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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
the enzymic synthesis of beta-[32p]udp-n-acetylglucosamine.cells of micrococcus sp. 2102 incorporate inorganic [32p]phosphate from the medium into the sugar-phosphate polymer of the wall. controlled acid hydrolysis of sodium dodecyl sulphate-extracted cells gives n-acetylglucosamine 6-[32p]-phosphate which can be purified by ion-exchange chromatography and incubated with utp in the presence of crude preparations of phosphoacetylglucosamine mutase from neurospora crassa and utp:n-acetylglucosamine 1-phosphate phosphotransferase from bacillus licheniformi ...19789762094
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
mitop: database for mitochondria-related proteins, genes and diseases.the mitop database http://websvr.mips.biochem.mpg. de/proj/medgen/mitop/ consolidates information on both nuclear- and mitochondrial-encoded genes and their proteins. the five species files- saccharomyces cerevisiae, mus musculus, caenorhabditis elegans, neurospora crassa and homo sapiens -include annotated data derived from a variety of online resources and the literature. a wide spectrum of search facilities is given in the interelated sections 'gene catalogues', 'protein catalogues', 'homolog ...19999847163
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
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
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
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
flocculation of hyphae is associated with a deletion in the putative cahk1 two-component histidine kinase gene from candida albicans.in candida albicans, three putative histidine kinase genes have been described thus far, including casln1, canik1/cos1 and cahk1. the encoded proteins for c. albicans, casln1p and canik1p, which are similar to sln1p from saccharomyces cerevisiae and nik-1 from neurospora crassa, seem to function in osmoregulation and morphogenesis, respectively. recently, the isolation of cahk1, a putative histidine kinase gene from c. albicans has been reported. in addition to the histidine and aspartyl domains ...199910411270
the erg-3 (sterol delta14,15-reductase) gene of neurospora crassa: generation of null mutants by repeat-induced point mutation and complementation by proteins chimeric for human lamin b receptor sequences.null mutations were generated in the erg-3 gene of neurospora crassa by repeat-induced point mutation (rip). the mutants were viable, lacked ergosterol, were resistant to the steroidal glycoside alpha-tomatine and were sensitive to the phytoalexins pisatin and biochanin a. rip was frequently associated with silencing of the hph gene located adjacent to the duplicated erg-3 sequence. the silencing of hph was reversible in the two cases examined and appeared to be due to the spread of cytosine met ...199910411271
physical properties and heavy metal uptake of encapsulated escherichia coli expressing a metal binding gene (ncp).a recombinant escherichia coli expressing the neurospora crassa metallothionein gene (ncp) has previously been shown to remove low levels of cd and other metals from solution. for further development as a biosorbent, the encapsulation of the ncp is investigated by various matrices. the ncp was encapsulated in alginate, chitosan-alginate or kappa-carrageenan, and its physical properties characterized. results indicated that encapsulation in alginate resulted in fragile beads, whereas encapsulatio ...199910420333
repair of apurinic/apyrimidinic sites by uv damage endonuclease; a repair protein for uv and oxidative damage.uv damage endonuclease (uvde) initiates a novel form of excision repair by introducing a nick imme-diately 5" to uv-induced cyclobutane pyrimidine dimers or 6-4 photoproducts. here, we report that apurinic/apyrimidinic (ap) sites are also nicked by neurospora crassa and schizosaccharomyces pombe uvde. uvde introduces a nick immediately 5" to the ap site leaving a 3"-oh and a 5"-phosphate ap. apyrimidinic sites are more effectively nicked by uvde than apurinic sites. uvde also possesses 3"-repair ...199910454605
searching for the role of protein phosphatases in eukaryotic microorganisms.preference for specific protein substrates together with differential sensitivity to activators and inhibitors has allowed classification of serine/threonine protein phosphatases (pps) into four major types designated types 1, 2a, 2b and 2c (pp1, pp2a, pp2b and pp2c, respectively). comparison of sequences within their catalytic domains has indicated that pp1, pp2a and pp2b are members of the same gene family named ppp. on the other hand, the type 2c enzyme does not share sequence homology with t ...199910454741
