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quantitative transfer of the molybdenum cofactor from xanthine oxidase and from sulphite oxidase to the deficient enzyme of the nit-1 mutant of neurospora crassa to yield active nitrate reductase.an assay method is described for measurement of absolute concentrations of the molybdenum cofactor, based on complementation of the defective nitrate reductase ('apo nitrate reductase') in extracts of the nit-1 mutant of neurospora crassa. a number of alternative methods are described for preparing, anaerobically, molybdenum-cofactor-containing solutions from sulphite oxidase, xanthine oxidase and desulpho xanthine oxidase. for assay, these were mixed with an excess of extract of the nit-1 mutan ...19846234882
localization of pyruvate carboxylase in the cells of neurospora crassa.the cell wall of neurospora crassa was digested enzymatically and the cytosolic and the mitochondrial fractions were separated. the activity of pyruvate carboxylase (ec 6.4.1.1) was detected entirely in the cytosolic fraction. this indicates that the location of pyruvate carboxylase of n. crassa is in the cytosol, but is not in the mitochondria; this is different from the situation in animal tissues.19846232147
phospholipase-induced crystallization of channels in mitochondrial outer membranes.when outer membranes from neurospora crassa mitochondria are treated with low levels of phospholipase a2 under continuous dialysis, two-dimensional crystalline arrays of the pore protein component of these membranes are formed.19846322311
specific regulatory interconnection between the leucine and histidine pathways of neurospora crassa.leucine auxotrophs of neurospora fall into two discrete categories with respect to sensitivity to the herbicide, 3-amino-1,2,4-triazole. the pattern of resistance corresponds exactly to the ability to produce the leucine pathway control elements, alpha-isopropylmalate and the leu-3 product. an analysis of the regulatory response of the production of enzymes of histidine biosynthesis to alpha-isopropylmalate implicates the control elements of the leucine pathway as important components of the mec ...19846325383
large-scale isolation of the neurospora plasma membrane h+-atpase.a method for the purification of relatively large quantities of the neurospora crassa plasma membrane proton translocating atpase is described. cells of the cell wall-less sl strain of neurospora grown under o2 to increase cell yields are treated with concanavalin a to stabilize the plasma membrane and homogenized in deoxycholate, and the resulting lysate is centrifuged at 13,500g. the pellet obtained consists almost solely of concanavalin a-stabilized plasma membrane sheets greatly enriched in ...19846233916
regulation of amino acid utilization in neurospora crassa: effect of nmr-1 and ms-5 mutations.the effect of the nmr-1 and ms-5 mutations, which lead to insensitivity to glutamine-mediated nitrogen metabolite repression, was examined with respect to extracellular deaminase production by neurospora crassa. deaminase production normally requires nitrogen limitation, but these mutations eliminated this requirement and allowed production of deaminase activity under nitrogen metabolite repressing conditions. demonstration of normal glutamine transport by both strains eliminated the possibility ...19846238946
nitrogen regulation of amino acid utilization by neurospora crassa.the production of an extracellular deaminase activity involved with the utilization of amino acids as sole sources of nitrogen is under the control of the nit-2 locus of neurospora crassa. this locus is the sole major nitrogen regulatory locus described for n. crassa and is believed to encode a positive effector required for induction of activities involved with the utilization of alternate nitrogen sources. production of deaminase activity requires the lifting of nitrogen metabolite repression, ...19846238945
mitochondrial dna of the filamentous ascomycete cochliobolus heterostrophus : characterization of the mitochondrial chromosome and population genetics of a restriction enzyme polymorphism.the mitochondrial chromosome of cochliobolus heterostrophus is a circle approximately 115 kb in circumference, among the largest known from fungi. a physical map of c. heterostrophus mtdna was constructed using the restriction enzymes bamhi, ecori, and pvuli by dna-dna hybridizations with cloned or purified mtdna bamhi fragments. the following sequences were located on the mtdna map: (1) the large and small mitochondrial ribosomal rna genes (identified by heterologous hybridization to cloned neu ...198424178002
structure of the cell wall proteogalactomannan from neurospora crassa. ii. structural analysis of the polysaccharide part.the native proteoheteroglycan (phg) from mycelia of neurospora crassa contain two kinds of carbohydrate chains differing structure. the oligosaccharides containing mannose and galactofuranose are attached by o-glycosidic linkages to serine or threonine residues in the protein (j. biochem. 96, 1005-1011, 1984). the second kind of carbohydrate chain is a polysaccharide containing mannose and galactofuranose as the main sugar components. the results of structural studies with methylation and nmr an ...19846240491
chitosomes from the wall-less "slime" mutant of neurospora crassa.cell-free extracts from the wall-less slime mutant of neurospora crassa and the mycelium of wild type exhibit similar chitin synthetase properties in specific activity, zymogenicity and a preferential intracellular localization of chitosomes. the yield of chitosomal chitin synthetase from slime cells was essentially the same irrespective of cell breakage procedure (osmotic lysis or ballistic disruption)--an indication that chitosomes are not fragments of larger membranes produced by harsh (balli ...19846240239
purification of the neurospora crassa plasma membrane h+-atpase by high-pressure liquid chromatography in the presence of sodium dodecyl sulfate.current methods for purifying the mr 100,000 h+-atpase from the plasma membrane of fungi and higher plants rely on detergent solubilization followed by density gradient centrifugation. the procedure yields catalytically active enzyme of high purity but takes several days, and the yields are low. for chemical studies on the primary structure of this enzyme, an alternative more rapid procedure was sought. in this paper a method which uses a high-performance gel filtration column in the presence of ...19846240211
two intervening sequences in the atpase subunit 6 gene of neurospora crassa. a short intron (93 base-pairs) and a long intron that is stable after excision.a 3590 base-pair region of the mitochondrial genome of neurospora crassa, including the gene for atpase subunit 6 (oli2), has been sequenced. the oli2 gene is interrupted by two intervening sequences. the first intron, situated after the third codon of the gene, is 93 base-pairs long; two-thirds of this intron consist of a palindromic sequence. the second intron is 1370 base-pairs long and contains an extended open reading frame that is continuous and in frame with the upstream exon sequence. th ...19846238172
a basal unit of valine-sensitive acetolactate synthase of neurospora crassa.valine-sensitivity as well as activity of acetolactate synthase of neurospora crassa was stabilized with 1.2 m potassium phosphate buffer during extraction from mitochondria and early stages of purification, and with 20% glycerol plus 5 mm sodium pyruvate during sephadex g200 gel chromatography. the enzyme was expressed as four molecular species having the molecular weights of about 500,000, 140,000, 68,000 and 51,000, respectively. the first and the third species showed valine-sensitivity, but ...19846237648
