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active transport of l-aspartic acid in neurospora crassa. 19714258794
an inducible amino acid transport system in neurospora crassa. 20144258482
effect of l-sorbose on polysaccharide synthetases of neurospora crassa (glycogen- -1,3-glucan-morphology-cell wall-digitonin-particulate enzymes).neurospora glycogen synthetase (ec 2.4.1.11) occurs in 100,000 x g particles. the two forms (glucose-6-phosphate dependent-independent) of glycogen synthetase were solubilized and separated by digitonin treatment of the 100,000 x g particles. glucan synthetase activity of neurospora was found only in a cell-wall preparation. these two enzymes have been characterized in relation to the paramorphogenic action of sorbose. sorbose-grown cultures showed a marked decrease in the specific activity of b ...19724258315
genetic control of recombination in neurospora crassa: correlated regulation in unlinked chromosome intervals. 19714258281
some details and effects of the premeiotic controls of recombination frequencies in neurospora crassa. 19714258173
disruption of an amino acid transport mutant of neurospora crassa by kcl.a double amino acid transport-deficient mutant (pm (-)nb) of neurospora crassa is shown to be altered in the molecular structure of its cell wall or membrane. this alteration was revealed by a high degree of cellular disruption and cell-cell interaction following extraction by a high molar concentration of kcl.19724258064
kinetic properties of phosphofructokinase of neurospora crassa. 19724258060
novel mutation causing derepression of several enzymes of sulfur metabolism in neurospora crassa.a group of enzymes of sulfur metabolism (arylsulfatase, cholinesulfatase, and a number of others) are normally repressed in neurospora crassa by an abundant supply of a "favored" sulfur source such as methionine or inorganic sulfate. a mutant called scon(c) was isolated in which the formation of each of these enzymes is largely or completely nonrepressible. the structural genes for three of these enzymes have been mapped; scon(c) is not linked to any of them. it is also not linked to cys-3, anot ...19724257980
long-term cryogenic storage of neurospora crassa spores. 19714257765
neurospora crassa ribosomes: effects of bound peptidyl-trna on dissociation into subunits and on phenylalanine polymerization at low mg 2+ concentration. 19714257627
cysteine toxicity in neurospora crassa: the mechanism of counteraction by amino acids. 19714256680
magnesium and polyamine levels in neurospora crassa mycelia. 19714256640
cysteine toxicity in neurospora crassa: comparison of counteraction by sulphur amino acids and iron. 19704252565
a succinic dehydrogenase activity in "mesosomes" of neurospora crassa. 19714250787
inhibition of protein synthesis in neurospora crassa by 8-azaguanine. 19694245416
the in vivo iron donor for haem synthesis in neurospora crassa. 19684239950
phosphoribosyl-aminoimidazole-succinocarboxamide synthetase from neurospora crassa. i. partial purification and properties. 19694239091
[mechanism of action of thiopicolinic acid anilide in neurospora crassa]. 19684237100
laminarinase of neurospora crassa. i. enzyme activity associated with conidia & conidial wall. 19684234673
[griseocarnin, an antifungal antibiotic isolated from streptoverticillium criseocarneum (ia-7527)].we have been studying the antagonistic activity of the streptoverticillium griseocarneum, strain ia-7527. it produces a strong antifungal substance which we named griseocarnin. its antimicrobial power to neurospora crassa, rhodotorula, rubra, hansenula suaveolens and candida albicans deserves to be exceled. the isolated substance was compared with other antifungal antibiotics presenting close similarity with the heptaens ascosin and pa-150. the production, extraction and purification methods as ...19744220132
carcinogenic and noncarcinogenic polycyclic hydrocarbons in neurospora crassa and chinese hamster cells: their photodynamic effects. 19704193459
biochemical and histochemical localization of invertase in neurospora crassa during conidial germination and hyphal growth.the intracellular localization of neurospora invertase, an enzyme partially secreted and partially retained by neurospora at the cell periphery, was investigated. a cell wall fraction was isolated, to which 24% of the cell-bound invertase was firmly attached. a sensitive osmiophilic stain for invertase was developed and used in conjunction with the technique of indirect immunofluorescence to follow the pattern of invertase localization during the development of neurospora from the germination of ...19704192564
