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subunit 1 of cytochrome oxidase from neurospora crassa: nucleotide sequence of the coding gene and partial amino acid sequence of the protein.a partial protein sequence (223 residues) of cytochrome oxidase subunit 1 from neurospora crassa has been established. the nucleotide sequence of a cloned mitochondrial dna segment, including the structural gene coding for the mature subunit 1 (co i locus) was determined. in contrast to the situation in yeast, the co i locus in n. crassa is not interrupted by long intervening sequences. a polypeptide of 555 residues with a mol. wt. of 61 000 has been deduced from the reading frame established by ...19826327266
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
mutagenicity screening with fungal systems.several fungal species have been used for mutagenicity screening: aspergillus nidulans, saccharomyces cerevisiae, and neurospora crassa. the eukaryotic nature of these organisms with typical chromosomes in a nucleus and their mitotic and meiotic mode of nuclear division have been the basis for the development of test systems that cover the full spectrum of genetic changes typical for eukaryotes. it is possible to detect simple point mutations and also grosser structural chromosomal alterations. ...19836349475
identification of an iron uptake system specific for coprogen and rhodotorulic acid in escherichia coli k12.with the lac operon fusion technique, mutants were isolated in two genes that specify two outer membrane proteins designated fhue (76 k) and fiu (83 k). the synthesis of both proteins was increased under low iron growth conditions. the fhue-protein was shown to be necessary for iron uptake via coprogen, an iron chelator produced by certain fungi, e.g. neurospora crassa. in addition to fhuee the genes fhucdb, tonb and exbb were necessary for iron coprogen uptake. the gene fhue was mapped between ...19836353165
distribution of a corticosteroid-binding protein in candida and other fungal genera.using [3h]corticosterone as a probe, corticosteroid-binding protein (cbp) was detected in eight out of eight isolates of candida albicans, of both a and b serotypes. the apparent dissociation constant (kd) in the various isolates ranged between 8 and 19 nm; the binding capacity varied from 122 to over 2400 fmol (mg cytosol protein)-1. there was no correlation between the amount or affinity of cbp and isolate virulence for murine hosts. further analysis revealed demonstrable cbp in six out of six ...19836355389
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
genetic toxicology of ethylenediaminetetraacetic acid (edta).edta and its salts have a number of applications in medicine and pharmacy. edta is used to remove calcium from the human body, and serves as an anticoagulant and as a detoxicant after poisoning by heavy metals. it is often used in analytical chemistry for complexometric titrations and many other purposes. because the compound is of rather low toxicity, it is used as a food additive to bind metal ions. edta affects the inhibition of dna synthesis in primary cultures of mammalian cells. this may b ...19836406880
n,n'-dicyclohexylcarbodiimide binds specifically to a single glutamyl residue of the proteolipid subunit of the mitochondrial adenosinetriphosphatases from neurospora crassa and saccharomyces cerevisiae.the n,n'-dicyclohexylcarbodiimide-binding proteolipid subunit of the mitochondrial adenosinetriphosphatases (atp phosphohydrolase, ec 3.6.1.3) of neurospora crassa and saccharomyces cerevisiae were purified from mitochondria incubated with the radioactively labeled inhibitor. the specifically labeled subunit was cleaved with cyanogen bromide and n-bromosuccinimide, and the resultant fragments were separated by gel chromatography in the presence of 80% (vol/vol) formic acid. the n,n'-dicyclohexyl ...19806444724
the hexokinases from wild-type and morphological mutant strains of neurospora crassa. 19806444874