three subunit proteins of membrane enzymes in mitochondria of neurospora crassa contain a pantothenate derivative.three proteins of the inner mitochondrial membrane of neurospora crassa were found to be covalently modified with a derivative of pantothenic acid. one of these proteins is a subunit of cytochrome c oxidase and two are subunits of the atpase-atp synthase. cells of a pantothenate auxotroph of n. crassa were labeled with [14c]pantothenic acid, and mitochondrial proteins containing radiolabeled pantothenate were detected by electrophoresis of detergent-solubilized mitochondria. mitochondria from ce ...19846088512
uv-induced recessive lethals in uvs strains of neurospora which are deficient in uv mutagenesis.the frequencies of spontaneous and uv-induced recessive lethal mutations were compared for uv-sensitive and wild-type heterokaryons of neurospora crassa. these heterokaryons were homokaryotic either for one of two alleles of uvs-3, or for uvs-6 or uvs+. for uvs-3, which is known to have mutator effects, spontaneous recessive lethals were found to be 4-6 times more frequent than observed in uvs+. after correction for clonal distribution of spontaneous mutants, an observed 2-fold increase for uvs- ...19846236366
the dna sequence and genetic organization of a neurospora mitochondrial plasmid suggest a relationship to introns and mobile elements.we have determined the complete 3581 bp sequence of the mitochondrial plasmid from neurospora crassa strain mauriceville-1c. the plasmid contains a long open reading frame that is expressed in its major transcript and could encode a hydrophilic protein of 710 amino acids. two characteristics of the plasmid--codon usage and the presence of conserved sequence elements--suggest that it is related to group i mtdna introns. the major transcripts of the plasmid are approximately full-length, colinear ...19846088081
photoregulation of some enzymes from neurospora crassa.light-grown cultures of neurospora crassa showed photoregulation of a number of enzymes. proteases and cytosolic malate dehydrogenase showed an increase in activity. there was a decrease in the activity of mitochondrial malate dehydrogenase, isocitrate dehydrogenase and cytosolic glucose-6p-dehydrogenase, isocitrate dehydrogenase and isocitrate lyase.19846239788
molecular characterization of the qa-4 gene of neurospora crassa.the qa-4 gene of neurospora crassa encodes 3-dehydroshikimate dehydratase, which catalyzes the third step of the quinic acid (qa) catabolic pathway. the enzyme has previously been purified and characterized as a monomer of approx. 37 kdal. the nucleotide sequence of the qa-4 gene is presented here and the amino acid composition and tentative nh2-terminal sequence confirm the identification of the coding region within the qa-4 dna sequence. there are no introns in the qa-4 coding region. by s1 nu ...19846241580
transcriptionally active chromatin is sensitive to neurospora crassa and s1 nucleases.we have examined the distribution of neurospora crassa and s1 nuclease cleavage products in the chromatin of the 87a7 heat shock locus of drosophila melanogaster. both of these nucleases generate single and double-strand breaks in chromatin at specific sites in the 87a7 locus. before heat induction, we find that the 5' ends of the two 87a7 hsp 70 genes contain n. crassa and s1 nuclease hypersensitive sites, while there are only a few cleavage products from elsewhere in the locus. with n. crassa ...19846096552
processing peptidase of neurospora mitochondria. two-step cleavage of imported atpase subunit 9.subunit 9 (dicyclohexylcarbodiimide binding protein, 'proteolipid') of the mitochondrial f1f0-atpase is a nuclearly coded protein in neurospora crassa. it is synthesized on free cytoplasmic ribosomes as a larger precursor with an nh2-terminal peptide extension. the peptide extension is cleaved off after transport of the protein into the mitochondria. a processing activity referred to as processing peptidase that cleaves the precursor to subunit 9 and other mitochondrial proteins is described and ...19846237909
effects of respiratory inhibitors on respiration, atp contents, and the circadian conidiation rhythm of neurospora crassa.effects of respiratory inhibitors on the circadian clock, respiratory activity, and atp content were examined in neurospora crassa. all inhibitors, potassium cyanide, sodium azide, antimycin a, and carbonyl cyanide m-chlorophenyl hydrazone (cccp), shifted the phase of the conidiation rhythm. all the phase response curves were similar and resembled that for cycloheximide, but were different from the phase response curve for light. phase shifting by azide and cccp was proportional to the lowering ...198416663893
preparation of a cell-free translation system from a wild-type strain of neurospora crassa and translation of pyruvate kinase messenger rna.a cell-free in vitro translation system exhibiting high activity has been developed from wild-type neurospora crassa mycelium. the isolation is simple and fast, and the homogenization does not appear to affect the activity of mycelial proteases and nucleases. this system is capable of supporting efficient translation of exogenously added homologous rna as demonstrated by the experiments with pk-specific mrna. in addition, it translates heterologous rna efficiently, shown by the translation of gl ...19846240997
the relationship of mo, molybdopterin, and the cyanolyzable sulfur in the mo cofactor.reconstitution of the apoprotein of the molybdoenzyme nitrate reductase in extracts of the neurospora crassa mutant nit-1 with molybdenum cofactor released by denaturation of purified molybdoenzymes is efficient in the absence of exogenous moo2-4 under defined conditions. evidence is presented that this molybdate-independent reconstitution is due to transfer of intact mo cofactor, a complex of mo and molybdopterin (mpt), the organic constituent of the cofactor. this complex can be separated from ...19846231887
acidic ribosomal proteins of neurospora crassa.neurospora crassa acidic ribosomal proteins from the high salt-ethanol extract of 80 s ribosomes have been fractionated by deae-cellulose chromatography. six acidic ribosomal proteins were purified. all resemble escherichia coli l7 and l12 in amino acid composition and molecular weight but each has a slightly different net charge at ph 3.2. four have an apparent molecular weight of approx. 14 000, and two have a molecular weight of approx. 14 800. the amino acid compositions and circular dichroi ...19846231958
activation of nit-1 nitrate reductase by w-formate dehydrogenase.formate dehydrogenase ( fdh ) from clostridium thermoaceticum is a known tungsten enzyme. fdh was tested for the presence of nitrogenase-type cofactor and nitrate reductase-type cofactor by the azotobacter vinelandii uw-45 and neurospora crassa nit-1 reconstitution assays, respectively. tungsten formate dehydrogenase (w- fdh ), containing only a small mo impurity, activated the nit-1 nitrate reductase extracts when molybdate was also added, but not when tungstate was added. these results show w- ...19846234890
accurate transcription of homologous 5s rrna and trna genes and splicing of trna in vitro by soluble extracts of neurospora.we have developed soluble extracts from neurospora crassa capable of accurately and efficiently transcribing homologous 5s rrna and trna genes. the extracts also appear to quantitatively end-process and splice the primary trna transcripts. although the extracts could not transcribe a heterologous (yeast) 5s rrna gene, they did transcribe a yeast trnaleu gene and slowly process the transcripts. in addition, we have developed a novel strategy for rapidly sequencing uniformly labelled rnas using ba ...19846235482