polymorphism around cog extends into adjacent structural genes.the recombination hotspot cog overlaps a highly polymorphic 950-bp region of linkage group i in neurospora crassa. the sequence of this region in the four strains, lindegren 25a, lindegren a, emerson a and st. lawrence 74a, each differs from the others by 1.4% or more. comparison of the sequence of st. lawrence 74a and lindegren 25a each side of cog shows a high level of sequence heterology extending in both directions, including the coding sequences for his-3 and a putative gene lpl with homolo ...199910467008
the tama protein fused to a dna-binding domain can recruit area, the major nitrogen regulatory protein, to activate gene expression in aspergillus nidulans.the area gene of aspergillus nidulans encodes a gata zinc finger transcription factor that activates the expression of a large number of genes subject to nitrogen metabolite repression. the amount and activity of the area protein under different nitrogen conditions is modulated by transcriptional, post-transcriptional, and post-translational controls. one of these controls of area activity has been proposed to involve the nmra protein interacting with the dna-binding domain and the extreme c ter ...199910471703
ambient ph signal transduction in aspergillus: completion of gene characterization.completing the molecular analysis of the six pal genes of the ambient ph signal transduction pathway in aspergillus nidulans, we report the characterization of palc and palh. the derived translation product of palh contains 760 amino acids with prediction of seven transmembrane domains in its n-terminal moiety. remarkably, a palh frameshift mutant lacking just over half the palh protein, including almost all of the long hydrophilic region c-terminal to the transmembrane domains, retains some pal ...199910476033
primary structure and characterisation of a 64 kda nadh dehydrogenase from the inner membrane of neurospora crassa mitochondria.a cdna clone encoding a mitochondrial nadh dehydrogenase from neurospora crassa was sequenced. the total dna sequence encompasses 2570 base pairs and contains an open reading frame of 2019 base pairs coding for a precursor polypeptide of 673 amino acid residues. the protein is encoded by a single-copy gene located to the right side of the centromere in linkage group iv of the fungal genome. the n-terminus of the precursor protein has characteristics of a mitochondrial targeting pre-sequence. the ...199910482790
role of a white collar-1-white collar-2 complex in blue-light signal transduction.mutations in either white collar-1 (wc-1) or white collar-2 (wc-2) lead to a loss of most blue-light-induced phenomena in neurospora crassa. sequence analysis and in vitro experiments show that wc-1 and wc-2 are transcription factors regulating the expression of light-induced genes. the wc proteins form homo- and heterodimers in vitro; this interaction could represent a fundamental step in the control of their activity. we demonstrate in vivo that the wc proteins are assembled in a white collar ...199910487748
repeated ubiquitin genes in trichophyton mentagrophytes.using trichophyton mentagrophytes cdna as a template, we performed pcr amplification with the ub1s and ub1r primers for isolation of a partial fragment of t. mentagrophytes ubiquitin gene. we screened the t. mentagrophytes cdna library prepared with uni-zap phage vector by hybridization with the t. mentagrophytes ubiquitin fragment, ub1s-ub1r. the nucleotide and deduced amino acid sequences of t. mentagrophytes ubiquitin gene encoded two ubiquitin repeats in 162 bp to 624 bp. the deduced amino a ...199910489441
the 24-kda iron-sulphur subunit of complex i is required for enzyme activity.we have cloned the nuclear gene encoding the 24-kda iron-sulphur subunit of complex i from neurospora crassa. the gene was inactivated in vivo by repeat-induced point-mutations, and mutant strains lacking the 24-kda protein were isolated. mutant nuo24 appears to assemble an almost intact complex i only lacking the 24-kda subunit. however, we also found reduced levels of the nadh-binding, 51-kda subunit of the enzyme. surprisingly, the complex i from the nuo24 strain lacks nadh:ferricyanide reduc ...199910491161
complementation between mitochondrial processing peptidase (mpp) subunits from different species.mitochondrial processing peptidase (mpp), a dimer of nonidentical subunits, is the primary peptidase responsible for the removal of leader peptides from nuclearly encoded mitochondrial proteins. alignments of the alpha and beta subunits of mpp (alpha- and beta-mpp) from different species show strong protein sequence similarity in certain regions, including a highly negatively charged region as well as a domain containing a putative metal ion binding site. in this report, we describe experiments ...199910496979
