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secondary structure of the intact h+,k+-atpase and of its membrane-embedded region. an attenuated total reflection infrared spectroscopy, circular dichroism and raman spectroscopy study.models of p-type atpase predict that membrane-embedded fragments represent about 20% of the protein and adopt an all-alpha-helical structure. while this prediction was confirmed for the ca2-atpase [corbalan-garcia, s., teruel, j., villalain, j. & gomez-fernandez, j. (1994) biochemistry 33, 8247-8254], it is at odds with recent experimental evidence gathered on the neurospora crassa plasma membrane h+-atpase [vigneron, l., ruysschaert, j.-m. & goormaghtigh, e. (1995) j. biol. chem. 270, 17685-176 ...19989523697
molecular genetics of mating recognition in basidiomycete fungi.the recognition of compatible mating partners in the basidiomycete fungi requires the coordinated activities of two gene complexes defined as the mating-type genes. one complex encodes members of the homeobox family of transcription factors, which heterodimerize on mating to generate an active transcription regulator. the other complex encodes peptide pheromones and 7-transmembrane receptors that permit intercellular signalling. remarkably, a single species may have many thousands of cross-compa ...19989529887
characterization of the aspergillus nidulans nmra gene involved in nitrogen metabolite repression.the gene nmra of aspergillus nidulans has been isolated and found to be a homolog of the neurospora crassa gene nmr-1, involved in nitrogen metabolite repression. deletion of nmra results in partial derepression of activities subject to nitrogen repression similar to phenotypes observed for certain mutations in the positively acting area gene.19989537404
the neurospora rca-1 gene complements an aspergillus flbd sporulation mutant but has no identifiable role in neurospora sporulation.the aspergillus nidulans flbd gene encodes a protein with a myb-like dna-binding domain that is proposed to act in concert with other developmental regulators to control initiation of conidiophore development. we have identified a neurospora crassa gene called rca-1 (regulator of conidiation in aspergillus) based on its sequence similarity to flbd. we found that n. crassa rca-1 can complement the conidiation defect of an a. nidulans flbd mutant and that induced expression of rca-1 caused conidia ...19989539422
molecular analysis of a glucose-regulated gene (grp78) of neurospora crassa.the nucleotide sequence of the gene encoding the glucose-regulated protein 78 (grp78) of neurospora crassa was determined. the orf codes for a protein of 662 amino acids (72 kda) and belongs to the heat shock protein 70 (hsp70) gene family, which is characterized by three hsp70 'signature sequences'. the grp78 gene contains 5 introns. the protein carries the er retention signal hdel at its carboxy terminus and is most homologous to the kar2/grp78 protein of saccharomyces cerevisiae (78%) and to ...19989545520
oxidation of catalase by singlet oxygen.different bands of catalase activity in zymograms (cat-1a-cat-1e) appear during neurospora crassa development and under stress conditions. here we demonstrate that singlet oxygen modifies cat-1a, giving rise to a sequential shift in electrophoretic mobility, similar to the one observed in vivo. purified cat-1a was modified with singlet oxygen generated from a photosensitization reaction; even when the reaction was separated from the enzyme by an air barrier, a condition in which only singlet oxy ...19989553125
involvement of two novel chaperones in the assembly of mitochondrial nadh:ubiquinone oxidoreductase (complex i).the respiratory complex i of mitochondria consists of some 40 different subunits which form an l-shaped structure. perpendicular to a hydrophobic arm embedded in the inner mitochondrial membrane a peripheral arm protrudes into the matrix. assembly of the complex as studied in the fungus neurospora crassa involves the formation of discrete intermediates. the matrix arm and the membrane arm are formed independently of each other and are joined in the course of assembly. the membrane arm itself is ...19989769214
pzl-1 encodes a novel protein phosphatase-z-like ser/thr protein phosphatase in neurospora crassa.the gene and cdna of a novel protein phosphatase were cloned from neurospora crassa. the pzl-1 gene encompasses three introns and is localized to the left arm of chromosome i between cyt-21 and fsr-12. it encodes a protein of 58.3 kda containing a ser/pro rich n-terminal segment, and a c-terminal domain that is similar to the catalytic subunit of type 1 protein phosphatases. the first 51 amino acid residues, including a potential n-myristoylation site, as well as the c-terminal domain (about 300 ...19989774742
auto-adp-ribosylation of nad glycohydrolase from neurospora crassa.nad glycohydrolase (nadase; ec 3.2.2.5) is an enzyme that catalyzes hydrolysis of nad to produce adp-ribose and nicotinamide. we recently demonstrated that self-inactivation of nadase from rabbit erythrocytes was due to an auto-adp-ribosylation. in the present study, a mechanism of self-inactivation of nadase from neurospora crassa by its substrate was investigated by using intact mycelia of n. crassa and purified nadase, which had molecular characteristics different from mammalian nadases. the ...19989787786
isolation and characterization of a new gene, sre, which encodes a gata-type regulatory protein that controls iron transport in neurospora crassa.multiple gata factors - regulatory proteins with consensus zinc finger motifs that bind to dna elements containing a gata core sequence - exist in the filamentous fungus neurospora crassa. one gata factor, nit2. controls nitrogen metabolism, whereas two others, wc-1 and wc-2, regulate genes responsive to blue light induction. a gene encoding a new gata factor, named sre, was isolated from neurospora using a pcr-mediated method. sequence analysis of the new gata factor gene revealed an orf specif ...19989790585
identification of the first fungal annexin: analysis of annexin gene duplications and implications for eukaryotic evolution.annexin homologues have been found in animals, plants, and distinct protist lineages. we report the identification of the first fungal annexin, encoded by the anx14 gene of the filamentous ascomycete neurospora crassa. annexins have a complex evolutionary history and exhibit a large number of gene duplications and gene losses in various taxa, including the complete loss of annexin sequences from another ascomycete, the budding yeast saccharomyces cerevisiae. surprisingly, the n. crassa annexin h ...19989797403
the two-component hybrid kinase regulator canik1 of candida albicans.using degenerate primers of highly conserved regions of two-component response regulators for pcr amplification, a two-component response regulator was cloned from candida albicans that is homologous to nik-1+ of neurospora crassa. this two-component hybrid kinase, canik1, also shows features of bacterial two-component response regulators, including a putative unorthodox second histidine kinase motif at the carboxy-terminal end. canik1 was expressed at low levels in both the white and opaque swi ...19989802013
a new uv-sensitive mutant that suggests a second excision repair pathway in neurospora crassa.in an attempt to understand the relationship between photorepair and dark repair in neurospora crassa, a new mutant was isolated, which showed defects in both repair processes. the new mutant, mus-38, is moderately sensitive to uv and shows imperfect photoreactivation following uv irradiation. dna was purified from this mutant and the other uv-sensitive mutants, and analyzed for the removal of cyclobutane pyrimidine dimers (cpds). uv-specific endonuclease-sensitive sites (ess) completely disappe ...19989806416