a leucine trna gene adjacent to the qa gene cluster of neurospora crassa.a single trnaleu gene has been localized and sequenced from neurospora crassa. it is located only 375 bp from the qa gene cluster and it is the only trna or 5s rrna gene within this cloned 37 kb region. the gene encodes a trnaleu with the anti-codon aag, and unlike the other nuclear eukaryotic trnaleu (aag) gene sequenced (from c. elegans), contains an intervening sequence of 27 bp. the neurospora trnaleu (aag) is 84% and 73% homologous respectively to the c. elegans and bovine trnaleu (aag), an ...19846235483
mutagenicity of neocarzinostatin in neurospora crassa.neocarzinostatin (ncs) is an acidic, single-chain polypeptide of 109 amino acids that has shown some antitumor activity in clinical trials. ncs is mutagenic in reca+ strains of escherichia coli, but not in reca strains; on the other hand, a defect in the nucleotide-excision-repair pathway has no effect on the mutagenicity of ncs in e. coli. similar results are seen in mammalian cells. excision-repair-deficient xeroderma pigmentosum (xp) cells repair ncs-induced dna damage at the same rate as rep ...19846236365
in vitro reconstitution of nitrate reductase activity of the neurospora crassa mutant nit-1: specific incorporation of molybdopterin.the reduced, metal-free pterin of the molybdenum cofactor has been termed molybdopterin. oxidation of any molybdopterin-containing protein in the presence or absence of iodine yields oxidized molybdopterin derivatives termed form a and form b, respectively. application of these procedures to whole cells and cell extracts has demonstrated the presence of molybdopterin in wild-type neurospora crassa, and its absence in the cofactor-deficient mutant nit-1. in order to demonstrate that the reconstit ...19846237611
a two-dimensional immunoelectrophoretic analysis of the heat-shock response exhibited by neurospora crassa cells.the heat-shock (hs) response of neurospora crassa was studied by two-dimensional (2-d) immunoelectrophoresis, in conjunction with in vivo labelling of proteins with [35s]methionine. antisera against extracts of normally grown and shocked cells were tested with both extracts as antigens. the resolution of normal cell proteins by interaction with homologous antisera yielded at least 35 immunoprecipitates. using antisera to shocked cells with normal and shocked cell extracts resolved four heat-shoc ...19846238661
structure of the cell wall proteogalactomannan from neurospora crassa. i. purification of the proteoheteroglycan and characterization of alkali-labile oligosaccharides.proteoheteroglycan (phg) was prepared from neurospora crassa cells by extraction with hot water followed by cetyltrimethylammoniumbromide fractionation. the polymer was purified by deae-cellulose chromatography followed by gel filtrations. the phg was fractionated into five subfractions containing carbohydrate (65-88%), protein (19-36%), and a trace amount of phosphate (0.3-1.9%). the sugar compositions of the fractions were similar to each other (d-mannose, 47-60%, d-galactose, 35-50%, d-glucos ...19846240490
studies on the induction of aryl hydrocarbon(benzo[a]pyrene) hydroxylase in neurospora crassa, and its suppression by sodium selenite.six fungal species were grown in the presence of benzo[a]pyrene (bp); four showed benzo[a]pyrene hydroxylase (aryl hydrocarbon hydroxylase, ahh) activity. penicillium sp. and neurospora crassa metabolized bp to a limited extent. n. crassa ahh activity was induced by bp, the major product of metabolism being 3-hydroxy-bp. both induction of ahh activity and metabolism of bp were suppressed by sodium selenite in the growth medium. two polypeptides, unique to bp-grown cells, were revealed by two-dim ...19846241764
interactions between avian myeloblastosis reverse transcriptase and trnatrp. mapping of complexed trna with chemicals and nucleases.the interactions between beef trnatrp with avian myeloblastosis reverse transcriptase have been studied by statistical chemical modifications of phosphate (ethylnitrosourea) and cytidine (dimethyl sulfate) residues, as well as by digestion of complexed trna by cobra venom nuclease and neurospora crassa endonuclease. results with nucleases and chemicals show that reverse transcriptase interacts preferentially with the d arm, the anticodon stem and the t psi stem. all these regions are located in ...19846200830
preparative-scale isolation and purification of procaryotic and eucaryotic ribosomal 5 s rna: bacillus subtilis, neurospora crassa, and wheat germ.ribosomal 5 s rna from three different organisms has been isolated in high yield and purity. without prior isolation of ribosomes, a presoak in buffer followed by phenol extraction, de-32 ion-exchange chromatography, and sephadex g-75 gel-permeation chromatography yields at least 5-10 mg of electrophoretically homogeneous 5 s rna from 100 g of cells. ribonuclease activity is eliminated by various combinations of low temperature, sodium dodecyl sulfate, phenol, and bentonite. high-molecular-weigh ...19846204554
ultrastructural aspects of cytoplasmic ribosomes from histoplasma capsulatum and blastomyces dermatitidis as revealed by heavy metal staining.cytoplasmic ribosomes were isolated and purified from sonicates of the mycelial and yeastlike growth forms of the pathogenic dimorphic fungi, histoplasma capsulatum and blastomyces dermatitidis. similar ribosomal fractions were prepared from neurospora crassa and saccharomyces cerevisiae. these latter organisms were selected as typical filamentous and yeastlike monophasic fungi, and their ribosomes were used as reference standards. high resolution electron microscopy permitted a comparison of bo ...19846206396
cytochrome b gene of neurospora crassa mitochondria. partial sequence and location of introns at sites different from those in saccharomyces cerevisiae and aspergillus nidulans.we have sequenced a 2614-base pair fragment of the neurospora crassa mitochondrial dna which contains part of the structural gene for apocytochrome b. this gene is split by at least two introns. the sequence reported here begins within one intron, extends through the next exon, another intron 1276 base pairs long, and the last exon which encodes the cooh terminus of cytochrome b. within the 254 amino acids encoded by the two exons, there is a high degree of sequence conservation, 81%, with cytoc ...19846231283
mutation tests in neurospora crassa. a report of the u.s. environmental protection agency gene-tox program.many mutation tests have been developed in neurospora crassa during the almost 40 years of its use in mutation research. these tests detect two major classes of mutation: gene mutation and meiotic nondisjunction. within the first class, forward- and reverse-mutation tests have been used. the forward-mutation tests include those that detect mutations at many loci and at specific loci. both kinds of forward-mutation tests have been done in homokaryons (n) and heterokaryons (n + n'). from the publi ...19846231482
isolation and characterization of mms-sensitive mutants of neurospora crassa.seven different mutants that show high sensitivity to mms killing were isolated and mapped at different loci. one group, mms-(sa1), mms-(sa2) and mms-(sa6), showed high sensitivity to mms but not to uv or gamma-rays. another group, mms-(sa4) and mms-(sa5), showed extremely high sensitivity to uv and mms. and mms-(sa3) and mms-(sa7) were moderately sensitive to both uv and mms. mms-(sa4) and mms-(sa1) were identified as alleles of uvs-2 and mus-7, respectively, which had been previously isolated. ...19846230534