the nadp-dependent glutamate dehydrogenase gene from penicillium chrysogenum and the construction of expression vectors for filamentous fungi.the gdha gene encoding the nadp-dependent glutamate dehydrogenase activity from penicillium chrysogenum has been isolated and characterized for its use in gene expression. the nucleotide sequence of a 2816-bp genomic fragment was determined, showing an open reading frame of 1600 bp interrupted by two introns, of 160 bp and 57 bp respectively, with fungal consensus splice-site junctions. the predicted amino acid sequence revealed a high degree of identity to glutamate dehydrogenase enzymes, espec ...199910499259
the nit2 nitrogen regulatory protein of neurospora: expression and stability of nit-2 mrna and protein.in neurospora crassa, nit2 is a global transcription factor that positively regulates the expression of up to 100 genes that are related to nitrogen metabolism. nit2 is responsible for the lifting of nitrogen catabolite repression when the cellular levels of glutamine or other favored nitrogen sources become limited. immunoprecipitation of the nit2 protein showed that it is constitutively expressed, although its expression level is elevated a few-fold when nitrate instead of glutamine is used as ...199910501938
characterization of a mammalian homolog of the gcn2 eukaryotic initiation factor 2alpha kinase.in eukaryotic cells, protein synthesis is regulated in response to various environmental stresses by phosphorylating the alpha subunit of the eukaryotic initiation factor 2 (eif2alpha). three different eif2alpha kinases have been identified in mammalian cells, the heme-regulated inhibitor (hri), the interferon-inducible rna-dependent kinase (pkr) and the endoplasmic reticulum-resident kinase (perk). a fourth eif2alpha kinase, termed gcn2, was previously characterized from saccharomyces cerevisia ...199910504407
circadian biology: clocks for the real world.the circadian system of neurospora crassa includes a molecular feedback loop that is entrainable by light. a recent study has shown that a second, elusive oscillator interacts with the feedback loop to drive output rhythms.199910508572
deletion of the trichodiene synthase gene of fusarium venenatum: two systems for repeated gene deletions.the trichodiene synthase (tri5) gene of fusarium venenatum was cloned from a genomic library. vectors were created in which the tri5 coding sequence was replaced with the neurospora crassa nitrate reductase (nit3) gene and with the aspergillus nidulans acetamidase (amds) gene flanked by direct repeats. the first vector was utilized to transform a nitrate reductase (niad) mutant of f. venenatum to prototrophy, and the second vector was utilized to confer acetamide utilization to the wild-type str ...199910512673
the entomopathogenic fungus metarhizium anisopliae alters ambient ph, allowing extracellular protease production and activity.ambient ph regulates the expression of virulence genes of metarhizium anisopliae, but it was unknown if m. anisopliae can regulate ambient ph. mutants of m. anisopliae altered in production of oxalic acid were evaluated for the interrelationship of ambient ph, buffering capacity added to media, growth, and generation of extracellular proteases and ammonia. wild-type and acid-overproducing mutants [acid(+)] grew almost as well at ph 8 as at ph 6, but acid-non-producing [acid(-)] mutants showed li ...199910537191
ionizing radiation-induced mutagenesis: radiation studies in neurospora predictive for results in mammalian cells.ionizing radiation was the first mutagen discovered and was used to develop the first mutagenicity assay. in the ensuing 70+ years, ionizing radiation became a fundamental tool in understanding mutagenesis and is still a subject of intensive research. frederick de serres et al. developed and used the neurospora crassa ad-3 system initially to explore the mutagenic effects of ionizing radiation. using this system, de serres et al. demonstrated the dependence of the frequency and spectra of mutati ...199910556618