isolation and characterization of chromatin from neurospora crassa.different preparations of chromatin isolated from mycelia of neurospora crassa were analyzed for dna-associated rna and proteins. the uv absorption spectra, the ultrastructure of chromatin, and the amino acid composition of the acid-extractable proteins were studied. the protein:dna ratios range from 1.5 to 2.8; the rna:dna ratios range from 0.5 to 1.24. uv absorption shows a macimum at 259 mmicro and a minimum at 238-239 mmicro. the e280/e260 ranges from 0.59 to 0.70. electron microscopy reveal ...19694186412
nuclear division in neurospora crassa during conidiation and germination. 19674168108
slow conformational changes of a neurospora glutamate dehydrogenase studied by protein fluorescence.1. the nadp-dependent glutamate dehydrogenase of neurospora crassa undergoes slow reversible structural transitions, with half-times in the order of a few minutes, between active and inactive states. the inactive state of the enzyme, which predominates at ph values below 7.0, has an intrinsic tryptophan fluorescence 25% lower than that of the active state, which predominates at ph values above 7.6. the inactive state can be activated either by an increase in ph or by addition of activators such ...19744156826
chorismate synthase of neurospora crassa: a flavoprotein. 19744155270
amino-acid sequence of nadp-specific glutamate dehydrogenase of neurospora crassa.a tentative primary structure of the nadp-specific glutamate dehydrogenase [l-glutamate: nadp oxidoreductase (deaminating), ec 1.4.1.4] from neurospora crassa has been determined. the proposed sequence contains 452 amino-acid residues in each of the identical subunits of the hexameric enzyme. comparison of the sequence with that of the bovine liver enzyme reveals considerable homology in the amino-terminal portion of the chain, including the vicinity of the reactive lysine, with only shorter str ...19744155068
purification and properties of the neurospora crassa assimilatory nitrite reductase. 19744154942
role of the redox state of nicotinamide adenine dinucleotides in the in vivo regulation of inositol biosynthesis in neurospora crassa. 19734147849
properties of chorismate synthase in neurospora crassa. 19734146266
circadian rhythms in neurospora: spatial differences in pyridine nucleotide levels.a growing colony of a mutant strain of neurospora crassa had two morphologically distinct areas which were formed as a result of a rhythmic spore-forming (conidiation) process. the total pyridine nucleotide content of these two areas was the same, but the levels of nadh, nadph, and nadp were lower in the conidiating area, while the nad level was higher. these biochemical differences in the adjacent areas of a single colony were only found in newly formed areas, and were not a permanent record. i ...19734144759
the inhibition of the neurospora crassa nitrate reductase complex by metal-binding agents. 19734144237
comparison of the respiratory chain of neurospora crassa wild type and the mi-mutants mi-1 and mi-3. 19734144164
[studies on the conidial differentiation of "neurospora crassa". v. ultrastructure of the macroconidiogenous sequence (author's transl)]. 19734138206
some observations of ascospores of neurospora crassa made with a scanning electron microscope.scanning electron micrographs of ascospores of neurospora crassa reveal two of the structures which develop during germination and outgrowth: (i) a germination pore and (ii) the probable site of initiation of hyphal cell wall synthesis.19724115466
carbamoyl phosphate compartmentation in neurospora: histochemical localization of aspartate and ornithine transcarbamoylases.carbamoyl phosphate is required for arginine and pyrimidine synthesis. in the arginine pathway, it is used in the ornithine transcarbamoylase (ec 2.1.2.1) reaction; in the pyrimidine pathway, it is used in the aspartate transcarbamoylase (ec 2.1.3.2) reaction. in neurospora crassa, two pathway-specific enzymes catalyze the synthesis of carbamoyl phosphate, and two path-specific pools of carbamoyl phosphate are maintained. histochemical studies show that ornithine transcarbamoylase is located in ...19724114857
virus-like particles in certain slow-growing strains of neurospora crassa. 19724110285
end-product regulation of the tryptophan-nicotinic acid pathway in neurospora crassa.the regulation of the tryptophan-nicotinic acid pathway in neurospora crassa was examined with mutants (nic-2, nic-3) which require nicotinamide for growth. the accumulation of n-acetylkynurenin and 3-hydroxyanthranilic acid by these mutants served to estimate the level of function of the early reactions in the pathway. in still cultures, maximal accumulation occurred with media containing growth-limiting amounts of nicotinamide; the accumulation of intermediates was almost negligible with nicot ...20144107105
demonstration of dihydro-orotate dehydrogenase in neurospora crassa hyphae with a cytochemical procedure. 19714106595
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
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