genetically determined conidial longevity is positively correlated with superoxide dismutase, catalase, glutathione peroxidase, cytochrome c peroxidase, and ascorbate free radical reductase activities in neurospora crassa.aging of post-mitotic cells, the conidia, of neurospora crassa is defined as the time-dependent loss of viability under a constant laboratory environment which probably resembles the organism's tropical habitat; namely, at 30 degrees c, 85-100% relative humidity under white light. median lifespan is defined as the age at which survival of a conidial population has declined to 50% of that of a fully viable population at birth. a collection of short (age-) and long-lived (age+) mutants were previo ...19846319835
inhibition of superoxide dismutase by nitroprusside and electron spin resonance observations on the formation of a superoxide-mediated nitroprusside nitroxyl free radical.nitroprusside appears to inhibit the known types of superoxide dismutases irrespective of their metal prosthetic group and regardless of the source from which the enzymes were isolated. thus the copper-zinc enzyme from bovine erythrocyte or neurospora crassa behaved identically as did the manganese enzymes from escherichia coli or red alga and the iron enzyme from e. coli and a blue-green alga. the inhibition was dose dependent with a ki = 2.5 x 10(-5) for nitroprusside. nitroprusside does not b ...19846092342
analysis of the structure and transcription of the aro3 cluster gene in schizosaccharomyces pombe.by selecting activities to complement escherichia coli aro mutations, a gene responsible for the biosynthesis of aromatic amino acids in schizosaccharomyces pombe (aro3) was cloned into pbr322. three independent clones named pfna1, pfna2 and pfna3 were obtained. pfna1 could complement e. coli arod only, whereas the other two plasmids were able to complement both arod and aroe. the aro3 locus of s. pombe was found to be a gene cluster which can be subdivided into five complons, a through e (strau ...19846092844
the mitochondrial genome of the fission yeast schizosaccharomyces pombe. 2. localization of genes by interspecific hybridization in strain ade7-50h- and cloning of the genome in small fragments.a series of 18 small overlapping restriction fragments has been cloned, covering the complete mitochondrial genome of schizosaccharomyces pombe. by hybridizing mitochondrial gene probes from saccharomyces cerevisiae and neurospora crassa with restriction fragments of schizosaccharomyces pombe mitochondrial dna, the following homologous genes were localized on the mitochondrial genome of s. pombe: cob, cox1, cox2 and cox3, atpase subunit 6 and 9 genes, the large rrna gene and both types of open r ...19846094974
transformation of neurospora crassa with recombinant plasmids containing the cloned glutamate dehydrogenase (am) gene: evidence for autonomous replication of the transforming plasmid.we have characterized neurospora crassa transformants obtained with plasmid pjr2, which consists of the neurospora glutamate dehydrogenase (am) gene cloned in puc8 and an am132 host strain which contains a deletion encompassing the cloned fragment. every one of 33 transformants tested showed extreme meiotic instability: less than 1 or 2% am+ progeny were obtained in initial or successive backcrosses between am+ transformants and am132 or in intercrosses between am+ progeny. furthermore, am+ prog ...19846095037
nucleotide sequence of saccharomyces cerevisiae genes trp2 and trp3 encoding bifunctional anthranilate synthase: indole-3-glycerol phosphate synthase.saccharomyces cerevisiae anthranilate synthase:indole-3-glycerol phosphate synthase is a multifunctional hetero-oligomeric enzyme encoded by genes trp2 and trp3. trp2, encoding anthranilate synthase component i, was cloned by complementation of a yeast trp2 mutant. the nucleotide sequence of trp2 as well as that of trp3 were determined. the deduced anthranilate synthase component i primary structure from yeast exhibits only limited similarity to that of the corresponding escherichia coli subunit ...19846323449
evidence for two control genes regulating expression of the quinic acid utilization (qut) gene cluster in aspergillus nidulans.the first three steps in quinic acid degradation in aspergillus nidulans are catalysed by highly inducible enzymes encoded by a gene cluster regulated by an adjacent control region. analysis of two non-inducible mutants has been done in diploid strains, where quta8 is recessive and all three enzyme activities are fully induced in heterozygous quta8/quta+ diploids. in contrast, quta4/quta+ heterozygous diploids show semi-dominance of the mutant allele, giving markedly diminished growth on quinic ...19846374025
dna polymerases, deoxyribonucleases, and recombination during meiosis in saccharomyces cerevisiae.we utilized strains of saccharomyces cerevisiae that exhibit high efficiency of synchrony of meiosis to examine several aspects of meiosis including sporulation, recombination, dna synthesis, dna polymerase i and ii, and mg2+-dependent alkaline dnases. the kinetics of commitment to intragenic recombination and sporulation are similar. the synthesis of dna, as measured directly with diphenylamine, appears to precede the commitment to recombination. both dna polymerase i and ii activities and tota ...19846396507
isolation and manipulation of genes coding for energy-transducing enzymes from neurospora crassa and escherichia coli. 19846327431
on the origin of chromosomal aberrations in human peripheral lymphocytes in vitro. i. experiments with neurospora endonuclease and polyethylene glycol.post-treatment of mutagen-treated human peripheral lymphocytes with a single-strand specific endonuclease from neurospora crassa leads to a significant elevation of the rate of structural chromosomal aberrations. our results indicate that dna double-strand breaks (dsb) are ultimate lesions for the formation of chromosomal aberrations in the g1 and g2 phase of the cell cycle and probably also in the s-phase. post-treatment of x-irradiated g2 cells with polyethylene glycol (peg) leads to an elevat ...19846327498
the nuclear-coded subunits of yeast cytochrome c oxidase. iii. identification of homologous subunits in yeast, bovine heart, and neurospora crassa cytochrome c oxidases.sequences for the nh2-terminal halves of subunits iv, v, vi, vii, and viia from yeast cytochrome c oxidase have been determined and used to identify homologous subunits in bovine heart and neurospora crassa cytochrome c oxidases. in conjunction with the complete sequence of subunit viii (s. d. power, m. a. lochrie , t. e. patterson, and r. o. poyton (1984) j. biol. chem. 259, 6571-6574), we have been able to identify counterparts to yeast subunits iv, v, vi, and viii in bovine heart cytochrome c ...19846327686
transhyphal electrical currents in fungi.representative mycelial fungi from the phycomycete, ascomycete and basidiomycete groups (achlya bisexualis, neurospora crassa, aspergillus nidulans, schizophyllum commune and coprinus cinereus) all generated steady electrical currents around their hyphal tips; the generation of a transhyphal ion current may therefore be a universal characteristic of hyphal growth. as with all other tip growing organisms, positive current always entered apically and left distally; non-growing hyphae did not drive ...19846520604