characterization of aspergillus nidulans peroxisomes by immunoelectron microscopy.in previous work, we have demonstrated that oleate induces a massive proliferation of microbodies (peroxisomes) in aspergillus nidulans. although at a lower level, proliferation of peroxisomes also occurs in cells growing under conditions that induce penicillin biosynthesis. here, microbodies in oleate-grown a. nidulans cells were characterized by using several antibodies that recognize peroxisomal enzymes and peroxins in a broad spectrum of eukaryotic organisms such as yeast, and plant, and mam ...19989818356
analysis of actin and actin-related protein 3 (arp3) gene expression following induction of hyphal tip formation and apolar growth in neurospora.the genes encoding actin and arp3 in the filamentous fungus neurospora crassa were cloned and sequenced. the actin structural gene is interrupted by four introns and encodes a polypeptide of 375 amino acids, which shows very high degree of identity with actin from other sources. n. crassa arp3 is 439 amino acids in length and is 71% to 80% identical to arp3s from five other organisms, while actin is 40% to 50% identical to these same arp3s. transcript levels for actin and arp3 decrease upon indu ...19989819052
ssp1, a site-specific parvulin homolog from neurospora crassa active in protein folding.peptidyl-prolyl cis-trans-isomerases (ppiases) are enzymes capable of isomerizing a xaa-pro peptide bond. three families of ppiases are known: cyclophilins, fkbps, and parvulins. the physiological functions of the ppiases are only poorly understood. eucaryotic members of the parvulin family have recently been shown to be essential for regulation of mitosis. here we describe the purification and characterization of ssp1, an abundant parvulin homolog from neurospora crassa, which is unique among t ...19989822668
isolation and characterization of alpha-tubulin genes from septoria tritici and rhynchosporium secalis, and comparative analysis of fungal alpha-tubulin sequences.the alpha-tubulin genes from septoria tritici and rhynchosporium secalis have been cloned and sequenced. the predicted amino acid sequence and intron structure showed strong homology with other known filamentous fungal alpha-tubulins. comparison of sixteen fungal alpha-tubulin sequences based on amino acid sequence homology and intron structure identified five groups of proteins. group 1 consists of filamentous fungi, including s. tritici and r. secalis, the dimorphic fungus histoplasma capsulat ...19989829778
characterization of a gene encoding a single-subunit bacteriophage-type rna polymerase from maize which is alternatively spliced.single-subunit rna polymerases belonging to the t3/t7 bacteriophage family are thought to be common throughout eukaryotes. we report the isolation and characterization of a nucleus-encoded single-subunit rna polymerase gene from maize. this gene is highly homologous to other single-subunit rna polymerase genes from arabidopsis, chenopodium. yeast and neurospora crassa involved in organellar transcription. genomic southern analysis reveals 10 to 15 hybridising fragments, suggesting that maize con ...19989829825
phosphate transporter gene family of arabidopsis thaliana.using a high-affinity phosphate transporter gene of arabidopsis thaliana, pht1, as a probe, three arabidopsis homologs were isolated, named pht4, pht5 and pht6, in addition to the previously isolated pht2 and pht3. the amino acid sequences deduced from the three nucleotides were 32-42% homologous with microbial phosphate transporters of saccharomyces cerevisiae (pho84), neurospora crassa (pho-5) and glomus versiforme (gvpt). pht1, pht2, pht3 and pht6 were clustered in a 25-kbp region of chromoso ...19989872450
molybdenum-free nitrate reductases from vanadate-reducing bacteria.two catalytically distinct molybdenum-free dissimilatory nitrate reductases, a soluble periplasmic and a membrane-bound one, were isolated from the vanadate-reducing facultatively anaerobic bacterium pseudomonas isachenkovii and purified to electrophoretic homogeneity. the enzymes did not contain molybdenum, the periplasmic enzyme contained vanadium, whereas the membrane-bound enzyme was vanadium-free. both nitrate reductases lacked molybdenum cofactor. this fact was proved by reconstitution of ...19989883895
the fluffy gene of neurospora crassa encodes a gal4p-type c6 zinc cluster protein required for conidial development.neurospora crassa fluffy (fl) mutants are unable to produce macroconidia. we cloned the fl gene to determine its role in regulating conidiation. a cosmid clone containing fl was identified by complementation. the sequence of fl revealed that it encodes a gal4p-type c6 zinc cluster protein with greatest similarity to the n. crassa nit4 protein that regulates genes required for nitrate utilization. analysis of several fl mutant alleles demonstrated that null mutants are blocked in the budding phas ...19989560395
cloning and characterization of the yeast rad1 homolog gene (mus-38) from neurospora crassa: evidence for involvement in nucleotide excision repair.a neurospora crassa gene encoding a product with homology to the saccharomyces cerevisiae rad1 nucleotide excision repair (ner) protein was isolated by degenerate pcr. the predicted protein consists of 892 amino acids with a molecular weight of 100.4 kda, and 32-37% identity to the xpf/ercc4 protein family. the homolog was mapped to the left arm of linkage group i, the location of the mus-38 gene. subsequently, gene inactivation and complementation studies identified the rad1 homolog as mus-38. ...19989560435
three-dimensional structure of bovine nadh:ubiquinone oxidoreductase (complex i) at 22 a in ice.nadh:ubiquinone oxidoreductase (complex i) is the first and largest complex in the electron transport chain of mitochondria. the bovine complex purified from cardiac muscle consists of at least 42 different subunits with a combined molecular mass of about 890 kda. the three-dimensional structure of the complex was determined at 22 a from single particles embedded in vitrified ice using electron cryo-microscopy. the structure was calculated using a new program to align particles, to correct for t ...19989571020
repeat-induced point mutations in pad-1, a putative rna splicing factor from neurospora crassa, confer dominant lethal effects on ascus development.we describe the characterization of a gene, pad-1, from neurospora crassa which displays sequence characteristics of the rs class of hnrna-binding proteins (hnrnp) and mrna splicing factors. this is the first report of the isolation of a putative hnrnp gene from n. crassa. pad-1 showed 30% identity and 57% similarity to a protein, hcc1, which was isolated using autoantibodies from patients suffering from hepatocellular carcinoma. both hcc1 and pad-1 show amino acid sequence similarities to the h ...19989578630
role of fungal dynein in hyphal growth, microtubule organization, spindle pole body motility and nuclear migration.cytoplasmic dynein is a microtubule-associated motor protein with several putative subcellular functions. sequencing of the gene (dhc1) for cytoplasmic dynein heavy chain of the filamentous ascomycete, nectria haematococca, revealed a 4,349-codon open reading frame (interrupted by two introns) with four highly conserved p-loop motifs, typical of cytoplasmic dynein heavy chains. the predicted amino acid sequence is 78.0% identical to the cytoplasmic dynein heavy chain of neurospora crassa, 70.2% ...19989580563
