TitleAbstractYear(sorted descending)
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
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
pronounced cytoplasmic ph gradients are not required for tip growth in plant and fungal cells.the existence of pronounced cytoplasmic ph gradients within the apices of tip-growing cells, and the role of cytoplasmic ph in regulating tip growth, were investigated in three different cell types: vegetative hyphae of neurospora crassa; pollen tubes of agapanthus umbellatus; and rhizoids of dryopteris affinis gametophytes. examination of cytoplasmic ph in growing cells was performed by simultaneous, dual emission confocal ratio imaging of the ph-sensitive probe carboxy snarf-1. considerable at ...19979191043
a putative role for calmodulin in the activation of neurospora crassa chitin synthase.the possible role of calmodulin in cell wall formation and chitin synthesis was studied in neurospora crassa by examining the effects of anti-calmodulin agents on protoplast regeneration and possible associations between chitin synthase and calmodulin related proteins in microsomal isolates. protoplast regeneration was inhibited by trifluoperazine (> 20 microm), an anticalmodulin agent. chitin synthase activity in microsomes was associated with that of calmodulin-dependent protein kinase and inh ...19979163912
mutation of the cys-9 gene, which encodes thioredoxin reductase, affects the circadian conidiation rhythm in neurospora crassa.ten cysteine auxotrophs of neurospora crassa were examined with regard to the period lengths of their circadian conidiation rhythms. one of the these cysteine auxotrophs, cys-9, showed dramatic changes in the circadian conidiation rhythm. at 10 microm methionine, the cys-9 mutant had a period length that was 5 hr shorter than that of the wild-type strain during the first 3 days after transfer to continuous darkness. at this concentration of methionine, the period length was unstable after the fo ...19979136004
inhibition of neurospora crassa growth by a glucan synthase-1 antisense construct.we have used the filamentous fungus, neurospora crassa, as a model system to test the concept that antisense targeting of the cell-wall assembly enzyme, (1,3)beta-glucan synthase [e.c.; udpglucose: 1,3-beta-d-glucan 3-beta-d-glucosyltransferase], leads to a corresponding decrease in growth of the organism. previously, our laboratory isolated a gene (glucan synthase-1, gs-1) that is required for (1,3)beta-glucan synthase activity. wild-type cells were transformed with dna vectors encodin ...19979099631
a ras homologue of neurospora crassa regulates order to study the role of signal transduction pathways in the regulation of morphology in neurospora crassa, we cloned and characterized a ras homologue, termed nc-ras2. the predicted protein product of this gene is composed of 229 amino acid residues and contains all the consensus sequences shared by the ras protein family. the gene is located in linkage group v. an nc-ras2 disruptant showed morphological characteristics very similar to those of the smco7 mutant, which also maps to linkage ...19979180696
a gene required for nuclear migration in neurospora crassa codes for a protein with cysteine-rich, lim/ring-like domains.the ro mutants of neurospora crassa are defective in nuclear migration and hyphal morphogenesis. several of the ro loci have recently been shown to encode components of the dynein/dynactin motor complex. here we report on the cloning and characterization of the ro-2 gene which codes for a novel 80 kda protein that has two cys-rich motifs which resemble zinc-binding lim or ring domains thought to mediate protein-protein interactions. ro2 also contains several potential binding sites for src homol ...19979159520
identification of genes induced in alkaloid-producing cultures of claviceps order to identify genes which are expressed during alkaloid synthesis in an axenic culture of claviceps sp. (strain atcc 26245), a cdna library from a producing culture was differentially screened with cdna from producing (cdna+) and non-producing (cdna-) cultures, respectively. altogether, ten cdna clones were obtained, the alkaloid-synthesis-correlated expression of which was confirmed by northern analyses. evaluation of their nucleotide and derived amino-acid sequences identified one gene ...19979108144