3-amino-1,2,4-triazole is an inhibitor of protein synthesis on mitoribosomes in neurospora crassa.the effects of the herbicide, 3-amino-1,2,4-triazole, an inhibitor of heme synthesis in rat liver, have been examined in the mold neurospora crassa. the drug is a potent inhibitor of the growth of the mold and produces biochemical changes identical to those produced by chloramphenicol. 3-amino-1,2,4-triazole, like chloramphenicol, is a direct and specific inhibitor of protein synthesis on mitoribosomes. a decrease in the levels of mitochondrial proteins which are completely or partly made on mit ...19806445208
effects of ph on the mutagenicity of sodium azide in neurospora crassa and salmonella typhimurium.sodium azide at various ph values did not cause a significant increase in the frequency of forward mutation above the control frequency at the adenine-3 (ad-3) region in resting conidia and in conidia from growing cultures of heterokaryons 12 and 59 of neurospora crassa. conidia from ad-3 mutants were plated with sodium azide at various ph values, and no obvious increase in reverse mutation above the controls was observed. data are presented showing that sodium azide at ph 3 is inactivating coni ...19806445502
mutagenic activity of tetryl, a nitroaromatic explosive, in three microbial test systems.the military explosive n-methyl-n,2,4,6-tetranitroaniline (tetryl) is a direct acting mutagen in three microbial mutagenicity test systems at concentrations as low as 5 microgram/ml. in two of these systems (neurospora crassa and salmonella typhimurium) it caused base-pair substitution reversions. in the third (saccharomyces cerevisiae d4) it caused mitotic gene conversions at two loci. three nonaromatic explosives (petn, rdx, hmx) did not exhibit mutagenic activity in the ames salmonella test. ...19806445609
the extraction of inositol-containing phospholipids and phosphatidylcholine from saccharomyces cerevisiae and neurospora crassa.by use of fungi grown in the presence of [3h]-inositol and [14c]choline, we have explored methods for the quantitative extraction of inositol-containing phospholipids and phosphatidylcholine. slightly alkaline mixtures of both ethanol-water and ethanol-diethylether-water at elevated temperatures were shown to effectively extract these lipids from intact saccharomyces cerevisiae and neurospora crassa. some previously published procedures fail to completely extract to very polar phosphoinositol-co ...19806445928
[influence of cultivation temperature on the development of functionally active photoregulatory systems in cells of neurospora crassa and physarum polycephalum].illumination with a visible light is known to induce accumulation of colored carotenoid pigments in mycelial cells on neurospora crassa and sporulation of starved macroplasmodia of physarum polycephalum. in both microorganisms, the temperature, at which they are cultivated in the dark, controls manifestation of the photoregulation processes in the cells subsequently exposed to light. the maximal rate of photoinduced accumulation of carotenoids in n. crassa is observed, when the cells are grown i ...19806446087
mutagenesis at the ad-3a and ad-3b loci in haploid uv-sensitive strains of neurospora crassa. i. development of isogenic strains and spontaneous mutability.7 different mutations that confer sensitivity to inactivation by ultraviolet light have been investigated for their effect on spontaneous mutation at the ad-3a and ad-3b loci in haploid strains of neurospora crassa. the collection and development of strains isogenic to wild-type 74a is described as well as experiments to determine the effects of each mutation on the spontaneous ad-3 mutation frequency. 6 of the strains showed spontaneous ad-3 mutant frequencies not significantly different from t ...19806446681
mutagenesis at the ad-3a and ad-3b loci in haploid uv-sensitive strains of neurospora crassa. ii. comparison of dose-response curves for inactivation and mutation induced by uv.uv-induced inactivation and induction of mutations at the ad-3a and ad-3b loci of neurospora crassa have been compared among 7 different uv-sensitive strains and a standard wild-type strain. the 7 strains show varying degrees of sensitivity to uv-induced inactivation, with the relative sensitivity being: uvs-2 greater than uvs-3 greater than uvs-4 greater than uvs-6 greater than upr-1 greater uvs-5 greater than uvs-1. studies on the induction of ad-3 mutants by uv show that the 2 excision-repair ...19806446682
