic stress 90 family, interacts directly with heat shock protein 70.the subunit structure of hsp80, the most abundant heat-shock protein of neurospora crassa, was examined by chemical cross-linking of the purified protein in vitro. resolution of glutaraldehyde-treated hsp80 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis sds-page suggests that the native state of this protein is a tetramer; the relative proportion of cross-linked species, estimated by the fraction of protein recovered in each category, is consistent with a dimer-of-dimer structure. ...19979254620
allelic specificity at the het-c heterokaryon incompatibility locus of neurospora crassa is determined by a highly variable domain.in filamentous fungi, the ability to form a productive heterokaryon with a genetically dissimilar individual is controlled by specific loci termed het loci. only strains homozygous for all het loci can establish a heterokaryon. in neurospora crassa, 11 loci, including the mating-type locus, regulate the capacity to form heterokaryons. an allele of the het-c locus (het-cor) of n. crassa has been previously characterized and encodes a nonessential 966 amino acid glycine-rich protein. herein, we de ...19979258675
ppt-1, a neurospora crassa ppt/pp5 subfamily serine/threonine protein phosphatase.we isolated a n. crassa cdna clone encoding a novel-type serine/threonine phosphatase. the gene (mapped to lgvr), designated ppt-1, encodes a 479 amino acid putative polypeptide which contains a conserved tetratricopeptide repeat (tpr) motif. ppt-1 transcript levels are abundant in conidia and decrease during germination, indicating that ppt-1 is developmentally regulated.19979256060
novel pattern of dna methylation in neurospora crassa transgenic for the foreign gene hph.it has previously been reported that multiple copies of the hph gene integrated into the genome of neurospora crassa are methylated at hpa ii sites (ccgg) during the vegetative life cycle of the fungus, while hph genes integrated as single copies are not methylated. furthermore, methylation is correlated with silencing of the gene. we report here the methylation state of cytosine residues of the major part of the promoter region of the hph gene integrated into the genome of the multiple copy str ...19979171093
mutations of pma-1, the gene encoding the plasma membrane h+-atpase of neurospora crassa, suppress inhibition of growth by concanamycin a, a specific inhibitor of vacuolar atpases.concanamycin a (cca), a specific inhibitor of vacuolar atpases, inhibited growth of neurospora crassa in medium adjusted to ph 7 or above. mutant strains were selected for growth on medium containing 1.0 microm cca. sixty-four (of 66) mutations mapped in the region of the pma1 locus, which encodes the plasma membrane h+-atpase. analysis of v-atpase activity in isolated vacuolar membranes from the mutant strains showed wild-type activity and sensitivity to cca. in contrast, plasma membrane h+-atp ...19979169444
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