the isolation and characterization of nrc-1 and nrc-2, two genes encoding protein kinases that control growth and development in neurospora crassa.using an insertional mutagenesis approach, a series of neurospora crassa mutants affected in the ability to control entry into the conidiation developmental program were isolated. one such mutant, gth16-t4, was found to lack normal vegetative hyphae and to undergo constitutive conidiation. the affected gene has been named nrc-1, for nonrepressible conidiation gene #1. the nrc-1 gene was cloned from the mutant genomic dna by plasmid rescue, and was found to encode a protein closely related to the ...19989584090
role of the negative charges in the cytosolic domain of tom22 in the import of precursor proteins into mitochondria.tom22 is an essential mitochondrial outer membrane protein required for the import of precursor proteins into the organelles. the amino-terminal 84 amino acids of tom22 extend into the cytosol and include 19 negatively and 6 positively charged residues. this region of the protein is thought to interact with positively charged presequences on mitochondrial preproteins, presumably via electrostatic interactions. we constructed a series of mutant derivatives of tom22 in which 2 to 15 of the negativ ...19989584158
cell wall active antifungal compounds produced by the marine fungus hypoxylon oceanicum ll-15g256. iii. biological properties of 15g256 gamma.15g256 gamma is a cyclic lipopeptide antifungal agent discovered in a mechanism of action screen for cell wall acting antifungal agents. the compound shows moderate activity in both greenhouse tests against plant disease caused by pathogenic fungi and in in vitro tests against human fungal pathogens. microscopic examination of treated fungi suggests that the compound acts by the inhibition of cell wall biosynthesis. however, in vitro inhibition of neurospora crassa glucan and chitin synthase wer ...19989589067
bacterial expression and characterization of the mitochondrial outer membrane channel. effects of n-terminal modifications.several forms of the voltage-dependent anion-selective channel (vdac) have been expressed at high yield in escherichia coli. full-length constructs of the proteins of neurospora crassa and saccharomyces cerevisiae (ncvdac and scvdac) have been made with 20-residue-long, thrombin-cleavable, his6-containing n-terminal extensions. ncvdac purified from bacteria or mitochondria displays a far-uv cd spectrum (in 1% lauryl dimethylamine oxide at ph 6-8) similar to that of bacterial porins, indicating e ...19989593723
complex i from the fungus neurospora crassa.respiratory chain complex i is a complicated enzyme of mitochondria, that couples electron transfer from nadh to ubiquinone to the proton translocation across the inner membrane of the organelle. the fungus neurospora crassa has been used as one of the main model organisms to study this enzyme. complex i is composed of multiple polypeptide subunits of dual genetic origin and contains several prosthetic groups involved in its activity. most subunits have been cloned and those binding redox centre ...19989593837
transcriptional activation of the aspergillus nidulans gpda promoter by osmotic signals.a differentially expressed gpda cdna clone was isolated from nacl-adapted aspergillus nidulans (fgsc359) and identified as glyceraldehyde-3-phosphate dehydrogenase (gpda) on the basis of its nucleotide sequence. the level of gpda rna substantially increased in cultures gradually adapted to nacl but was greatly reduced in cultures exposed briefly to a high concentration of nacl. a pyrg auxotroph of a. nidulans (a773) was cotransformed with a gpda-uida construct and a plasmid containing the neuros ...19989603839
cloning and sequencing of a cdna encoding a copper-zinc superoxide dismutase enzyme from the marine yeast debaryomyces hansenii.cu-zn superoxide dismutase (sod-1) is a ubiquitously occurring eukaryotic enzyme with a variety of important effects on respiring organisms. a gene (dhsod-1) encoding a cu-zn superoxide dismutase of the marine yeast debaryomyces hansenii was cloned using mrna by the rt-pcr technique. the deduced amino-acid sequence shows approximately 70% homology with that of cytosolic superoxide dismutase from saccharomyces cerevisiae and neurospora crassa, as well as lower homologies (between 55 and 65%) with ...19989605507
molecular characterisation of the 76 kda iron-sulphur protein subunit of potato mitochondrial complex i.genes encoding subunits of complex i (ec 1.6.5.3) of the mitochondrial respiratory chain vary in their locations between the mitochondrial and nuclear genomes in different organisms, whereas genes for a homologous multisubunit complex in chloroplasts have to date only been found on the plastid genome. in potato (solanum tuberosum l.), the gene coding for the mitochondrial 76 kda iron-sulphur protein is identified in the nuclear genome. the gene is transcribed into polyadenylated mrna which is mo ...19989615461
cos1, a two-component histidine kinase that is involved in hyphal development in the opportunistic pathogen candida albicans.two-component histidine kinases recently have been found in eukaryotic organisms including fungi, slime molds, and plants. we describe the identification of a gene, cos1, from the opportunistic pathogen candida albicans by using a pcr-based screening strategy. the sequence of cos1 indicates that it encodes a homolog of the histidine kinase nik-1 from the filamentous fungus neurospora crassa. cos1 is also identical to a gene called canik1 identified in c. albicans by low stringency hybridization ...19989618540
the sensor regions of vdac are translocated from within the membrane to the surface during the gating processes.the motion of the sensor regions in a mitochondrial voltage-gated channel called vdac were probed by attaching biotin at specific locations and determining its ability to bind to added streptavidin. site-directed mutagenesis was used to introduce single cysteine residues into neurospora crassa vdac (naturally lacks cysteine). these were chemically biotinylated and reconstituted into planar phospholipid membranes. in the 19 sites examined, only two types of results were observed upon streptavidin ...19989635747
functional analysis of different regions of the positive-acting cys3 regulatory protein of neurospora crassa.in the filamentous fungus neurospora crassa during conditions of sulfur limitation, cys3, a major positive-acting regulatory protein, turns on the expression of an entire set of genes which encode permeases and enzymes involved in the acquisition of sulfur from environmental sources. cys3 functions as a homodimeric protein and possesses a b-zip domain that confers sequence-specific dna binding. expression of various hybrid gal4-cys3 fusion proteins in yeast was used to detect regions involved in ...19989644202
complementation of the mpg1 mutant phenotype in magnaporthe grisea reveals functional relationships between fungal hydrophobins.the functional relationship between fungal hydrophobins was studied by complementation analysis of an mpg1(-) gene disruption mutant in magnaporthe grisea. mpg1 encodes a hydrophobin required for full pathogenicity of the fungus, efficient elaboration of its infection structures and conidial rodlet protein production. seven heterologous hydrophobin genes were selected which play distinct roles in conidiogenesis, fruit body development, aerial hyphae formation and infection structure elaboration ...19989670001
identification, cloning and characterization of a derepressible na+-coupled phosphate transporter in saccharomyces cerevisiae.based on the high sequence homology between the yeast orf ybr296c (accession number p38361 in the swiss-prot database) and the pho4 gene of neurospora crassa, which codes for a na+/pi cotransporter with twelve putative transmembrane domains, the ybr296c orf was considered to be a promising candidate gene for a plasma membrane-bound phosphate transporter in saccharomyces cerevisiae. therefore, this gene, here designated pho89, was cloned and a set of deletion mutants was constructed. we then stud ...19989671031