isolation and partial characterization of two antifungal proteins from barley.we have developed a simple assay for detecting antifungal compounds utilizing impregnated paper discs on agar to inhibit mycelial spread of an indicator organism, trichoderma reesei. using this assay we have isolated and purified to apparent homogeneity two antifungal proteins from dehusked barley grain. both proteins are present at high concentrations: over 10 mg of each protein can be isolated per 100 g of grain. the first protein has a molecular weight of 30 000 and is identical to the 30 kda ...19863942788
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
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
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
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 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
purification and properties of nitrate reductase from mitsuokella multiacidus.nitrate reductase of mitsuokella multiacidus (formerly bacteroides multiacidus) was solublized from the membrane fraction with 1% sodium deoxycholate and purified 40-fold by immunoaffinity chromatography on the antibody-affi-gel 10 column. the preparation showed a major band (86% of total protein) with enzyme activity and a minor band on polyacrylamide gel after disc electrophoresis in the presence of 0.1% triton x-100. sodium dodecyl sulfate-polyacrylamide gel electrophoresis gave a major band, ...19863711052
molybdopterin cofactor from methanobacterium formicicum formate dehydrogenase.the molybdopterin cofactor from the formate dehydrogenase of methanobacterium formicicum was studied. the cofactor was released by guanidine denaturation of homogeneous enzyme, which also released greater than 80% of the molybdenum present in the enzyme. the anoxically isolated cofactor was nonfluorescent, but after exposure to air it fluoresced with spectra similar to those of described molybdopterin cofactors. aerobic release from acid-denatured formate dehydrogenase in the presence of i2 and ...19863700335
extraction and purification of molybdenum cofactor from milk xanthine oxidase.molybdenum cofactor (mocofactor) is extracted efficiently, free of impurities and in high concentrations, by acid treatment of xanthine oxidase and subsequent incubation of the precipitate with phosphate buffer containing edta, molybdate and oxygen. it is suggested that cofactor is bound to the enzyme via hydrophobic forces as well as via an oxygen-sensitive mechanism. upon extraction, the capability to complement the apo nitrate reductase of neurospora crassa nit-1 can be conserved only in the ...19873691496
a long polypyrimidine/polypurine tract induces an altered dna conformation on the 3' coding region of the adjacent myosin heavy chain gene.a long (147 base pairs), natural a.t rich polypyrimidine/polypurine tract has been found 55 base pairs downstream of a chicken embryonic myosin heavy chain (mhc) gene. analysis at the nucleotide level of nicks induced by s1 and neurospora crassa nucleases indicate that this long interrupted polypyrimidine/polypurine tract exists in an alternate dna structure in vitro at ph 4.5 and ph 7.5 in both supercoiled and linear plasmid dna. the polypyrimidine/polypurine tract induces this alternate struct ...19873671071
complete amino-acid sequence of a functional unit from a molluscan hemocyanin (helix pomatia).from the beta c-hemocyanin (beta c-hc) of the vineyard snail, helix pomatia, the functional unit d (mr approximately equal to 50,000-55,000) was isolated by limited proteolysis and gel chromatography. a small quantity of functional unit d was obtained intact, but the major part in the form of two peptides (mr approximately equal to 43,000 and 10,000, respectively) connected by a disulfide bridge. after reduction and carboxymethylation, these were separated from each other and cleaved by conventi ...19873620107
a protein required for splicing group i introns in neurospora mitochondria is mitochondrial tyrosyl-trna synthetase or a derivative thereof.the nuclear cyt-18 mutants of neurospora crassa are defective in splicing a number of group i introns in mitochondria. here, cloning and sequencing of the cyt-18 gene show that it contains an open reading frame having significant homology to bacterial tyrosyl-trna synthetases. biochemical and genetic experiments lead to the conclusions that the cyt-18 gene encodes mitochondrial tyrosyl-trna synthetase, that mutations in this gene inhibit splicing directly, and that mitochondrial tyrosyl-trna syn ...19873607872