molybdopterin in carbon monoxide oxidase from carboxydotrophic bacteria.the carbon monoxide oxidases (coxs) purified from the carboxydotrophic bacteria pseudomonas carboxydohydrogena and pseudomonas carboxydoflava were found to be molybdenum hydroxylases, identical in cofactor composition and spectral properties to the recently characterized enzyme from pseudomonas carboxydovorans (o. meyer, j. biol. chem. 257:1333-1341, 1982). all three enzymes exhibited a cofactor composition of two flavin adenine dinucleotides, two molybdenums, eight irons and eight labile sulfid ...19846582059
a mitochondrial reading frame which may code for a second form of atpase subunit 9 in aspergillus nidulans.the nucleotide sequence of a 74 codon reading frame from the aspergillus nidulans mitochondrial genome is presented. the derived amino acid sequence displays typical features of dicyclohexylcarbodiimide (dccd) binding proteins and is 84% homologous with a mitochondrial reading frame that potentially encodes an atpase subunit 9 polypeptide in neurospora crassa. however, in a. nidulans, as in n. crassa, there is strong biochemical and genetic evidence that this subunit is in fact nuclearly-encoded ...198424177948
cloning and preliminary characterization of a molybdenum cofactor gene of neurospora crassa.a neurospora crassa library, constructed in a derivative of the plasmid pbr322 (prk9), was used to transform two e. coli ch1d molybdenum cofactor mutants (ch1d, ch1d::mu). subsequently, one transformant from each of three independent transformation experiments was restriction mapped. all three transformants had an identical n. crassa dna insert (4.2 kb). southern blot analysis with one of the plasmids (pmoco, 1:4) showed hybridization to a single band of n. crassa genomic dna. when pmoco plasmid ...198424177998
biochemical characterization of the molybdenum cofactor mutants of neurospora crassa: in vivo and in vitro reconstitution of nadph-nitrate reductase activity.molybdenum cofactor (moco) mutants of neurospora crassa lack both nadph-nitrate reductase and xanthine dehydrogenase activity. in vivo and in vitro studies to further characterize these mutants are now reported. the moco mutants nit-9a and nit-9b are capable of growing, albeit poorly, with nitrate as the sole nitrogen source, provided high levels of molybdate are present. the moco mutants nit-9a, nit-9b and nit-9c, but not nit-1, nit-7 or nit-8, have significant levels of nadph-nitrate reductase ...198424177997
structure and function of the trp3 gene of saccharomyces cerevisiae: analysis of transcription, promoter sequence, and sequence coding for a glutamine amidotransferase.the structure and function of the trp3 gene of saccharomyces cerevisiae were analyzed. subcloning of an original 4.8 kb bamhi dna fragment, carrying the yeast trp3 gene, allowed for a localization of the gene on a 2.5 kb clai/bamhi fragment. transcription was found to proceed from the clai site towards the bamhi site. three major transcription start sites were determined at positions -92, -87, and -81 by s1-mapping. the synthesis of the trp3 gene is regulated by the general control, and was foun ...198424177735
biogenesis of cytochrome c in neurospora crassa. synthesis of apocytochrome c, transfer to mitochondria and conversion to holocytochrome c.1. precipitating antibodies specific for apocytochrome c and holocytochrome c, respectively, were employed to study synthesis and intracellular transport of cytochrome c in neurospora in vitro. 2. apocytochrome c as well as holocytochrome c were found to be synthesized in a cell-free homogenate. a precursor product relationship between the two components is suggested by kinetic experiments. 3. apocytochrome c synthesized in vitro was found in the post-ribosomal fraction and not in the mitochondr ...1984215405
voltage-dependent channels found in the membrane fraction of corn mitochondria.transmembrane channels have been found in the membrane fraction of corn (zea mays w64an) mitochondria that exhibit a remarkable resemblance to the voltage dependent anion-selective channels (vdac) located in the outer membrane of animal (rattus norvegicus), protist (paramecium aurelia), and fungal (neurospora crassa) mitochondria. the channels in corn were demonstrated to be essentially identical to vdac channels in three characteristic properties: (a) single channel conductance magnitude, (b) w ...198516664537
genetic analysis of transformation in a microconidiating strain of neurospora crassa.we have characterized neurospora crassa transformants obtained with plasmid pdv1001 bearing the cloned catabolic dehydroquinase (qa-2+) gene (hughes et al. 1983a) and fluffy 268 host strain producing only uninucleate microconidia allowing to isolate individual transformation products. the percentage of transformed nuclei in the mycelium and their stability were determined by genetic analysis of microconidia produced on selective or non-selective medium. about half of the transformants originatin ...19852970312
identification of molybdoproteins in clostridium pasteurianum.cells of clostridium pasteurianum whose n source is switched from nh3 to n2 accumulate large amounts of molybdenum beginning 1.5 h before the detection of nitrogenase activity. anaerobic multiphasic gel electrophoresis and anion-exchange chromatography were used to identify the molybdoproteins and molybdenum-containing components present in n2-fixing cells. in addition to molybdate, six distinct 99mo-labeled species were detected, i.e., a membrane fragment, the mofe protein of nitrogenase, forma ...19853857223
involvement of a particular species of beta-tubulin (beta 3) in conidial development in aspergillus nidulans.strains of aspergillus containing the bena22 mutation are resistant to benomyl for vegetative growth but do not produce conidia. to test whether conidiation involved an additional benomyl-sensitive tubulin (i.e., was mediated by a tubulin other than the tubulins coded for by the bena locus), a collection of mutants was produced that formed conidia in the presence of benomyl, i.e., were conidiation-resistant (cr-) mutants. we analyzed the tubulins of these cr- mutants using two-dimensional gel el ...19853897246
identification and functional analysis of beta-tubulin genes by site specific integrative transformation in aspergillus nidulans.we have cloned two different beta-tubulin sequences from the filamentous fungus aspergillus nidulans. each was used in the construction of transforming plasmids that carry the pyr4 gene of neurospora crassa. we used these plasmids to transform a pyrg-strain of aspergillus to uridine prototrophy. both plasmids were shown to integrate site specifically into the homologous chromosomal sequences. we then used transformant strains in genetic crosses to demonstrate that one of the cloned beta-tubulin ...19853897247
cloning and characterization of the three enzyme structural genes qutb, qutc and qute from the quinic acid utilization gene cluster in aspergillus nidulans.heterologous dna probes from the quinic acid gene cluster (qa) in neurospora crassa (schweizer 1981) have been used to isolate the corresponding gene cluster (qut) from aspergillus nidulans cloned in a phage lambda vector. n. crassa probes for each of the three enzyme structural genes in the cluster have been used to identify the corresponding genes within the a. nidulans cloned dna. the three genes are in the same relative sequence [dehydrogenase (1), qa-3 = qutb; dehydratase (3), qa-4 = qutc; ...19853916726
specific inhibition of mitochondrial protein synthesis influences the amount of complex i in mitochondria of rat liver and neurospora crassa directly.specific inhibition of mitochondrial protein synthesis reduces the oxidation rate of nadh-linked substrates in rat liver as well as in neurospora crassa mitochondria. the present study shows that this is due to the fact that inhibition of mitochondrial protein synthesis leads to a decrease of the concentration of active complex i. therefore, these results demonstrate that at least one of the genes for the subunits of complex i is localized on mitochondrial dna.19853934002