comparison of the spectra of genetic damage in formaldehyde-induced ad-3 mutations between dna repair-proficient and -deficient heterokaryons of neurospora crassa.the mutagenic effects of formaldehyde (fa) have been compared in dna repair-proficient (heterokaryon 12) and dna repair-deficient (heterokaryon 59) two-component heterokaryons of neurospora crassa. the data from forward-mutation experiments were used to compare the spectra of fa-induced specific-locus mutations at two closely linked loci in the adenine-3 (ad-3) region and on the fa-induced inactivation of heterokaryotic conidia. previous studies have demonstrated that specific-locus mutations at ...199910556619
deletogenic activity of 1,2:7,8-diepoxyoctane in the salmonella typhimurium tester strain ta102.1,2:7,8-diepoxyoctane (deo), whose deletogenic activity was first demonstrated in ad-3 system of neurospora crassa and then in different species, has been tested in salmonella typhimurium tester strain ta102 (hisg428(ochre)). it was confirmed that it is a direct acting mutagen and was found that its activity is stimulated with the s9 mix. obtained his(+) revertants were screened on their response to the histidine analog, n-(2-thiazolyl)-dl-alanine (thiala). thirty-two percent of spontaneous and ...199910556620
a comparison of thermal characteristics and kinetic parameters of trehalases from a thermophilic and a mesophilic fungus.trehalases from a thermophilic fungus thermomyces lanuginosus (m(r) 145 kda) and a mesophilic fungus neurospora crassa (m(r) 437 kda) were purified to compare their thermal characteristics and kinetic constants. both trehalases were maximally active at 50 degrees c, had an acidic ph optimum and were glycoproteins (20% and 43%, w/w, carbohydrate content for t. lanuginosus and n. crassa, respectively). at their temperature optimum, their k(m) was similar (0.57 and 0.52 mm trehalose, for t. lanugin ...199910564806
a eukaryotic protein, nop-1, binds retinal to form an archaeal rhodopsin-like photochemically reactive pigment.the nop-1 gene from neurospora crassa is predicted to encode a seven-helix protein exhibiting conservation with the rhodopsins of the archaeon halobacterium salinarum. in the work presented here we have expressed this gene heterologously in the yeast pichia pastoris, obtaining a relatively high yield of 2.2 mg of nop-1 protein/l of cell culture. the expressed protein is membrane-associated and forms with all-trans retinal a visible light-absorbing pigment with a 534 nm absorption maximum and app ...199910571987
permeabilization of fungal membranes by plant defensins inhibits fungal growth.we used an assay based on the uptake of sytox green, an organic compound that fluoresces upon interaction with nucleic acids and penetrates cells with compromised plasma membranes, to investigate membrane permeabilization in fungi. membrane permeabilization induced by plant defensins in neurospora crassa was biphasic, depending on the plant defensin dose. at high defensin levels (10 to 40 microm), strong permeabilization was detected that could be strongly suppressed by cations in the medium. th ...199910584003
transfer of neurospora kalilo plasmids among species and strains by introgression.there are four different variants of the kalilo "family" of linear mitochondrial plasmids. this family is found in several heterothallic species and one pseudohomothallic species of neurospora, as well as in one homothallic species of gelasinospora. the mode of dispersal of these plasmids is not known. horizontal transmission has proved difficult to demonstrate. another possibility is transfer by introgression, and this is modelled in the present paper. we have used introgression and subsequent ...199910591968
mips: a database for genomes and protein sequences.the munich information center for protein sequences (mips-gsf), martinsried, near munich, germany, continues its longstanding tradition to develop and maintain high quality curated genome databases. in addition, efforts have been intensified to cover the wealth of complete genome sequences in a systematic, comprehensive form. bioinformatics, supporting national as well as european sequencing and functional analysis projects, has resulted in several up-to-date genome-oriented databases. this repo ...200010592176
mitop, the mitochondrial proteome database: 2000 update.mitop (http://www.mips.biochem.mpg.de/proj/medgen/mitop/) is a comprehensive database for genetic and functional information on both nuclear- and mitochondrial-encoded proteins and their genes. the five species files--saccharomyces cerevisiae, mus musculus, caenorhabditis elegans, neurospora crassa and homo sapiens--include annotated data derived from a variety of online resources and the literature. a wide spectrum of search facilities is given in the overlapping sections 'gene catalogues', 'pr ...200010592209
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
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