protein synthesis by hybrid ribosomes reconstructed from rabbit reticulocyte ribosomal core-particles and amphibian or fungal split-proteins.it was shown that high-salt (2.75 m-nh4cl/69mm-mgcl2) shock treatment at 0 degrees c of the larger subparticles (l-subparticles) of rabbit, xenopus laevis and neurospora crassa cytoplasmic ribosomes yielded split-protein fractions that were not only functionally equivalent but also interchangeable. thus, although the remaining core-particles were inactive in both the puromycin reaction and in poly(u)-directed polyphenylalanine synthesis, activity was restored when they were combined with either ...19806446904
secondary structure predictions for the nad-specific glutamate dehydrogenase of neurospora crassa.from the amino acid sequences of the three known fragments of the nad-specific glutamate dehydrogenase of neurospora crassa, the secondary structures have been predicted from the rules of chou and fasman (chou, p.y., and fasman, g.d. (1979) biophys. j. 26, 367-384). comparison of these structures with those calculated for bovine glutamate dehydrogenase has shown that in the regions of homologous sequences containing identified functional regions, there is considerable homology of structure. from ...19806447152
novel features in the genetic code and codon reading patterns in neurospora crassa mitochondria based on sequences of six mitochondrial trnas.we report the sequences of neurospora crassa mitochondrial alanine, leucine(1), leucine(2), threonine, tryptophan, and valine trnas. on the basis of the anticodon sequences of these trnas and of a glutamine trna, whose sequence analysis is nearly complete, we infer the following: (i) the n. crassa mitochondrial trna species for alanine, leucine(2), threonine, and valine, amino acids that belong to four-codon families (gcn, cun, acn, and gun, respectively; n = u, c, a, or g) all contain an unmodi ...19806447871
eukaryotic dna segments capable of autonomous replication in yeast.a selective scheme is presented for isolating sequences capable of replicating autonomously in the yeast saccharomyces cerevisiae. yip5, a vector that contains the yeast gene ura3, does not transform a ura3 deletion mutant to ura+. hybrid yip5-escherichia coli dna molecules also fail to produce transformants. however, collections of molecular hybrids between yip5 and dna from any of six eukaryotes tested (s. cerevisiae, neurospora crassa, dictyostelium discoideum ceanhorabditis elegans, drosophi ...19806449009
the isolation and characterization of mutants defective in nitrate assimilation in neurospora crassa.the isolation and characterization of mutants altered for nitrate assimilation in neurospora crassa is described. the mutants isolated can be subdivided into five classes on the basis of growth test that correspond to the growth patterns of existing mutants at six distinct loci. mutants with growth characteristics like those of nit-2, nit-3 and nit-6 are assigned to those loci on the basis of noncomplementation and lack of recombination. mutants that, from their growth patterns, appear to lack t ...19806449399
the nucleotide sequence of phenylalanine trna from the cytoplasm of neurospora crassa.the phenylalanine trna from the cytoplasm of neurospora crassa has been purified and sequenced. the sequence is: pgcggguuuam2gcuca (n) gddgggagagcm22gpsicagacmugmaayapsim5cugaagm7gdm5cgugugtpsicgm1auccacacaaaccgcaccaoh. both in the nature of modified nucleotides which are present in this trna and in the overall sequence, this trna resembles more closely phenylalanine trnas of eukaryotic cytoplasm than those of prokaryotes. the sequence of this trna differs from those of the corresponding trnas o ...19806449691
a phenylalanine trna gene from neurospora crassa: conservation of secondary structure involving an intervening sequence.we have isolated and sequenced a trnaphe gene from neurospora crassa. hybridization analyses suggest that trnaphe is the only trna encoded on the cloned 5 kb dna fragment. the trnaphe gene contains an intervening sequence 16 nucleotides in length located one nucleotide 3' to the anticodon position. the trnaphe coding region of neurospora and yeast are 91% conserved, whereas their intervening sequences are only 50% identical. the pattern of sequence conservation is consistent with a proposed seco ...19806449692