mutants of neurospora crassa defective in regulation of blue light perception.a new selection system was used to isolate mutants deficient in regulation of blue light perception in neurospora crassa. this selection system has two possible applications and was used for the isolation of either blind or constitutive mutants. we isolated 17 uv-induced mutants that showed the pleiotropic white collar (wc) phenotype and were completely blocked in transduction of the light signal. from the segregation pattern in sexual crosses, we tentatively assigned the 17 mutants to either th ...19979108272
mutagenic activity and specificity of hydrogen peroxide in the ad-3 forward-mutation test in two-component heterokaryons of neurospora the ad-3 forward-mutation test, hydrogen peroxide was at best a weak mutagen in nongrowing conidia from a dna repair-proficient heterokaryon (h-12, uvs-2+/uvs-2+) but was a moderate mutagen in nongrowing conidia from a dna-repair-deficient heterokaryon (h-59, uvs-2/uvs-2) over a narrow range of high concentrations. h-59 also was more sensitive than h-12 to the killing activity of hydrogen peroxide at high concentrations. thus, a dna-repair pathway, of which the gene product of the uvs-2+ alle ...19979100841
purification of a 47-kda calmodulin-binding polypeptide as an actin-binding protein from neurospora crassa.we have enriched a 47-kda polypeptide (p47) from neurospora crassa on the basis of its affinity to calmodulin. the p47 was purified to homogeneity by chromatography on a mono s cation exchange column and evidence is presented that the polypeptide co-sediments specifically with f-actin. the intracellular distribution of p47 and actin was also examined using indirect double immunofluorescence staining of cells at different stages of development. our results suggest that by altering the conformatio ...19979119196
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
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
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
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
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
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
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
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
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
structural analysis of mitochondrial dna molecules from fungi and plants using moving pictures and pulsed-field gel electrophoresis.the size and structure of mitochondrial dna (mtdna) molecules was investigated by conventional and pulsed-field gel electrophoresis (pfge) and by analyzing moving pictures during electrophoresis of individual fluorescently labelled mtdna molecules. little or no mtdna that migrated into the gel was found in circular form for fungi (schizosaccharomyces pombe, saccharomyces cerevisiae and neurospora crassa) or plants (brassica hirta, tobacco, voodoo lily and maize). most mtdna migrated as a smear o ...19968568898
nucleotide sequence and expression of the gene encoding nadp+- dependent glutamate dehydrogenase (gdha) from agaricus bisporus.the gene encoding nadp+-dependent glutamate dehydrogenase (gdha) was isolated from an agaricus bisporus recombinant phage lambda library. the deduced amino acid sequence would specify a 457-amino acid protein that is highly homologous in sequence to those derived from previously isolated and characterized genes coding for microbial nadp+-gdh. the open reading frame is interrupted by six introns. none of the introns is located at either one of the positions of the two introns conserved in the cor ...19968602149
gel mobility shift scanning of the acetate-inducible promoters from neurospora crassa reveals a common co-inducible dna-binding protein.the promoter regions of four acetate-inducible genes of neurospora crassa, acu-3, acu-5, acu-8 and acu-9, have been sequenced. using a scanning gel mobility shift assay particular dna regions in each promoter have been shown specifically to bind partially purified protein extracted from acetate-induced mycelia. the protein-binding regions so defined have common sequence motifs, elements of which are similar to those required for acetate induction in aspergillus nidulans.19968602159