the essential aspergillus nidulans gene pmaa encodes an homologue of fungal plasma membrane h(+)-atpases.pmaa, an aspergillus nidulans gene encoding a p-atpase, has been cloned by heterologous hybridization with the yeast pma1 gene. the putative 990-residue pmaa polypeptide shows 50% identity to saccharomyces cerevisiae and neurospora crassa plasma membrane h(+)-atpases and weak (19-26%) identity to other yeast p-type cation-translocating atpases. pmaa contains all catalytic domains characterizing h(+)-atpases. pmaa transcript levels are not regulated by pacc, the transcription factor mediating ph ...19989680959
cpc-3, the neurospora crassa homologue of yeast gcn2, encodes a polypeptide with juxtaposed eif2alpha kinase and histidyl-trna synthetase-related domains required for general amino acid control.based on characteristic amino acid sequences of kinases that phosphorylate the alpha subunit of eukaryotic translation initiation factor 2 (eif2alpha kinases), degenerate oligonucleotide primers were constructed and used to polymerase chain reaction-amplify from genomic dna of neurospora crassa a sequence encoding part of a putative protein kinase. with this sequence an open reading frame was identified encoding a predicted polypeptide with juxtaposed eif2alpha kinase and histidyl-trna synthetas ...19989685394
phosphate permeases of saccharomyces cerevisiae.the pho84 and pho89 genes of saccharomyces cerevisiae encode two high-affinity phosphate cotransporters of the plasma membrane. hydropathy analysis suggests a secondary structure arrangements of the proteins in 12 transmembrane domains. the derepressible pho84 and pho89 transporters appear to have characteristic similarities with the phosphate transporters of neurospora crassa. the pho84 protein catalyzes a proton-coupled phosphate transport at acidic ph, while the pho89 protein catalyzes a sodi ...19989693717
redox components and structure of the respiratory nadh:ubiquinone oxidoreductase (complex i).the proton-pumping nadh:ubiquinone oxidoreductase is the first complex in the respiratory chains of many purple bacteria and of mitochondria of most eucaryotes. the bacterial complex consists of 14 different subunits. the mitochondrial complex contains at least 29 additional proteins that do not directly participate in electron transfer and proton translocation. we analysed electron micrographs of isolated and negatively stained complex i particles from escherichia coli and neurospora crassa and ...19989693737
a novel pathway for cytochromes c biogenesis in chloroplasts.the cytochromes c are a useful model for the study of the pathways and mechanisms of assembly of the cofactor-containing components of energy transducing membranes. genetic analyses have identified proteins that are required for the assembly of c-type cytochromes in mitochondria, bacteria and chloroplasts. the components of the pathway operating in fungal and animal mitochondria, i.e. the cytochrome (cyt) c and c1 heme lyases in the intermembrane space, were identified over a decade ago through ...19989693743
distribution patterns of 104 kda stress-associated protein in rice.a 104 kda protein (sap 104) accumulates in rice seedlings in response to several abiotic stress conditions and immunological homologues of rice sap 104 have been detected in several monocot and dicot species, as also neurospora crassa, a fungus. in this report, we show that the amino acid sequence of a tryptic peptide generated from purified sap 104 bears significant homology with an atp-binding domain of the hsp 100 family proteins of arabidopsis thaliana and glycine max. it is further shown th ...19989700064
dynamics of the tom complex of mitochondria during binding and translocation of preproteins.translocation of preproteins across the mitochondrial outer membrane is mediated by the tom complex. this complex consists of receptor components for the initial contact with preproteins at the mitochondrial surface and membrane-embedded proteins which promote transport and form the translocation pore. in order to understand the interplay between the translocating preprotein and the constituents of the tom complex, we analyzed the dynamics of the tom complex of neurospora crassa and saccharomyce ...19989710610
structure of the chromosome vii centromere region in neurospora crassa: degenerate transposons and simple repeats.dna from the centromere region of linkage group (lg) vii of neurospora crassa was cloned previously from a yeast artificial chromosome library and was found to be atypical of neurospora dna in both composition (at rich) and complexity (repetitive). we have determined the dna sequence of a small portion (approximately 16.1 kb) of this region and have identified a cluster of three new retrotransposon-like elements as well as degenerate fragments from the 3' end of tad, a previously identified line ...19989710630
structure, function, regulation: experimental analysis in groups of non-excitable cells coupled via permeable junctions.this presentation reviews the functional properties of different permeable junctions (microplasmodesmata of filamentous cyanobacteria, septal pores of fungi, and gap junctions of animals). examples of the distribution of power by ionic fluxes and electric current through permeable cell-cell junctions are considered. based on the analysis of intercellular communication occurring during apical growth of hyphae of wild-type and mutant lines of the mycelial fungus neurospora crassa, approaches to st ...19989718577
in vitro studies on mode of action of antifungal 8.o.4'-neolignans occurring in certain species of virola and related genera of myristicaceae.neutral racemic antifungal alcohols of 8.o.4'-neolignan type, were evaluated for inhibitory activity towards the fungal cell wall, using the whole cell neurospora crassa hyphal growth inhibition assay. results strongly suggested that these compounds could act by inhibiting cell wall polymer synthesis or assembly. active compounds were tested for their inhibitory activities against (1,3)-beta-glucan synthase, an enzyme that catalyzes the synthesis of the major wall polymer (1,3)-beta-glucan. alth ...19989720609
pyrrolnitrin from burkholderia cepacia: antibiotic activity against fungi and novel activities against streptomycetes.a bacterial strain identified as burkholderia cepacia nb-1 was isolated from water ponds in the botanical garden in tübingen, germany, and was found to produce a broad spectrum phenylpyrrole antimicrobial substance active against filamentous fungi, yeasts and gram-positive bacteria. in batch culture containing glycerol and l-glutamic acid, the isolate nb-1 produced the antibiotic optimally late in the growth phase and accumulated a main portion in their cells. isolation and purification of the a ...19989721657
mapping fungal ion channel locationsion channel mapping techniques are described and the results for two fungal organisms, saprolegnia ferax and neurospora crassa, are presented. in these species, two channel types have been characterized, stretch-activated channels exhibiting significant calcium permeability and spontaneous channels having significant potassium permeability. two distinct analyses of patch clamp data, analysis of channel self-clustering and association between different channel types, and localization along the hy ...19989742193
agrobacterium tumefaciens-mediated transformation of filamentous fungi.agrobacterium tumefaciens transfers part of its ti plasmid, the t-dna, to plant cells during tumorigenesis. it is routinely used for the genetic modification of a wide range of plant species. we report that a. tumefaciens can also transfer its t-dna efficiently to the filamentous fungus aspergillus awamori, demonstrating dna transfer between a prokaryote and a filamentous fungus. we transformed both protoplasts and conidia with frequencies that were improved up to 600-fold as compared with conve ...19989743116