biosynthesis of thiamin. different biosynthetic routes of the thiazole moiety of thiamin in aerobic organisms and anaerobic organisms.the nitrogen atom of glycine was incorporated into the thiazole moiety of thiamin in the aerobic microorganisms bacillus subtilis, pseudomonas putida, saccharomyces cerevisiae, mucor racemosus, neurospora crassa, and emericella nidulans. it was not incorporated in the case of the facultative anaerobic microorganisms escherichia coli and enterobacter aerogenes, which, however, did incorporate the nitrogen atom of tyrosine. these results show that aerobic microorganisms and facultative anaerobic m ...19873566774
further characterization of trimethylamine n-oxide reductase from escherichia coli, a molybdoprotein.escherichia coli trimethylamine n-oxide (tmao) reductase i, the major enzyme among inducible tmao reductases, was purified to homogeneity by an improved method including heat treatment, ammonium sulfate precipitation, and chromatographies on bio-gel a-1.5m, deae-cellulose, and reactive blue-agarose. the molecular weight was estimated by gel filtration to be approximately 200,000. a single subunit peptide with a molecular weight of 95,000 was found by sodium dodecyl sulfate-polyacrylamide gel ele ...19863528139
activation in vitro of respiratory nitrate reductase of escherichia coli k12 grown in the presence of tungstate. involvement of molybdenum cofactor.the chlorate-resistant (chlr) mutants are pleiotropically defective in molybdoenzyme activity. the inactive derivative of the molybdoenzyme, respiratory nitrate reductase, present in the cell-free extract of a chlb mutant, can be activated by the addition of protein fa, the probable active product of the chlb locus. protein fa addition, however, cannot bring about the activation if 10 mm sodium tungstate is included in the culture medium for the chlb strain. the inclusion of a heat-treated prepa ...19863525161
molybdenum cofactor: a compound in the in vitro activation of both nitrate reductase and trimethylamine-n-oxide reductase activities in escherichia coli k12.nitrate reductase (nitrite: (acceptor) oxidoreductase, ec 1.7.99.4) and trimethylamine n-oxide reductase (nadh : trimethylamine-n-oxide oxidoreductase, ec 1.6.6.9) activities were reconstituted by incubation of the association factor fa (the putative product of the chlb gene) with the soluble extract of the chlb mutant grown anaerobically in the presence of trimethylamine n-oxide. when soluble extracts of the chlb mutant grown on 10 mm sodium tungstate, a molybdenum competitor, were used in comp ...19863524687
transformation of aspergillus oryzae using the a. niger pyrg gene.a transformation system for aspergillus oryzae based on the orotidine-5'-phosphate decarboxylase gene (pyrg) was developed. transformation frequencies of up to 16 transformants per microgram of dna were obtained with the vector pab4-1, which carries the pyrg gene of a. niger. southern blotting analysis showed that vector dna sequences were integrated into the chromosomal dna, in various copy numbers and presumably at different sites. efficient cotransformation of an unselectable gene was also sh ...19873481025
development of a homologous transformation system for aspergillus niger based on the pyrg gene.the development of a homologous transformation system for aspergillus niger is described. the system is based on the use of an orotidine-5'-phosphate decarboxylase deficient mutant (pyrg) and a vector, pab4-1, which contains the functional a. niger pyrg gene as a selection marker. transformation of the a. niger pyrg mutant with pab4-1 resulted in the appearance of stable pyr+ transformants at a frequency of 40 transformants per microgram of dna. in 90% of these transformants integration had occu ...19873472035
sequence organization and putative regulatory elements in the 5s rrna genes of two higher plants (vigna radiata and matthiola incana).the tandemly arranged and clustered highly repeated 5s rrna genes are investigated for two plants belonging to different higher plant families: matthiola incana (brassicaceae, dilleniidae, rosidae; 3600 5s rrna genes/n) shows a homogeneous repeat size of 510 bp, whereas vigna radiata (mung bean, former phaseolus aureus, fabaceae, rosidae; approx. 4300 5s rrna genes) has a repeat size of 215 bp. the mung-bean 5s rrna coding region starts 5' with agg and ends with cct; matthiola starts with ggg an ...19883371663
neurospora mitochondria contain an acyl-carrier protein.mitochondria of neurospora crassa were found to contain a protein which was labelled with [14c]pantothenic acid and which carried an acyl group. this protein, when purified 6000-fold, closely resembled the bacterial and chloroplast acyl-carrier protein(s) [acp(s)] in its physical and chemical properties. the predominant acyl group esterified to the purified protein was 3-hydroxytetradecanoate, as determined by gas chromatographic mass spectrometry. the amino acid sequence of the tryptic peptide ...19883360014