evolutionary origins of vertebrate hormones: substances similar to mammalian insulins are native to unicellular eukaryotes.tetrahymena pyriformis, neurospora crassa, and aspergillus fumigatus that had been grown in simple defined media were extracted with acid ethanol by a classic method for recovering insulin from pancreas. after filtration of the extracts on sephadex g-50, distinct peaks of insulin immunoreactivity were recovered in the region typical of insulin. the gel-filtered material from the tetrahymena had reactivity in the pork insulin radioimmunoassay about equal to its reactivity in the insulin bioassay ...19806449704
biogenesis of glyoxysomes. synthesis and intracellular transfer of isocitrate lyase.biosynthesis of isocitrate lyase, a tetrameric enzyme of the glyoxysomal matrix, was studied in neurospora crassa, in which the formation of glyoxysomes was induced by a substitution of sucrose medium by acetate medium. 1. translation of neurospora mrna in reticulocyte lysates yields a product which has the same apparent molecular weight as the subunit of the functional enzyme. using n-formyl[35s]methionyl-trnafmet as a label, the translation product shows the same apparent size which indicates ...19806450681
purification, properties and synthesis of delta-aminolaevulinate dehydratase from neurospora crassa.delta-aminolaevulinate dehydratase, the second and rate-limiting enzyme of the haem-biosynthetic pathway, was purified 300-fold from induced cultures of neurospora crassa. the native enzyme has a mol.wt. of about 350000, whereas the salt-treated enzyme after incubation at 37 degrees c for 10 min has a mol.wt. of about 232000. the mol.wt. of the subunit is about 38000. antibodies to the purified enzyme were raised in rabbits. by using radiolabelling and immunoprecipitation techniques it was shown ...19806451221
kinetic properties of fatty acid synthetase of neurospora crassa.the kinetic properties of fatty acid synthetase, extracted from the mycelia of neurospora crassa, were studied to determine the role of the multi-enzyme complex in the regulation of long chain saturated fatty acid synthesis. acetyl-coa, a substrate, affects the fatty acid synthetase with increasing concentration in a normal michaelis-menton kinetic mode. malony-coa, another substrate, activates the synthetase in a homotropic manner up to 35 microm with a hill coefficient of 2.8. above that conce ...19806451784
mutagen sensitivities and mutator effects of mms-sensitive mutants in neurospora.7 mus (mutagen-sensitive) mutants of neurospora crassa, which are more sensitive to the toxic effects of mms (methyl methanesulfonate) than wild-type, were investigated for cross-sensitivities to other mutagens and inhibitors. these mutants have recently been mapped in 5 new genes, mus-7 to mus-11, and mutant alleles from each gene were checked for their effects on mutation frequencies. it was found that mutants in 3 of these 5 genes showed radiation-induced mutation frequencies similar to wild- ...19816451802
control of growth and of the nuclear division cycle in neurospora crassa. 19816452573
the nucleotide sequence at the 3'-end of neurospora crassa 25s-rrna and the location of a 5.8s-rrna binding site.the sequence of 110 nucleotides adjacent to the 3'-end of neurospora crassa 25s-rrna has been derived by chemical sequencing methods. sequences present between 40 and 85 nucleotides of the 3'-end were found to complement sequences at the 3'- and 5'-ends of 5.8s-rrna. interaction was shown to occur between 5.8s-rrna and a specific 3'-terminal fragment of 85 nucleotides derived from 25s-rrna. we have also demonstrated that the nucleotide sequence at the 3'-end of n. crassa 5.8s-rrna (-ucauuoh) is ...19816453330
nucleotide sequence of 5s ribosomal rna from aspergillus nidulans and neurospora crassa.the nucleotide sequences of 5s rrna molecules isolated from the cytosol and the mitochondria of the ascomycetes a. nidulans and n. crassa were determined by partial chemical cleavage of 3'-terminally labelled rna. the sequence identity of the cytosolic and mitochondrial rna preparations confirms the absence of mitochondrion-specific 5s rrna in these fungi. the sequences of the two organisms differ in 35 positions, and each sequence differs from yeast 5s rrna in 44 positions. both molecules conta ...19816453331