single mating type-specific genes and their 3' utrs control mating and fertility in cochliobolus heterostrophus.to determine the number of proteins required for mating type (mat) locus-regulated control of mating in cochliobolus heterostrophus, mat fragments of various sizes were expressed in mat deletion strains. as little as 1.5 kb of mat sequence, encoding a single unique protein in each mating type (mat-1 and mat-2), conferred mating ability, although an additional 160 bp of 3' utr was needed for production of ascospores. no other mating type-specific genes involved in mating identity or fertility wer ...19989749670
engineering of pyridine nucleotide specificity of nitrate reductase: mutagenesis of recombinant cytochrome b reductase fragment of neurospora crassa nadph:nitrate reductase.the cytochrome b reductase fragment of neurospora crassa nadph:nitrate reductase (ec 1.6.6.3) was overexpressed in escherichia coli with a his-tag for purification after mutation of the nadph binding site. the recombinant enzyme fragment was altered by site-directed mutagenesis guided by the three-dimensional structure of cytochrome b reductase fragment of corn nadh:nitrate reductase (ec 1.6.6.1). substitution of asp for ser920 (using residue numbering for holo-nadph:nitrate reductase of n. cras ...19989750171
cloning and characterization of two phosphate transporters from medicago truncatula roots: regulation in response to phosphate and to colonization by arbuscular mycorrhizal (am) fungi.most vascular plants can acquire phosphate from the environment either directly, via the roots, or indirectly, via a fungal symbiont that invades the cortical cells of the root. here we have identified two cdna clones (mtpt1 and mtpt2) encoding phosphate transporters from a mycorrhizal root cdna library (medicago truncatula/glomus versiforme). the cdnas represent m. truncatula genes and the encoded proteins share identity with high-affinity phosphate transporters from arabidopsis, potato, yeast, ...19989425684
synthesis and differential turnover of the cys3 regulatory protein of neurospora crassa are subject to sulfur control.the transcription factor cys3 of neurospora crassa is a positive regulator of the sulfur regulatory circuit which contains many structural genes involved in sulfur metabolism. expression and degradation of the cys3 protein are precisely regulated in a sulfur-dependent manner. cys-3 expression was found to be fully repressed by high concentrations of methionine or inorganic sulfate present in the culture medium and to be derepressed when these favored sulfur sources were limited. cys-3 transcript ...19989457847
recombinant pseudorabies virus dnase exhibits a recbcd-like catalytic function.the pseudorabies virus (prv) dnase gene has previously been mapped within the prv genome. to characterize further the enzymic properties of prv dnase, this enzyme was expressed in escherichia coli with the use of a pet expression vector. the protein was purified to homogeneity and assayed for nuclease activity in vitro. recombinant prv dnase exhibited an alkaline ph preference and an absolute requirement for mg2+ ions that could not be replaced by ca2+ and na+ ions. further studies showed that p ...19989461490
differential expression of glucose-regulated (grp78) and heat-shock-inducible (hsp70) genes during asexual development of neurospora crassa.the expression of a glucose-regulated gene (grp78) changes significantly during the vegetative life cycle of neurospora crassa: the amounts of grp78 mrna are low in dormant conidia, increase during germination and exponential growth, decline in young aerial hyphae and reach a maximum in late (15-18 h) aerial hyphae. heat shock (30 min at 45 degrees c) elevated the mrna level of this gene especially in early aerial hyphae, whereas no increase above the high constitutive amount was found after hea ...19989467899
the neurospora aab-1 gene encodes a ccaat binding protein homologous to yeast hap5.the expression of the am (glutamate dehydrogenase) gene is dependent upon two upstream activating sequences, designated ursam(alpha) and ursam(beta). a heteromeric nuclear protein am alpha binding protein (aab) binds specifically to a ccaat box within the ursam(alpha) element. aab appears to be composed of three components. we used polyclonal antiserum raised against the highly purified aab1 subunit to isolate a partial aab-1 cdna clone, which was then used to isolate a full-length cdna and a ge ...19989475726
isolation of a class iv chitin synthase gene from a zygomycete fungus, rhizopus oligosporus.we found the presence of dna sequence which shows sequence similarity to the class iv chitin synthase gene (chs3) of saccharomyces cerevisiae in the genome of 14 rhizopus species which belong to zygomycetes. we cloned a gene (chs3), which might correspond to one of these homologous sequences, from rhizopus oligosporus by low stringency plaque hybridization probed with chs3. the deduced amino acid sequence of this gene showed highest similarity to the class iv chitin synthase of neurospora crassa ...19989851030
isolation of a nitrogen response regulator gene (nrr1) from metarhizium anisopliae.attempts to improve the effectiveness of entomopathogenic fungi as biological control agents require a clear understanding of the pathogenicity determinants at both the biochemical and molecular level. proteases play a key role in entomopathogenicity, allowing the fungus to penetrate the insect cuticle and rapidly invade the host. the most extensively studied of these protease activities, pr1a and pr2, are both subject to nitrogen derepression. the metarhizium anisopliae nrr1 (nitrogen response ...19989852945
[arg-13 is implicated in mitochondrial ornithine transport in neurospora crassa].arg-13 is a leaky mutation involved in arginine metabolism. a tight selection is developed using similar amount of lysine and ornithine replacing other nitrogen source in minimal medium. this selection strongly inhibits the growth of arg-13 under stringent sorbose/glucose condition but allows arg-13 to grow under spot test conditions. as ornithine is build up through mitochondrial ornithine biosynthesis and transport from cytoplasm to mitochondria, arg-13 is combined in genetic crosses with arg- ...199812016957
comparison of chitin content in the apical and distal parts of fungal hyphae in basidiobolus ranarum, neurospora crassa and coprinus sterquilinus.primary cell wall is synthesized in the growth zone of hyphal apex in fungi and rigidified during maturation along the newly formed hypha. cross-linking of cell-wall components and self-assembly of individual polysaccharide chains into microfibrils are supposed to be involved in the rigidification process. we determined the relative chitin content in the cell wall of hyphal tips and distal walls of three fungal species and demonstrated a general increase in relative chitin content in mature cell ...199910983235
ancf, the ccaat binding complex of aspergillus nidulans, contains products of the hapb, hapc, and hape genes and is required for activation by the pathway-specific regulatory gene amdr.ccaat binding factors (cbfs) positively regulating the expression of the amds gene (encoding acetamidase) and two penicillin biosynthesis genes (ipna and aata) have been previously found in aspergillus nidulans. the factors were called ancf and penr1, respectively. deletion of the hapc gene, encoding a protein with significant similarity to hap3p of saccharomyces cerevisiae, eliminated both ancf and penr1 binding activities. we now report the isolation of the genes hapb and hape, which encode pr ...19999858535