behaviour of recombinant plasmids in aspergillus nidulans: structure and stability.a pyrg- aspergillus strain was transformed with plasmid pdjb-1, derived from pbr325 by insertion of the neurospora crassa pyr4 gene (orotidine 5'-phosphate carboxylase), giving mitotically unstable transformants. aspergillus dna which acted as an "autonomously replicating sequence" (ars) in yeast was inserted into pdjb-1 and the resulting construct, pdjb12.1, gave mitotically stable transformants when introduced into aspergillus. transformants obtained with pdjb-1 and pdjb12.1 gave few pyr- prog ...19863329034
cloning, mapping and molecular analysis of the pyrg (orotidine-5'-phosphate decarboxylase) gene of aspergillus nidulans.we have modified the transformation procedures of ballance et al. [biochem. biophys. res. commun. 112 (1983) 284-289] to give increased rates of transformation in aspergillus nidulans. with the modified procedures we have been able to complement pyrg89, a mutation in the orotidine-5'-phosphate decarboxylase gene of a. nidulans, by transformation with a library of wild-type (wt) sequences in pbr329. we have recovered, by marker rescue from one such transformant, a plasmid (pjr15) that carries an ...19873328733
mitochondrial binding of a protein affected in mutants resistant to the microtubule inhibitor podophyllotoxin.specific antibodies to a protein p1 mr approximately equal to 63,000) from chinese hamster ovary cells, which is affected in mutants resistant to the microtubule inhibitor, podophyllotoxin, and behaves like a microtubule-related protein by certain criteria [14], have been raised. the antibody reacts specifically with the p1 protein in one- and two-dimensional immunoblots, and a cross-reacting protein of similar molecular mass and electrophoretic mobility is also found in cells from various verte ...19873319627
purification and properties of the major nuclease from mitochondria of saccharomyces cerevisiae.the vast majority of nuclease activity in yeast mitochondria is due to a single polypeptide with an apparent molecular weight of 38,000. the enzyme is located in the mitochondrial inner membrane and requires non-ionic detergents for solubilization and activity. a combination of heparin-agarose and cibacron blue-agarose chromatography was employed to purify the nuclease to approximately 90% homogeneity. the purified enzyme shows multiple activities: 1) rnase activity on single-stranded, but not d ...19883286639
purification and properties of escherichia coli dimethyl sulfoxide reductase, an iron-sulfur molybdoenzyme with broad substrate specificity.dimethyl sulfoxide reductase, a terminal electron transfer enzyme, was purified from anaerobically grown escherichia coli harboring a plasmid which codes for dimethyl sulfoxide reductase. the enzyme was purified to greater than 90% homogeneity from cell envelopes by a three-step purification procedure involving extraction with the detergent triton x-100, chromatofocusing, and deae ion-exchange chromatography. the purified enzyme was composed of three subunits with molecular weights of 82,600, 23 ...19883280546
sequencing and overexpression of the escherichia coli aroe gene encoding shikimate dehydrogenase.the escherichia coli aroe gene encoding shikimate dehydrogenase was sequenced. the deduced amino acid sequence was confirmed by n-terminal amino acid sequencing and amino acid analysis of the overproduced protein. the complete polypeptide chain has 272 amino acid residues and has a calculated mr of 29,380. e. coli shikimate dehydrogenase is homologous to the shikimate dehydrogenase domain of the fungal arom multifunctional enzymes and to the catabolic quinate dehydrogenase of neurospora crassa.19883277621
microorganisms associated with mouldiness of dried yam chips and their prevention.the broad objective of this study was to isolate and identify the microorganisms causing mouldiness of stored yam chips and to look for ways of preventing the problem. microorganisms isolated included aspergillus flavus, a. glaucus, a. nidulans, a. niger, a. ochraceous, a. tamarii, a. candidus, penicillium oxalicum, trichoderma longibrachyatum, rhizopus nigricans, cylindrocarpon radicicola, neurospora crassa, botryodiplodia theobromae, bacillus subtilis, bacillus cereus, erwinia carotovora and s ...19883231261
molybdenum cofactor deficiency in a patient previously characterized as deficient in sulfite oxidase.the metabolic status of a patient previously characterized as deficient in sulfite oxidase was reexamined applying new methodology which has been developed to distinguish between a defect specific to the sulfite oxidase protein and sulfite oxidase deficiency which arises as a result of molybdenum cofactor deficiency. urothione, the metabolic degradation product of the molybdenum cofactor, was undetectable in urine samples from the patient. analysis of molybdenum cofactor levels in fibroblasts by ...19883219233