the mitochondrial genome of the fission yeast schizosaccharomyces pombe. the cytochrome b gene has an intron closely related to the first two introns in the saccharomyces cerevisiae cox1 gene.the dna sequence of the cob region of the schizosaccharomyces pombe mitochondrial dna has been determined. the cytochrome b structural gene is interrupted by an intron of 2526 base-pairs, which has an open reading frame of 2421 base-pairs in phase with the upstream exon. the position of the intron differs from those found in the cob genes of saccharomyces cerevisiae, aspergillus nidulans or neurospora crassa. the sch. pombe cob intron has the potential of assuming an rna secondary structure almo ...19854046021
sequence of the n-terminal formic acid fragment and location of the n-ethylmaleimide-binding site of the phosphate transport protein from beef heart mitochondria.the n-terminal formic acid fragment (fa1) of the n-[3h]ethylmaleimide-labeled and carboxymethylated bovine mitochondrial phosphate transport protein (ptpn*cm) has been purified and completely sequenced: nh2-ala-val-glu-glu-gln-tyr-ser-cys-asp-tyr10-gly-ser-gly-arg-phe- phe-ile-leu-cys- gly20-leu-gly-gly-ile-ile-ser-cys-gly-thr-thr30-his-thr -ala-leu-val-pro-leu-asp- -leu-val40-lys-cys(n-[3h]ethylmaleimide)-arg-met-gln-val-asp- cooh. by thermolysin digestion of fa1 and high-performance liquid chr ...19854066697
primary structure of tyrosinase from streptomyces glaucescens.the complete amino acid sequence of streptomyces glaucescens tyrosinase is reported. the molecule consists of 273 amino acids and has a mr of 30 900 including two copper atoms. the primary structure was determined by a combination of amino acid and dna sequence analysis. peptide sequence information was derived from the cyanogen bromide, tryptic, and thermolytic fragments of apotyrosinase by automated edman degradation and aminopeptidase m and carboxypeptidase c digestions. the nucleotide sequen ...19853002431
the mitochondrial urf1 gene in neurospora crassa has an intron that contains a novel type of urf.in neurospora crassa, a 2670 base-pair segment of the mitochondrial dna was sequenced including a gene homologous to the mammalian urf1 that was recently shown to encode a subunit of the respiratory chain nadh dehydrogenase complex. urf1 of n. crassa is interrupted by an intron of 1118 base-pairs that divides the protein-coding sequence into two exons of 636 and 480 base-pairs length, respectively. the deduced urf1 polypeptide of 371 residues was aligned with that of other eukaryotes, revealing ...19853003362
characterization of neurospora crassa cyclic amp phosphodiesterase activated by calmodulin.activation of cyclic amp phosphodiesterase i by brain or neurospora calmodulin was studied. the stimulation required micromolar concentrations of ca2+, and it was observed at cyclic amp concentrations between 0.1 and 500 microm. activation was blocked by edta and some neuroleptic drugs such as chlorpromazine and fluphenazine. these drugs inhibit the elongation of n. crassa wild-type aerial hyphae. these results reinforce the evidence towards the recognition of ca2+-calmodulin as one of the syste ...19853004404
cloning and characterization of the multifunctional his-3 gene of neurospora crassa.we have cloned the his-3 gene of neurospora crassa and determined its nucleotide sequence. the gene specifies a protein of 863 amino acids (aa) and contains a 59-bp intron which interrupts aa 800, a proline residue. the 5' end of the his-3 transcript is heterogeneous with major starts 122 and 124 bp upstream from the start codon. there are three possible polyadenylation sites, 119, 120 and 121 bp after the uaa stop codon. the protein shows two regions of homology to the yeast his4 gene which cor ...19853005109
nucleotide sequence of the mitochondrial cytochrome oxidase subunit ii gene in the yeast hansenula saturnus.the gene for subunit ii of cytochrome oxidase in the yeast hansenula saturnus was previously shown to be located on a 1.7 kb hindiii-bamhi fragment of mitochondrial dna (lawson and deters, accompanying paper). in this paper, we report the nucleotide sequence of a large part of this fragment, covering the coding region of the subunit ii gene, designated coxii, and its 5' and 3' flanking regions. the coding region of the coxii gene consists of a continuous open reading frame, 744 nucleotides long, ...19852836090
stimulation of beta(1----3)glucan synthetase of various fungi by nucleoside triphosphates: generalized regulatory mechanism for cell wall biosynthesis.particulate fractions from the taxonomically diverse fungi achlya ambisexualis, hansenula anomala, neurospora crassa, cryptococcus laurentii, schizophyllum commune, and wangiella dermatitidis were found to catalyze the time-dependent incorporation of glucose from udp-[14c]glucose into a water-insoluble material. the reaction was stimulated by bovine serum albumin. the product was characterized as beta(1----3)glucan on the basis of its resistance to alpha- and beta-amylase and susceptibility to b ...19853156122
protein changes during the asexual cycle of neurospora crassa.a method for synchronizing conidiation and isolating large numbers of cells at discrete stages of conidia development is described. using two-dimensional gel electrophoresis, we analyzed the protein profiles of mycelia, aerial hyphae, and conidia and observed that the concentration of 14 polypeptides increase and 38 decrease during the asexual cycle. twelve polypeptides were present in extracts of aerial hyphae or conidia, but not mycelia, suggesting that they may be conidiation specific. the pr ...19853157863
primary structure of the trpc gene from aspergillus nidulans.we have determined the structure and complete nucleotide sequence of the trifunctional trpc gene from the ascomycetous fungus aspergillus nidulans. results from rna gel blot analyses showed that this gene encodes two size classes of polyribosomal, poly (a)+rnas with approximate lengths of 2,400 and 2,600 nucleotides. s1 nuclease protection studies demonstrated that the distribution into the two size classes is due to selection of alternative sites for polyadenylation. the transcription units con ...19853158796
effect of the homokaryotic or heterokaryotic state of the uvs-2 allele in neurospora crassa on mitomycin c-induced killing and ad-3 mutation.mitomycin c (mc) was tested for its killing and mutagenic activities in the ad-3 forward-mutation test in neurospora crassa. the test was conducted in 4 dikaryons of n. crassa in order to determine the effect of the uvs-2 allele, which causes a defect in nucleotide excision repair, on mc-induced killing and ad-3 mutation. these dikaryons were homokaryotic for uvs-2+ (h-12), homokaryotic for uvs-2 (h-59), and heterokaryotic for uvs-2/uvs-2+ (h-70 and h-71). mc induced killing and ad-3 mutation in ...19853158811
ornithine transcarbamylase from neurospora crassa: purification and properties.ornithine transcarbamylase catalyzes the synthesis of citrulline from carbamyl phosphate and ornithine. this enzyme is involved in the biosynthesis of arginine in many organisms and participates in the urea cycle of mammals. the biosynthetic ornithine transcarbamylase has been purified from the filamentous fungus, neurospora crassa. it was found to be a homotrimer with an apparent subunit molecular weight of 37,000 and a native molecular weight of about 110,000. its catalytic activity has a ph o ...19853159341