similarity pattern analysis in mutational distributions.the validity and applicability of the statistical procedure - similarity pattern analysis (span) - to the study of mutational distributions (mds) was demonstrated with two sets of data. the first was mutational spectra (ms) for 697 gc to at transitions produced with eight alkylating agents (aas) in the laci gene of escherichia coli. the second was a recently summarized data on the distributions of 11562 spontaneous, radiation- and chemical-induced forward mutations in the ad-3 region of heteroka ...199910592318
disruption of the gene for hsp30, an alpha-crystallin-related heat shock protein of neurospora crassa, causes defects in import of proteins into mitochondria.the gene for hsp30, the only known alpha-crystallin-related heat shock protein of neurospora crassa, was disrupted by repeat-induced point mutagenesis, leading to loss of cell survival at high temperature. hsp30, which is not synthesized at 30 degrees c, associates reversibly with the mitochondria at high temperature (45 degrees c). in this study, we found that import of selected proteins into internal compartments of mitochondria, following their synthesis in the cytosol, was severely impaired ...199910595587
use of atp6 in fungal phylogenetics: an example from the boletales.complete nucleotide sequences have been determined for atp6 from suillus luteus and cox3 from suillus sinuspaulianus (boletales, hymenomycetes, basidiomycota), which code for atpase subunit 6 and cytochrome oxidase subunit 3, respectively. these sequences were used to design pcr primers for the amplification of partial atp6 and cox3 sequences from other members of the boletales and outgroup taxa. in atp6 and cox3 from russula rosacea, one of the outgroup taxa, we observed a number of in-frame tg ...199910620406
cloning and characterization of a gene (arpa) from aspergillus oryzae encoding an actin-related protein required for normal nuclear distribution and morphology of conidiophores.we have isolated the arpa gene from aspergillus oryzae as a homologue of the neurospora crassa ro-4 gene. in n. crassa, mutations in the ro-4 gene, which encodes a major component of the dynactin complex arp1, causes curling of hyphae and abnormalities in nuclear distribution. the arpa gene contains two introns and encodes a polypeptide of 381 amino acids, with a 78% sequence identity to the n. crassa arp1. overexpression of the arpa gene causes a defect in nuclear migration into elongating hyph ...199910628858
in vitro antifungal evaluation and studies on the mode of action of xanthoxyline derivatives.this study describes the fungistatic effect of xanthoxyline (cas 90-24-4) and its derivatives against a panel of yeasts, filamentous fungi and dermatophytes, by using the agar dilution method. results indicated that simple structural modifications led to more potent derivatives, especially in relation with dermatophytes. the most active compound tested (10), which is a benzenesulphonyl derivative, was 12-fold more potent than xanthoxyline itself against trichophyton rubrum. the evaluation of the ...199910635452
posttranscriptional gene silencing in neurospora by a recq dna helicase.the phenomenon of posttranscriptional gene silencing (ptgs), which occurs when a transgene is introduced into a cell, is poorly understood. here, the qde-3 gene, which is required for the activation and maintenance of gene silencing in the fungus neurospora crassa, was isolated. sequence analysis revealed that the qde-3 gene belongs to the recq dna helicase family. the qde3 protein may function in the dna-dna interaction between introduced transgenes or with an endogenous gene required for gene- ...199910600745
a highly conserved mechanism of regulated ribosome stalling mediated by fungal arginine attenuator peptides that appears independent of the charging status of arginyl-trnas.the arg attenuator peptide (aap) is an evolutionarily conserved peptide involved in arg-specific negative translational control. it is encoded as an upstream open reading frame (uorf) in fungal mrnas specifying the small subunit of arg-specific carbamoyl phosphate synthetase. we examined the functions of the saccharomyces cerevisiae cpa1 and neurospora crassa arg-2 aaps using translation extracts from s. cerevisiae, n. crassa, and wheat germ. synthetic rna containing aap and firefly luciferase ( ...199910608810
histidine kinase, two-component signal transduction proteins of candida albicans and the pathogenesis of candidosis.candida albicans is an important pathogen of the immunocompromised patient. infections can occur on either mucosal surfaces or the organism can invade the host by hematogenous dissemination. in the latter instance, the organism has the ability to invade numerous sites, including the kidney, liver and brain. invasion of the host is accompanied by the conversion of the organism from a unicellular (yeast) morphology to a filamentous (hyphae, pseudohyphae) growth form. the morphogenetic change which ...199910865904
chitin synthase genes in the arbuscular mycorrhizal fungus glomus versiforme: full sequence of a gene encoding a class iv chitin synthase.chitin synthase genes of the arbuscular mycorrhizal fungus glomus versiforme were sought in an investigation of the molecular basis of fungal growth. three dna fragments (gvchs1, gvchs2 and gvchs3) corresponding to the conserved regions of distinct chitin synthase (chs) genes were amplified by means of the polymerase chain reaction (pcr) with two sets of degenerate primers. gvchs1 and gvchs2 encode two class i chitin synthases, whereas gvchs3 encodes a class iv chitin synthase. a genomic library ...19999919652
cloning and expression of the cdna encoding an alternative oxidase gene from aspergillus niger wu-2223l.a cdna fragment encoding the mitochondrial alternative oxidase, the enzyme responsible for cyanide-insensitive and salicylhydroxamic acid (sham)-sensitive respiration, from the citric acid-producing fungus aspergillus niger wu-2223l was cloned and expressed in escherichia coli as a host strain. synthetic primers were designed from the conserved nucleotide sequences of the alternative oxidase genes from higher plants and a yeast. the 210-bp dna fragment was amplified by pcr with these primers usi ...19999933359
the aspergillus nidulans gata factor srea is involved in regulation of siderophore biosynthesis and control of iron uptake.a gene encoding a new gata factor from aspergillus nidulans, srea, was isolated and characterized. srea displays homology to two fungal regulators of siderophore biosynthesis: about 30% overall identity to sre from neurospora crassa and about 50% identity to urbs1 from ustilago maydis over a stretch of 200 amino acid residues containing two gata-type zinc finger motifs and a cysteine-rich region. this putative dna binding domain, expressed as a fusion protein in escherichia coli, specifically bi ...19999988696
cloning of two genes encoding potassium transporters in neurospora crassa and expression of the corresponding cdnas in saccharomyces cerevisiae.two neurospora crassa genes, trk-1 and hak-1, encode k+ transporters that show sequence similarities to the trk transporters described in saccharomyces cerevisiae and schizosaccharomyces pombe, and to the hak transporters described in schwanniomyces occidentalis and barley. the n. crassa trk1 and hak1 transporters expressed by the corresponding cdnas in a trk1 delta trk2 delta mutant of s. cerevisiae exhibited a high affinity for rb+ and k+. northern blot analysis and comparison of the kinetic c ...199910027968