the gene for rat uncoupling protein: complete sequence, structure of primary transcript and evolutionary relationship between exons.the complete nucleotide sequence of rat uncoupling protein gene has been determined. 4.5 kb of the 5'-flanking region have also been sequenced. the site of transcription start as well as 3'-end extremities were identified. transcription unit spans 8.4 kb and contains 6 exons and 5 introns. uncoupling protein as well as related mitochondrial carriers such as adp/atp carrier and phosphate carrier has a triplicated structure and each repeat of uncoupling protein corresponds to 2 exons. two gene dup ...19883202878
dna sequence and organization of the mitochondrial nd1 gene from podospora anserina: analysis of alternate splice sites.earlier, we reported that the nd1 mitochondrial gene of podospora anserina is mosaic, containing at least three class i introns. we have now completed the sequence of the nd1 gene and have determined that it contains four class i introns of 1,820, 2,631, 2,256 and 2,597 bp with the entire gene complex containing 10,505 bp, only 1,101 of which are exon sequences. introns 1 and 3 appear to be related in that their open reading frames (orfs) exhibit extensive amino acid sequence similarity and like ...19883197134
structural gene for ornithine decarboxylase in neurospora crassa.to define the structural gene for ornithine decarboxylase (odc) in neurospora crassa, we sought mutants with kinetically altered enzyme. four mutants, pe4, pe7, pe69, and pe85, were isolated. they were able to grow slowly at 25 degrees c on minimal medium but required putrescine or spermidine supplementation for growth at 35 degrees c. the mutants did not complement with one another or with odc-less spe-1 mutants isolated in earlier studies. in all of the mutants isolated to date, the mutations ...19853162095
effect of phosphate levels on the synthesis of acid phosphatases (ec 3.1.3.2) in neurospora crassa. 19853161781
gene disruption by transformation in neurospora crassa.to establish conditions which might permit deliberate gene disruptions in neurospora crassa, we studied transformation with linear dna fragments. the transformation frequency observed was increased about twofold in comparison with that obtained with circular plasmid dna. however, only a low proportion, approximately 10%, of the integration events occurred at the homologous site, whereas most integrations of transforming dna took place in nonhomologous regions. it was also found that multiple int ...19853160929
carotenoid synthesis in neurospora crassa. 19853160913
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
identification of the polypeptide encoded by the urf-1 gene of neurospora crassa mtdna.two peptides, potentially representing antigenic determinants of a proposed gene product, were synthesized. the peptide sequences were deduced from the nucleotide sequence of the unidentified reading frame (urf)1 of the neurospora crassa mitochondrial genome. specific antisera to the synthetic peptides were produced. the antibodies recognized a single polypeptide species with an apparent relative molecular mass of about 30 000. the mitochondrial origin of this polypeptide was verified by in vivo ...19853160590
rna processing in neurospora crassa mitochondria: transfer rnas punctuate a large precursor transcript.the pattern of transcripts arising from a large contiguous portion of the neurospora crassa mitochondrial genome and the processing of these transcripts have been investigated. evidence is presented for the transcription of a single, at least 12.5 kb, precursor rna that contains sequences corresponding to the apocytochrome b (cob), trnacys, cytochrome oxidase subunit 1 (co i), trnaarg and unidentified reading frame (urf) 1 genes. the two trna sequences serve as punctuation signals for the cleava ...19853160579
the kinetics of changes in the fatty acid composition of neurospora crassa lipids after a temperature increase.the fatty acid composition of the total lipids, phospholipids and neutral lipids of log-phase shaker cultures of the bd (band) strain of neurospora crassa, were measured every 2 h for an 8-h period following a temperature increase from 22 to 40 degrees c. for purposes of comparison, the fatty acid composition was also measured when cultures were grown from inoculation at temperatures between 22 and 40 degrees c. in the phospholipids, the temperature jump produced, over a 4-6 hour span, a linear ...20113159434
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
distinct roles of putrescine and spermidine in the regulation of ornithine decarboxylase in neurospora crassa.we wished to identify metabolic signals governing changes in ornithine decarboxylase (l-ornithine carboxy-lyase, ec 4.1.1.17) activity in neurospora crassa. by manipulations of the ornithine supply and by the use of inhibitors of the polyamine pathway, we found that spermidine negatively governs formation of active ornithine decarboxylase and that putrescine promotes inactivation of the enzyme. direct addition of putrescine or spermidine to cycloheximide-treated cells confirmed the role of putre ...19853159019