distribution of thiols in microorganisms: mycothiol is a major thiol in most actinomycetes.mycothiol [2-(n-acetylcysteinyl)amido-2-deoxy-alpha-d-glucopyranosyl- (1-->1)-myo-inositol] (msh) has recently been identified as a major thiol in a number of actinomycetes (s. sakuda, z.-y. zhou, and y. yamada, biosci. biotech. biochem. 58:1347-1348, 1994; h. s. c. spies and d. j. steenkamp, eur. j. biochem. 224:203-213, 1994; and g. l. newton, c. a. bewley, t. j. dwyer, r. horn, y. aharonowitz, g. cohen, j. davies, d. j. faulkner, and r. c. fahey, eur. j. biochem. 230:821-825, 1995). since thi ...19968606174
white collar-1, a central regulator of blue light responses in neurospora, is a zinc finger protein.the neurospora crassa blind mutant white collar-1 (wc-1) is pleiotropically defective in all blue light-induced phenomena, establishing a role for the wc-1 gene product in the signal transduction pathway. we report the cloning of the wc-1 gene isolated by chromosome walking and mutant complementation. the elucidation of the wc-1 gene product provides a key piece of the blue light signal transduction puzzle. the wc-1 gene encodes a 125 kda protein whose encoded motifs include a single class four, ...19968612589
characterization of a uv endonuclease gene from the fission yeast schizosaccharomyces pombe and its bacterial homolog.from the fission yeast schizosaccharomyces pombe, a cdna fragment was isolated, which confers uv resistance on repair deficient escherichia coli host cells. the cloned cdna encodes a protein of 68,815 da, which has a 36.6% identity of amino acid sequence with the previously identified 74 kda uv endonuclease of the filamentous fungus neurospora crassa. analysis of several truncated gene constructs shows that only the c-terminal two thirds region, which has 54% identity of amino acid sequence with ...19968614629
molecular cloning and functional expression of human deoxyhypusine synthase cdna based on expressed sequence tag information.deoxyhypusine synthase is an nad(+)-dependent enzyme that catalyses the formation of a deoxyhypusine residue on the eukaryotic initiation factor 5a (eif-5a) precursor by transferring an aminobutyl moiety from spermidine to the epsilon-amino group of a unique lysine residue. we have recently cloned and characterized the neurospora crassa deoxyhypusine synthase cdna using a reverse genetics approach. a genbank search showed that a stretch of the deduced amino acid sequence (96 amino acids) of neur ...19968615810
transient expression of a mitochondrial precursor protein. a new approach to study mitochondrial protein import in cells of higher order to study mitochondrial protein import in the context of whole cell metabolism, we have used the transfection technique based on semliki forest virus (sfv) to express a mitochondrial precursor protein within bhk21 cells and human fibroblasts. recombinant sfv particles mediate a highly efficient, transient transfection of higher eukaryotic cells. the mitochondrial precursor protein used is a fusion protein consisting of the mitochondrial targeting sequence of neurospora crassa atpase subu ...19968620886
an additional copy of the adenylate cyclase-encoding gene relieves developmental defects produced by a mutation in a vegetative incompatibility-controlling gene in podospora identify cellular functions involved in vegetative incompatibility in filamentous fungi, we have initiated the cloning of podospora anserina (pa) mod genes. these genes interfere with the lethal reaction triggered by interaction between incompatible het genes. a gene (pa ac) has been cloned by complementation of developmental defects caused by a mutation in the mod-d gene. this gene encodes a protein of 2145 amino acids (aa)that exhibits strong similarities with many adenylate cyclases (ac). ...19968621071
cloning and characterization of the neurospora crassa cyt-5 gene. a nuclear-coded mitochondrial rna polymerase with a polyglutamine repeat.the neurospora crassa mutants, cyt-5-1 and cyt-5-4 have a cytochrome b- and aa3-deficient phenotype, suggesting that they result from a deficiency in a nuclear-coded component of the mitochondrial gene expression apparatus (bertrand, h., nargang, f. e., colllins, r. a., and zagozeski, c. a. (1977) mol. gen. genet. 153,247-257). the complementing wild-type gene has been cloned and and shown to encode a protein with significant sequence similarity to saccharomyces cerevisiae mitochondrial rna poly ...19968626458