the wd protein cpc2p is required for repression of gcn4 protein activity in yeast in the absence of amino-acid starvation.the cpc2 gene of the budding yeast saccharomyces cerevisiae encodes a g beta-like wd protein which is involved in regulating the activity of the general control activator gcn4p. the cpc2 gene encodes a premrna which is spliced and constitutively expressed in the presence or absence of amino acids. loss of cpc2 gene function suppresses a deletion of the gcn2 gene encoding the general control sensor kinase, but not a deletion in the gcn4 gene. the resulting phenotype has resistance against amino-a ...199910048025
isolation, characterization and disruption of the area nitrogen regulatory gene of gibberella fujikuroi.the gene area-gf, a homologue of the major nitrogen regulatory genes nit-2, area, nre and nut1 of neurospora crassa, aspergillus nidulans, penicillium chrysogenum and magnaporthe grisea, respectively, was cloned from the gibberellin (ga)-producing rice pathogen gibberella fujikuroi. area-gf encodes a protein of 972 amino acid residues which contains a single putative zinc finger dna-binding domain that is at least 98% identical to the zinc finger domains of the homologous fungal proteins. the ar ...199910071216
positive regulation of adenylyl cyclase activity by a galphai homolog in neurospora crassa.gna-1 and gna-2 are two g protein alpha subunits from the filamentous fungus neurospora crassa. loss of gna-1 leads to multiple phenotypes, while deltagna-2 strains do not exhibit visible defects. however, deltagna-1deltagna-2 mutants are more affected in deltagna-1 phenotypes. here we report a biochemical investigation of the roles of gna-1 and gna-2 in camp metabolism. assays of mg2+ atp-dependent adenylyl cyclase activity (+/-gppnhp) in extracts from submerged cultures indicated that deltagna ...199910072319
pyruvate decarboxylase filaments are associated with the cortical cytoskeleton of asci and spores over the sexual cycle of filamentous ascomycetes.we show that pyruvate decarboxylase (pdc) 8- to 10-nm-diameter filaments, first described in vegetative cells of neurospora crassa, are ubiquitously present in filamentous fungi. the cellular arrangement of these structures was examined over the entire sexual cycle of the ascomycetes n. crassa, n. tetraesperma, podospora anserina, and sordaria macrospora. pdc-filaments were found associated with the cortical microtubule array of asci and ascospores and absent from the vicinity of spindles and sp ...199910072321
amphotropic murine leukemia virus entry is determined by specific combinations of residues from receptor loops 2 and 4.pit2 is the human receptor for amphotropic murine leukemia virus (a-mulv); the related human protein pit1 does not support a-mulv entry. interestingly, chimeric proteins in which either the n-terminal or the c-terminal part of pit2 was replaced by the pit1 sequence all retained a-mulv receptor function. a possible interpretation of these observations is that pit1 harbors sequences which can specify a-mulv receptor function when presented in a protein context other than pit1, e.g., in pit1-pit2 h ...199910074169
physiological consequence of disruption of the vma1 gene in the riboflavin overproducer ashbya gossypii.the vacuolar atpase subunit a structural gene vma1 of the biotechnologically important riboflavin overproducer ashbya gossypii was cloned and disrupted to prevent riboflavin retention in the vacuolar compartment and to redirect the riboflavin flux into the medium. cloning was achieved by polymerase chain reaction using oligonucleotide primers derived form conserved sequences of the vma1 proteins from yeast and filamentous fungi. the deduced polypeptide comprises 617 amino acids with a calculated ...199910092625
the ukc1 gene encodes a protein kinase involved in morphogenesis, pathogenicity and pigment formation in ustilago maydis.the fungal phytopathogen ustilago maydis alternates between budding and filamentous growth during its life cycle. this dimorphic transition, which is influenced by environmental factors and mating, is regulated in part by camp-dependent protein kinase (pka). we have recently identified a related protein kinase, encoded by the ukc1 gene, that also plays a role in determining cell shape. the ukc1 gene is homologous to several other protein kinase-encoding genes including the cot-i gene of neurospo ...199910102363
characterisation of the last fe-s cluster-binding subunit of neurospora crassa complex i.we have cloned cdnas encoding the last iron-sulphur protein of complex i from neurospora crassa. the cdna sequence contains an open reading frame that codes for a precursor polypeptide of 226 amino acid residues with a molecular mass of 24972 da. our results indicate that the mature protein belongs probably to the peripheral arm of complex i and is rather unstable when not assembled into the enzyme. the protein is highly homologous to the psst subunit of bovine complex i, the most likely candida ...199910216160
copper- and zinc-containing superoxide dismutase and its gene from candida albicans.cytosolic copper- and zinc-containing superoxide dismutase was purified 136-fold with an overall yield of 2.5% to apparent electrophoretic homogeneity from the dimorphic pathogenic fungus, candida albicans. the molecular mass of the native enzyme was 39.4 kda and the enzyme was composed of two identical subunits with a molecular mass of 19.6 kda. the enzyme was stable in the range of ph 4.0-9.0 and up to 55 degrees c. the ultraviolet-visible absorption spectrum of the enzyme showed the absorptio ...199910216241
a common ancestor for a subunit in the mitochondrial proton-translocating nadh:ubiquinone oxidoreductase (complex i) and short-chain dehydrogenases/reductases.the proton-translocating nadh:ubiquinone oxidoreductase or complex i is located in the inner membranes of mitochondria, where it catalyzes the transfer of electrons from nadh to ubiquinone. here we report that one of the subunits in complex i is homologous to short-chain dehydrogenases and reductases, a family of enzymes with diverse activities that include metabolizing steroids, prostaglandins and nucleotide sugars. we discovered that a subunit of complex i in human, cow, neurospora crassa and ...199910228558
neutral trehalases catalyse intracellular trehalose breakdown in the filamentous fungi aspergillus nidulans and neurospora crassa.a camp-activatable ca2+-dependent neutral trehalase was identified in germinating conidia of aspergillus nidulans and neurospora crassa. using a pcr approach, a. nidulans and n. crassa genes encoding homologues of the neutral trehalases found in several yeasts were cloned and sequenced. disruption of the antreb gene encoding a. nidulans neutral trehalase revealed that it is responsible for intracellular trehalose mobilization at the onset of conidial germination, and that this phenomenon is part ...199910320571
nad+-dependent glutamate dehydrogenase of the edible mushroom agaricus bisporus: biochemical and molecular characterization.the nad+-dependent glutamate dehydrogenase (nad-gdh) of agaricus bisporus, a key enzyme in nitrogen metabolism, was purified to homogeneity. the apparent molecular mass of the native enzyme is 474 kda comprising four subunits of 116 kda. the isoelectric point of the enzyme is about 7.0. km values for ammonium, 2-oxoglutarate, nadh, glutamate and nad+ were 6.5, 3.5, 0.06, 37.1 and 0.046 mm, respectively. the enzyme is specific for nad(h). the gene encoding this enzyme (gdhb) was isolated from an ...199910323225
interaction of mitochondrial presequences with dnak and mitochondrial hsp70.mitochondrial heat shock protein 70 (mt-hsp70) functions as a molecular chaperone in mitochondrial biogenesis. the chaperone in co-operation with its co-proteins acts as a translocation motor pulling the mitochondrial precursor into the matrix. mt-hsp70s are highly conserved when compared to the bacterial hsp70 homologue, dnak. here we have used dnak as a model to study the interaction of mitochondrial presequences with mt-hsp70 applying a dnak-binding algorithm, computer modeling and biochemica ...199910329135