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
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
heat shock response of neurospora crassa: protein synthesis and induced thermotolerance.at elevated temperatures, germinating conidiospores of neurospora crassa discontinue synthesis of most proteins and initiate synthesis of three dominant heat shock proteins of 98,000, 83,000, and 67,000 mr and one minor heat shock protein of 30,000 mr. postemergent spores produce, in addition to these, a fourth major heat shock protein of 38,000 mr and a minor heat shock protein of 34,000 mr. the three heat shock proteins of lower molecular weight are associated with mitochondria. this exclusive ...19853158641
mutagenicity of 2-aminopurine, 6-n-hydroxylaminopurine, and 2-amino-n6-hydroxyadenine in neurospora crassa.these data from our experiments with 3 purine analogs reveal striking differences in mutagenic potency. it seems highly likely that these analogs substitute readily for adenine and that they cause mutations in the main part, and in the case of aha perhaps predominantly, by mispairing with cytosine. the most potent mutagens are those with the hydroxylamino group at the c6 position (aha and hap). of these, the most potent is the analog with an amino group in the c2 position (aha). the most interes ...19853158304
isolation and characterization of genes differentially expressed during conidiation of neurospora crassa.a neurospora crassa genomic dna library was screened with a cdna probe enriched in sequences expressed in conidiating cultures. clones were isolated that preferentially hybridized to this probe versus a second cdna probe complementary to polyadenylated rna isolated from mycelia. twelve clones contained unique sequences that hybridized to 22 transcripts, 19 of which accumulated preferentially in conidiating cultures. eight transcripts were present in higher levels in conidiating cultures than in ...19853157864
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
the 3-dehydroquinate synthase activity of the pentafunctional arom enzyme complex of neurospora crassa is zn2+-dependent.we have demonstrated the co-purification in constant ratio of all five activities of the pentafunctional arom enzyme complex from neurospora crassa. progressive inactivation of the 3-dehydroquinate synthase component of the purified enzyme complex by chelating agents was blocked by the substrate, 3-deoxy-d-arabino-heptulosonate 7-phosphate, but not by the cofactor nad+. full activity was restored at zn2+ concentrations as low as 0.05 nm. atomic absorption data indicated that the intact enzyme co ...19853157372
identification and chromosomal distribution of 5s rrna genes in neurospora crassa.the 5s rrna genes of neurospora crassa, unlike those of most organisms, are not tandemly arranged, and they are found imbedded in a variety of unique sequences. the 5s rrna regions of most of the genes are of one type, alpha; however, several other "isotypes" (beta, gamma, delta, zeta, and eta) are also found. we asked whether neurospora 5s rrna genes are dispersed on a chromosomal scale and whether genes of different isotypes are spatially segregated. we identified, by dna sequencing, 5s rrna g ...19853157192
unsaturated fatty acid isomers: effects on the circadian rhythm of a fatty-acid-deficient neurospora crassa mutant.the fatty acids oleic, linoleic, and linolenic, each of which has a cis double bond at the delta 9 position, are known to lengthen the circadian period of conidiation (spore formation) of strains of neurospora crassa carrying the cel mutation. cel confers a partial fatty acid requirement on the organism and has been used to promote incorporation of exogenous fatty acids. to test whether a physical effect imparted by the cis double bonds, such as increased membrane fluidity, is critical for the p ...19853156558
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
inducible beta-oxidation pathway in neurospora crassa.an inducible beta-oxidation system was demonstrated in a particulate fraction from neurospora crassa. the activities of all individual beta-oxidation enzymes were enhanced in cells after a shift from a sucrose to an acetate medium. the induction was even more pronounced in transfer to a medium containing oleate as sole carbon and energy source. since an acyl-coenzyme a (coa) dehydrogenase was detected instead of acyl-coa oxidase, the former enzyme seems to catalyze the first step of the beta-oxi ...19853155714