chs-4, a class iv chitin synthase gene from neurospora saccharomyces cerevisiae, most of the cellular chitin is produced by chitin synthase iii, which requires the product encoded by the csd2/cal1/dit101/kt12 gene. we have identified, isolated and structurally characterized as csd2/cal1/dit101/kt12 homologue in the filamentous ascomycete neurospora crassa and have used a "reverse genetics" approach to determine its role in vivo. the yeast gene was used as a heterologous probe for the isolation of a n. crassa gene(designated chs-4) encoding a poly ...19968628221
transcriptional analysis of the mta idiomorph of neurospora crassa identifies two genes in addition to neurospora crassa, mating and heterokaryon formation between opposite mating-types is controlled by a single locus with two alternate forms termed mt a and mt a. previously, an open reading frame (mt a-1) that confers mating identity and heterokaryon incompatibility was characterized in the 5.3 kb mt a idiomorph. in this study, we describe the structural and transcriptional characterization of two additional genes in the mt a idiomorph, mt a-2 and mt a-3. the 373 amino acid mt a-2 orf has 23% ...19968628238
characterization of the biophysical and motility properties of kinesin from the fungus neurospora crassa.neurospora kinesin (nkin) is a distant relative of the family of conventional kinesins, members of which have been identified in various animal species. as in its animal counterparts, nkin most likely is an organelle motor. because it is a functional homologue of the kinesin heavy chain of higher eukaryotes, its biophysical and motility properties were compared with those of other conventional kinesins. purified nkin behaves as a homodimeric complex composed of two subunits of a 105-kda polypept ...19968631782
role of an upstream open reading frame in mediating arginine-specific translational control in neurospora crassa.the neurospora crassa arg-2 transcript contains an upstream open reading frame (uorf) specifying a 24-residue leader peptide and is subject to a novel form of negative translational regulation in response to arginine. the role of the arg-2 uorf in arginine-specific negative regulation was investigated by using translational fusions of wild-type and mutant arg-2 sequences to the escherichia coli lacz reporter gene specifying beta-galactosidase. the wild-type uorf conferred arg-specific regulation ...19968636015
insights into the molecular basis of thermal stability from the structure determination of pyrococcus furiosus glutamate dehydrogenase.the structure determination of the glutamate dehydrogenase from the hyperthermophile pyrococcus furiosus has been completed at 2.2 a resolution. the structure has been compared with the glutamate dehydrogenases from the mesophiles clostridium symbiosum, escherichia coli and neurospora crassa. this comparison has revealed that the hyperthermophilic enzyme contains a striking series of networks of ion-pairs which are formed by regions of the protein which contain a high density of charged residues ...19968639325
the sequence of a small subunit of cytochrome c oxidase from crithidia fasciculata which is homologous to mammalian subunit iv.the sequence of subunit 8 of cytochrome c oxidase from crithidia fasciculata was determined by sequencing cdna and n-terminus of the mature protein (mr = 15.7 kda). the (inferred) protein is homologous to mammalian cox iv and the corresponding cox subunits from yeast, neurospora crassa and dictyostelium discoideum, which is reflected in a very similar hydropathy profile. elements that are conserved in the c. fasciculata sequence include (i) an n-terminal (d/e)-(k/r)-x-k-(x2)-w-(x2)-(i/l) motif, ...19968641419
signal transduction in pneumocystis carinii: characterization of the genes (pcg1) encoding the alpha subunit of the g protein (pcg1) of pneumocystis carinii carinii and pneumocystis carinii ratti.pneumocystis carinii is a eukaryotic organism that causes pneumonia in immunocompromised hosts. the cell biology and life cycle of the organism are poorly understood primarily because of the lack of a continuous in vitro cultivation system. these limitations have prevented investigation of the organism's infectious cycle and hindered the rational development of new antimicrobial therapies and implementation of measures to prevent exposure to the organism or transmission. the interaction of p. ca ...19968641768