primary structure of copper-zinc superoxide dismutase from neurospora crassa.the complete amino acid sequence of copper-zinc superoxide dismutase from neurospora crassa is reported. the subunit consists of 153 amino acids and has a mr of 15,850. the primary structure was determined by automated and manual sequence analysis of peptides obtained by digestions of the carboxymethylated and aminoethylated enzyme with trypsin and thermolysin. the protein is devoid of tryptophan and methionine and displays a free amino terminus. comparison of the amino acid sequence with those ...19853160699
functional effects and cross-reactivity of antibody to purified subunit b (uncf protein) of escherichia coli proton-atpase.subunit b (uncf protein) of the proton-atpase (f1f0) of escherichia coli was purified from membranes of strain an1460 (unc+). antibody to purified subunit b was raised in rabbits. it reacted with f1-depleted membranes and blocked f1 binding. bound antibody had no effect on proton transport through f0. f1-depleted membranes competed with purified subunit b for antibody in an enzyme-linked immunosorbent assay. f1-depleted membranes which had been pretreated with trypsin or preincubated with satura ...19852857549
size of the plasma membrane h+-atpase from neurospora crassa determined by radiation inactivation and comparison with the sarcoplasmic reticulum ca2+-atpase from skeletal muscle.using radiation inactivation, we have measured the size of the h+-atpase in neurospora crassa plasma membranes. membranes were exposed to either high energy electrons from a van de graaff generator or to gamma irradiation from 60co. both forms of radiation caused an exponential loss of atpase activity in parallel with the physical destruction of the mr = 104,000 polypeptide of which this enzyme is composed. by applying target theory, the size of the h+-atpase in situ was found to be approximatel ...19852862141
plasmid recovery from transformants and the isolation of chromosomal dna segments improving plasmid replication in neurospora crassa.the efficient recovery of plasmid dna from neurospora crassa transformants is described. lithium acetate-treated spores were transformed with plasmid dna and grown in mass in liquid culture. the resulting mycelial growth was harvested and plasmid dna was extracted and used to transform e. coli to ampicillin resistance. although at low frequency, routine recovery of plasmid psd3 which carries the neurospora qa-2+ gene and pbr322 sequences has been demonstrated. about 10% of the recovered plasmids ...19852967124
nucleotide sequence of the gdh gene coding for the nadp-specific glutamate dehydrogenase of saccharomyces cerevisiae.the isolation of the saccharomyces cerevisiae gene for nadp-dependent glutamate dehydrogenase (nadp-gdh) by cross hybridization to the neurospora crassa am gene, known to encode for nadp-gdh is described. two dna fragments selected from a yeast genomic library in phage lambda gt11 were shown by restriction analysis to share 2.5 kb of common sequence. a yeast shuttle vector (cv13) carrying either to the cloned fragments complements the gdh- strain of s. cerevisiae and directs substantial overprod ...19852932370
carbamoyl-phosphate synthetases from neurospora crassa. immunological relatedness of the enzymes from neurospora, bacteria, yeast, and mammals.neurospora crassa contains two carbamoyl-phosphate synthetases: a mitochondrial enzyme (cps-a) which supplies carbamoyl phosphate for arginine biosynthesis, and a nuclear enzyme whose product is used for the synthesis of pyrimidines. we have prepared antiserum against a highly purified preparation of the large subunit of cps-a and have used the antiserum to demonstrate that the large subunit is, like most mitochondrially localized proteins, initially synthesized as a higher molecular weight prec ...19852932445
isolation and characterization of the aspergillus niger trpc gene.the aspergillus niger trpc gene was isolated by complementation experiments with an escherichia coli trpc mutant. plasmid dna containing the a. niger trpc gene transforms an aspergillus nidulans mutant strain, defective in all three enzymatic activities of the trpc gene, to trp+, indicating the presence of a complete and functional trpc gene. southern blot analysis of dna from these trp+ transformants showed that plasmid dna was present but that this dna was not integrated at the site of the chr ...19852936650
influence of neurospora endonuclease on trenimon-induced structural chromosomal aberrations and sister-chromatid exchanges in human peripheral lymphocytes and chinese hamster ovary cells.human peripheral lymphocytes and chinese hamster ovary cells were treated in the g1 phase of the cell cycle with the trifunctional alkylating agent trenimon (trn) and post-treated with a single-strand specific endonuclease from neurospora crassa (ne). trn induces chromosomal aberrations of the chromatid type (ca) and sister-chromatid exchanges (sce). ne post-treatment leads to an elevation of the frequencies of ca but not of sces. this indicates that trn induced ca are the result of dna double-s ...19852985980
cloning and characterization of the rdna repeat unit of podospora anserina.dna coding for ribosomal rna in podospora anserina has been cloned and was found as a tandemly repeated 8.3 kb sequence. the cloned rdna was characterized by restriction endonuclease mapping. the location of 5.8s, 18s and 28s rrna coding regions was established by dna-rna hybridization and s1 nuclease mapping. the organization of p. anserina rrna genes is similar to that of neurospora crassa and aspergillus nidulans. the rdna unit does not contain the sequence coding for 5s rna.19852987647
an equilibrium between distorted and undistorted dna in the adult chicken beta a-globin gene.we have used single strand specific nucleases to map dna distortion in the adult chicken beta a-globin gene. we have detected two structures of that kind and have mapped nuclease-cutting sites at one base resolution. one prominent site is centered at -190 relative to the rna capping site and is positioned at the center of a stretch of contiguous c residues. the second site is near the first intron/exon junction (+620) and appears as a series of discrete 1-base-long enzyme-cutting sites. based up ...19852989280
nucleotide sequence of yeast gdh1 encoding nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase.the yeast gdh1 gene encodes nadp-dependent glutamate dehydrogenase. this gene was isolated by complementation of an escherichia coli glutamate auxotroph. nadp-dependent glutamate dehydrogenase was overproduced 6-10-fold in saccharomyces cerevisiae bearing gdh1 on a multicopy plasmid. the nucleotide sequence of the 1362-base pair coding region and 5' and 3' flanking sequences were determined. transcription start sites were located by s1 nuclease mapping. regulation of gdh1 was not maintained when ...19852989290
structure of a neurospora rna polymerase i promoter defined by transcription in vitro with homologous extracts.a neurospora in vitro transcription system has been developed which specifically and efficiently initiates transcription of a cloned neurospora crassa ribosomal rna gene by rna polymerase i. the initiation site of transcription (both in vitro and in vivo) appears to be located about 850 bp from the 5' end of mature 17s rrna. however, the primary rrna transcripts are normally cleaved very rapidly at a site 120-125 nt from the 5' end in vitro and in vivo. the nucleotide sequence surrounding the in ...19852989792