circadian rhythms in neurospora crassa: interactions between clock mutations.mutations at four loci in neurospora crassa that alter the period of the circadian rhythm have been used to construct a series of double mutant strains in order to detect interactions between these mutations. strains carrying mutations at three of these loci have altered periods on minimal media: prd-1, several alleles at the olir (oligomycin resistance) locus and four alleles at the frq locus. a mutation at the fourth locus, cel, which results in a defect in fatty acid synthesis, also leads to ...19853155701
a pantothenate derivative is covalently bound to mitochondrial proteins in neurospora crassa.the presence of protein-bound pantothenate in neurospora crassa was investigated by labelling a pantothenate auxotroph (pan-2) with [14c]pantothenate and examining mycelial homogenates on dodecyl sulfate/polyacrylamide gels. five peaks of radioactivity were found, with apparent molecular masses of 200, 140, 22, 19, and 9 kda. the 200-kda peak was identified as fatty acid synthetase, based on its absence in a fatty acid synthetase mutant. the 22-kda and 19-kda peaks co-purified with mitochondrial ...19853155682
evolutionary relationships among copper proteins containing coupled binuclear copper sites.the amino acid sequences of different copper proteins containing coupled binuclear copper centers are compared. hemocyanins from arthropods and molluscs and tyrosinases from three different species were found to share a highly homologous region in the c-terminal parts. this region contains three invariant histidines previously identified as ligands to cu(b) in panulirus interruptus hemocyanin by x-ray crystallography (gaykema et al., nature 309, 23-29 (1984]. in contrast, the ligand environment ...19883136463
the simple repeat poly(dt-dg).poly(dc-da) common to eukaryotes is absent from eubacteria and archaebacteria and rare in protozoans.genomic dna from a wide variety of prokaryotic and eukaryotic organisms has been assayed for the simple repeat sequence poly(dt-dg).poly(dc-da) by southern blotting and dna slot blot hybridizations. consistent with findings of others, we have found the simple alternating sequence to be present in multiple copies in all organisms in the animal kingdom (e.g., mammals, reptiles, amphibians, fish, crustaceans, insects, jellyfish, nematodes). the tg element was also found in lower eukaryotes (sacchar ...19863127653
structural studies of the vacuolar membrane atpase from neurospora crassa and comparison with the tonoplast membrane atpase from zea mays.the h+-translocating atpase located on vacuolar membranes of neurospora crassa was partially purified by solubilization in two detergents, triton x-100 and n-hexadecyl-n,n-dimethyl-3-ammonio-1-propanesulfonate, followed by centrifugation on sucrose density gradients. two polypeptides of mr approximately equal to 70,000 and approximately equal to 62,000 consistently migrated with activity, along with several minor bands of lower molecular weight. radioactively labeled inhibitors of atpase activit ...19863079903
mitotic gene conversion, reciprocal recombination and gene replacement at the bena, beta-tubulin, locus of aspergillus nidulans.we have developed a procedure for determining the rates of mitotic recombination of an interrupted duplication created by integration of transforming plasmid sequences at the bena, beta-tubulin, locus of aspergillus nidulans. transformation of a strain carrying a benomyl-resistant bena allele with plasmid aipgm4, which carries the wild-type bena allele and the pyr4 (orotidine-5'-phosphate decarboxylase) gene of neurospora crassa, creates an interrupted duplication with plasmid sequences flanked ...19883054484
molecular cloning, identification and transcriptional analysis of genes involved in acetate utilization in neurospora crassa.four neurospora crassa genomic clones have been selected as hybridizing much more strongly to labelled mrna isolated from acetate-grown mycelium than to mrna from sucrose-grown mycelium. hybridization of restriction fragments with acetate-specific mrna or cdna has been used to delimit the transcribed region(s) of each clone. the transcription of all four clones is strongly induced by transfer of growing mycelium from sucrose to acetate as sole carbon source. in wild-type mycelium, mrnas correspo ...19883054423
promoter recognition by escherichia coli rna polymerase. influence of dna structure in the spacer separating the -10 and -35 regions.escherichia coli rna polymerase contacts promoter dna at two regions (the -10 and -35 regions) which are separated by a segment of spacer dna. previously we showed that base substitutions in the spacer dna can affect promoter strength both in vitro and in vivo; these results were interpreted to reflect altered structural properties of the substituted dnas. here we provide experimental support for this interpretation. the pattern of cleavage of the promoters with neurospora crassa endonuclease an ...19883050126
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