a receptor for the import of proteins into human mitochondria.we have characterised a 16.3-kda human protein that functions as a receptor for the import of preproteins into mitochondria. based on amino acid sequence alignments, the protein (hmas20p) is 41% similar to mas20p (20-kda mitochondrial assembly protein) from yeast saccharomyces cerevisiae and 38% similar to mom19 (19-kda mitochondrial outer-membrane protein) from neurospora crassa. hmas20p has a putative n-terminal transmembrane sequence of 29 amino acids and an acidic c-terminus. a 13-kda fragme ...19968654425
isolation and sequence analysis of the cdna encoding delta 1-pyrroline-5-carboxylate reductase from zalerion arboricola.a cdna encoding delta 1-pyrroline-5-carboxylate reductase (p5cr) was isolated from the pneumocandin (pmo)-producing fungus, zalerion arboricola (za), by complementation of a p5cr-deficient mutant (pro3) of saccharomyces cerevisiae (sc). the cloned cdna was placed under control of the sc galactokinase (gal1) promoter and restored p5cr activity to the pro3 mutant. sequence analysis revealed that the za p5cr-encoding cdna encodes an approx. 35 kda protein with substantial amino acid (aa) identity t ...19968654976
the regulatory protein nit4 that mediates nitrate induction in neurospora crassa contains a complex tripartite activation domain with a novel leucine-rich, acidic motif.expression of nit-3 and nit-6, the structural genes which encode nitrate reductase and nitrite reductase in neurospora crassa, requires the global-acting nit2 and the pathway specific nit4 regulatory proteins. nit4, which consists of 1090 amino-acid residues, possesses a cys6/zn2 zinc cluster dna-binding-domain. nit4 was dissected to localize transactivation domains by fusion of various segments of nit4 to the dna-binding domain of gal4 for in vivo analysis in yeast. three separate activation su ...19968662193
over-expression of the nud1-coded endo-exonuclease in saccharomyces cerevisiae enhances dna recombination and repair.the nud1(=nuc2) gene of saccharomyces cerevisiae has been subcloned and over-expressed in multi-copy plasmids. enhanced expression of this nuclear endo-exonuclease gene was confirmed by northern hybridization (>10-fold increase), and increased enzymatic activity (2.4-fold increase) was demonstrated by direct immunological assay using antibody raised against the purified neurospora crassa endo-exonuclease. we found that increased expression of nud1 was associated with an increase in cell survival ...19968662209
molecular cloning of thi-4, a gene necessary for the biosynthesis of thiamine in neurospora crassa.the thiamine-4 (thi-4) gene was cloned by functional complementation of a thi-4 mutant of neurospora crassa. the product of this gene is believed to be involved in the condensation of pyrimidine and thiazole precursors, which is necessary for the synthesis of thiamine. the thi-4 gene has ten introns which vary in length from 57 to 200 bp and the junction and internal (lariat) sequences are in good agreement with the consensus sequences for the splicing of introns in n. crassa. the thi-4 gene enc ...19968662211
characterization of the aspergillus parasiticus niad and niia gene cluster.the nitrate reductase gene (niad) and nitrite reductase gene (niia) of aspergillus parasiticus are clustered and are divergently transcribed from a 1.6-kb intergenic region (niad-niia). the deduced aminoacid sequence of the a. parasiticus nitrate reductase demonstrated a high degree of homology to those of other aspergillus species, as well as to leptosphaeria maculans, fusarium oxysporum, gibberella fujikuroi and neurospora crassa, particularly in the cofactor-binding domains for molybdenum, he ...19968662212
the role of the n and c termini of recombinant neurospora mitochondrial porin in channel formation and voltage-dependent investigate the role of the n and c termini in channel function and voltage-dependent gating of mitochondrial porin, we expressed wild-type and mutant porins from neurospora crassa as his-tag fusion products in escherichia coli. large quantities of the proteins were purified by chromatography across a nickle-nitrilotriacetic acid-agarose column under denaturing conditions. the purified his-tagged wild-type protein could be functionally reconstituted in the presence of detergent and sterol and ...19968662769
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