cloning and expression of the fbc operon encoding the fes protein, cytochrome b and cytochrome c1 from the rhodopseudomonas sphaeroides b/c1 complex.the gene for the fes protein of the rhodopseudomonas sphaeroides b/c1 complex was identified by means of cross-hybridization with a segment of the gene encoding the corresponding fes protein of neurospora crassa. plasmids (prsf1-14) containing the cross-hybridizing region, covering in total 13.5 kb of chromosomal dna, were expressed in vitro in a homologous system. one rsf plasmid directed the synthesis of all three main polypeptides of the r. sphaeroides b/c1 complex: the fes protein, cytochrom ...19852990901
comparison of the mitochondrial endonucleases from neurospora crassa and saccharomyces cerevisiae.the endonucleases from neurospora crassa and saccharomyces cerevisiae are not closely related antigenically. they also differ with respect to their activity at ph 8, their degree of hydrophobicity, and their sensitivity to elevated temperatures. however, the two nucleases have similar specific activities, are inhibited by edta, and have nearly identical substrate specificities. since the enzymes also have the same mode of action and intracellular location, these similarities may indicate that th ...19852992732
cloning of the aro cluster gene of neurospora crassa and its expression in escherichia coli.we have constructed a phage, lambda ncl, which comprises a 4.0 kb hindiii insert of neurospora dna into the immunity region of the vector lambda 598. lambda ncl complements the arod6 mutation of e. coli, permitting the formation of galaxy plaques on medium lacking aromatic supplements, and transforms an aro-9 qa-2 neurospora mutant to prototrophy at a low frequency. low levels of 5-dehydroquinate hydrolyase (e.c.4.2.1.10.), with properties unlike those of the catabolic isoenzyme that is coded by ...19852993794
isolation and sequence analysis of a cdna encoding the atp/adp translocator of zea mays l.a cdna complementary to the mrna for the atp/adp translocator of maize (zea mays l.) has been identified by virtue of hybridisation with the homologous gene from yeast. the cloned cdna has been shown by dna sequence analysis to contain an open reading frame of 954bp., which encodes a polypeptide of molecular weight 40,519. this polypeptide exhibits a high degree of homology to the translocator polypeptides of beef heart and neurospora crassa mitochondria.19852994015
excision-amplification of mitochondrial dna during senescence in podospora anserina. dna sequence analysis of three unique "plasmids".during senescence in the filamentous fungus podospora anserina, specific regions of the mitochondrial genome, termed sendna are excised, ligated and amplified. we have cloned in their entirety three such autonomously replicating plasmids, alpha, beta and epsilon sendna. none of these plasmids displayed cross-hybridization nor did we detect any significant dna homology by computer analysis. the complete dna sequence of the 2.5 kb alpha, the 5.5 kb epsilon and about 3.4 kb of the 9.8 kb beta sendn ...19852997455
calmodulin and ca2+-dependent cyclic amp phosphodiesterase activity in trypanosoma cruzi.calmodulin has been purified from trypanosoma cruzi epimastigote forms by ion-exchange chromatography, gel filtration and affinity chromatography on 2-chloro-10-(3-aminopropyl)phenotiazine-sepharose. upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the factor showed a polypeptide band with an apparent molecular weight of 16 000. in addition, cyclic amp phosphodiesterase activity from t. cruzi epimastigote forms was purified by ion-exchange chromatography and affinity chromatograph ...19852999589
correct removal by splicing of a neurospora intron in yeast.processing of intron-containing nuclear messenger rnas in yeast require an internal conserved sequence (ics) element, uacuaac. similar elements (ugcuagac) have been identified in sequences interrupting nuclear genes of the related ascomycete neurospora crassa. to examine the structural splicing requirements in yeast, we constructed hybrid genes containing the intron of the neurospora histone h3 gene and cloned them into high copy number yeast vectors. subsequently we analyzed the rnas transcribe ...19852999703
development of a high-frequency transforming vector for aspergillus nidulans.the pyr4 gene of neurospora crassa, which codes for orotidine-5'-phosphate decarboxylase, is capable of transforming an aspergillus nidulans pyrg mutant by chromosomal integration, despite low homology between the transforming dna and the recipient genome. integration of pfb6, a plasmid carrying pyr4 and capable of replication in escherichia coli, was not observed at the pyrg locus. the efficiency of transformation was considerably enhanced (50-100 fold) by inclusion in the transforming vector o ...19853000883
heterogeneity of 5s rna in fungal ribosomes.neurospora crassa has at least seven types of 5s rna genes (alpha, beta, gamma, epsilon, delta, zeta, and eta) with different coding regions. a high resolution gel electrophoresis system was developed to separate minor 5s rna's from the major 5s rna (alpha). a study of several neurospora crassa strains, four other species in the genus neurospora, members of two closely related genera, and three distantly related genera demonstrated that 5s rna heterogeneity is common among fungi. in addition, di ...19852579431
probing fungal mitochondrial evolution with trna.sequence data are now available for almost the entire complement of mitochondrial rrnas from five fungi: schizosaccharomyces pombe, saccharomyces cerevisiae, toropulis glabrata, aspergillus nidulans and neurospora crassa. analysis of these data show that the five mitochondria can be related to a common ancestor. the unusually high similarity between some s. pombe mt trnas may be due to a process similar to gene conversion. using the number of differences between trna pairs as a measure of the ev ...19852417640
changes in gene expression elicited by amino acid limitation in neurospora crassa strains having normal or mutant cross-pathway amino acid control.the effects of amino acid limitation on gene expression have been investigated in neurospora crassa strains carrying normal (cpc-1+) or mutant (cpc-1) alleles at a locus implicated in cross-pathway amino acid control. electrophoresis and fluorography were used to reveal the patterns of label incorporation into polypeptides in vivo, or after in vitro translation of extracted mrnas. in a cpc-1+ strain at least 20% of detectable in vitro translation products showed relative increases in incorporati ...19852412092
growth and macromolecular synthesis phenotypes of a heat-sensitive mutant strain, rip-1, of neurospora crassa.a heat-sensitive mutant of neurospora crassa, strain 4m(t), was isolated using ultraviolet-light mutagenesis followed by the inositol-less death enrichment technique. the heat-sensitivity is the result of a single gene mutation which maps to the distal end of the right arm of linkage group ii. the mutation defines the rip-1 gene locus. both conidial germination and mycelial extension are inhibited in the mutant at 35 degrees c and above (the nonpermissive temperature) but prolonged incubation at ...19852412091
isolation and structural organization of the neurospora crassa copper metallothionein gene.the neurospora crassa copper metallothionein gene was cloned and its complete nucleotide sequence is reported. enriched metallothionein mrna was used as a template for cdna synthesis, primed by a metallothionein-specific, synthetic undecanucleotide. the sequence of the cdna obtained allowed the synthesis of a unique 21-mer which was used to screen a genomic dna library of n. crassa. in agreement with the published amino acid sequence, the gene codes for a polypeptide 26 amino acid residues in le ...19852932331
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