signal presequences increase mitochondrial permeability and open the multiple conductance channel.we have reported that the signal presequence of cytochrome oxidase subunit iv from neurospora crassa increases the permeability of isolated rat liver mitochondria [p. m. sokolove and k. w. kinnally (1996) arch. biochem. biophys. 336, 69] and regulates the behavior of the mutiple conductance channel (mcc) of yeast inner mitochondrial membrane [t. a. lohret and k. w. kinnally (1995) j. biol. chem. 270, 15950]. here we examine in greater detail the action of a number of mitochondrial presequences f ...199910334870
molecular cloning and characterization of ct-pkar, a gene encoding the regulatory subunit of camp-dependent protein kinase in colletotrichum trifolii.colletotrichum trifolii is a plant pathogenic fungus causing alfalfa anthracnose. prepenetration development, including conidial germination and appressorial formation, are requisite for successful infection. pharmacological data from our laboratory indicated a role for a camp-dependent protein kinase (pka) pathway during these early morphogenic transitions. thus, the cloning and characterization of the genes for pka catalytic and regulatory subunits were undertaken to more precisely determine t ...199910339807
molecular mechanisms of chromosomal rearrangement in fungi.both sexual and asexual fungi undergo chromosomal rearrangements, which are the main cause of karyotype variability among the populations. different recombination processes can produce chromosomal reorganizations, both during mitosis and meiosis, but other mechanisms operate to limit the extent of the rearrangements; some of these mechanisms, such as the rip (repeat-induced point mutations) of neurospora crassa, have been well established for sexual fungi. in laboratory strains, treatments such ...199910342097
carbon regulation of ribosomal genes in neurospora crassa occurs by a mechanism which does not require cre-1, the homologue of the aspergillus carbon catabolite repressor, crea.transcription of the ribosomal protein and 40s rrna genes is coordinately regulated during steady state growth and carbon shifts in neurospora crassa. recognition sequences for the aspergillus nidulans carbon catabolite repressor, crea, overlap transcriptional elements of a 40s rrna gene and the crp-2 ribosomal protein gene. they also occur in similar locations in the promoters of several other ribosomal protein genes. substitutions encompassing the -74 and -167 crea consensus sequences in the c ...199910361038
identification of a 24 kda intrinsic membrane protein from mammalian peroxisomes.a 24 kda protein from rat liver peroxisomal membrane was isolated and subjected to edman degradation. using the n-terminal sequence of this polypeptide we have identified several rat and human expressed sequence tags in the genbank database. the complete sequence of a human cdna clone was determined. the open reading frame encodes an extremely basic protein 212 amino acid residues long. a high similarity between this mammalian protein and hypothetical proteins from caenorhabditis elegans and neu ...199910366717
hyperornithinaemia-hyperammonaemia-homocitrullinuria syndrome is caused by mutations in a gene encoding a mitochondrial ornithine transporter.neurospora crassa arg13 and saccharomyces cerevisiae arg11 encode mitochondrial carrier family (mcf) proteins that transport ornithine across the mitochondrial inner membrane. we used their sequences to identify est candidates that partially encode orthologous mammalian transporters. we thereby identified such a gene (ornt1) that maps to 13q14 and whose expression, similar to that of other urea cycle (uc) components, was high in liver and varied with changes in dietary protein. ornt1 expression ...199910369256
pagrg1, a glucose-repressible gene of podospora anserina that is differentially expressed during lifespan.a loss-of-function mutation in grisea, a gene of podospora anserina that was previously shown to code for a copper-modulated transcription factor, leads to a significant increase in lifespan. in an attempt to identify and to isolate potential target genes controlled by grisea, a rt-differential-display screen was performed. this approach resulted in the identification of a gene, pagrg1, that is differentially expressed in the wild-type and in the long-lived grisea mutant. in the mutant, transcri ...199910369964
a single external enzyme confers alternative nadh:ubiquinone oxidoreductase activity in yarrowia lipolytica.nadh:ubiquinone oxidoreductases catalyse the first step within the diverse pathways of mitochondrial nadh oxidation. in addition to the energy-conserving form commonly called complex i, fungi and plants contain much simpler alternative nadh:ubiquinone oxido-reductases that catalyze the same reaction but do not translocate protons across the inner mitochondrial membrane. little is known about the distribution and function of these enzymes. we have identified ylndh2 as the only gene encoding an al ...199910381390
assaying rna chaperone activity in vivo using a novel rna folding trap.in the absence of proteins, rnas often misfold in vitro due to alternative base pairings which result from the molecule being trapped in inactive conformations. we identify an in vivo folding trap in the t4 phage td gene, caused by nine base pairs between a sequence element in the upstream exon of the td gene and another at the 3' end of the intron. during translation, the ribosome resolves this interaction; consequently the intron folds correctly and splicing occurs. the introduction of a stop ...199910393192
transcription of the avirulence gene avr9 of the fungal tomato pathogen cladosporium fulvum is regulated by a gata-type transcription factor in aspergillus nidulans.the avirulence gene avr9 of the fungal tomato pathogen cladosporium fulvum is highly induced during infection of tomato plants. expression of the avr9 gene can also be induced in vitro when cells are grown on synthetic liquid medium containing little or no nitrogen. the avr9 promoter contains six copies of the sequence tagata and six additional copies of the core sequence gata within 0.4 kb upstream of the translation start site. in the filamentous fungi aspergillus nidulans and neurospora crass ...199910394902
comparison of h+-atpase and ca2+-atpase suggests that a large conformational change initiates p-type ion pump reaction cycles.structures have recently been solved at 8 a resolution for both ca2+-atpase from rabbit sarcoplasmic reticulum and h+-atpase from neurospora crassa. these cation pumps are two distantly related members of the family of p-type atpases, which are thought to use similar mechanisms to generate atp-dependent ion gradients across a variety of cellular membranes. we have undertaken a detailed comparison of the two structures in order to describe their similarities and differences as they bear on their ...199910395538
singlet oxygen is part of a hyperoxidant state generated during spore germination.we show that singlet oxygen is generated in asexual spores (conidia) from neurospora crassa at the onset of germination. oxidation of n. crassa catalase-1 (cat-1) was previously shown to be caused by singlet oxygen (lledías et al. j. biol. chem. 273, 1998). in germinating conidia, increased protein oxidation, decrease of total protein, cat-1 oxidation and accumulation of cat-1 mrna was detected. these changes were modulated in vivo by light intensity, an external clean source of singlet oxygen, ...199910401603
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