incorporation of photolabile azido fatty acid probes in neurospora crassa and saccharomyces cerevisiae. 19816453723
mitochondrial ribosome assembly in neurospora crassa. purification of the mitochondrially synthesized ribosomal protein, s-5.in neurospora, one mitochondrial ribosomal protein (s-5, mr = 52,000) is synthesized intramitochondrially and is presumably encoded by mitochondrial dna. we have developed a rapid method for the purification of s-5 which takes advantage of its high affinity for carboxymethyl-sepharose in the presence of 6 m urea. using this method, s-5, at purity greater than 95%, can be prepared by column chromatography in a single batch elution step. the amino acid composition of s-5 was determined. judged by ...19816453873
mutagenesis at the ad-3a and ad-3b loci haploid uv-sensitive strains of neurospora crassa. iii. comparison of dose-response curves for inactivation and mutation induced by gamma-rays.gamma-ray-induced inactivation and induction of mutations at the ad-3a and ad-3b loci of neurospora crassa have been compared among 6 different uv-sensitive strains and a standard wild-type strain. the 6 strains show varying degrees of sensitivity to gamma-ray-induced inactivation, with the relative sensitivity at 37% survival being uvs-6 greater than upr-1 greater than uvs-2 greater than uvs-3 greater than wild-type greater than uvs-5 greater than uvs-4. studies on the induction of ad-3 mutants ...19816454839
mutagenicity of fractions of cigarette smoke condensate in neurospora crassa and salmonella typhimurium.the mutagenicities of selected fractions of cigarette smoke condensate (csc) were studied in neurospora crassa for the presence of direct-acting mutagens. cscs from the university of kentucky reference cigarette 1r1 were assayed in a forward-mutation test at the adenine-3 (ad-3) region in resting conidia of a 2-component heterokaryon. direct-acting mutagenic activity was found in an enriched polycyclic aromatic hydrocarbons (epah) fraction and in a basic fraction (swain 5). no direct-acting muta ...19816454842
mutagenicity of cigarette smoke condensate in neurospora crassa.the mutagenicity of cigarette smoke condensate (csc) was investigated in neurospora crassa in the presence and absence of s9 mix prepared from aroclor-1254-induced rat liver. csc from the university of kentucky reference cigarette 1r1 was assayed in a forward-mutation test at the adenine-3 (ad-3) region in resting conidia of 2-component heterokaryons. in the absence of s9 mix, csc exhibited direct-acting mutagenicity. csc was also mutagenic in the presence of s9 mix, but higher doses were requir ...19816454843
genetic screening of compounds used in drug abuse treatment. ii. methadone.several compounds used clinically in drug abuse therapy were evaluated for genetic activity in a series of in vivo and in vitro assays. the second report in this series describes the results for one of these compounds, methadone. a previous report described the results for naltrexone. methadone is a synthetic narcotic analgesic used as a substitute for heroin in drug detoxification programs. methadone demonstrated weak activity in the e. coli dna repair system and in the neurospora crassa and th ...19816455283
genetic screening of compounds used in drug abuse treatment. iii. laam.several compounds used clinically in drug abuse therapy were evaluated for genetic activity in a series of in vivo and in vitro assays. this third report in the series describes the results for one of these compounds, laam (l-alpha-acetyl methadol). previous reports described results from a three phase testing program for naltrexone and methadone. laam is related chemically to the narcotic analgesic oxymorphine, and is related chemically to a narcotic antagonist, naloxone. laam exhibited genetic ...19816455284
effect of regulatory mutations of sulphur metabolism on the levels of cysteine- and homocysteine-synthesizing enzymes in neurospora crassa.1. regulation of four enzymes involved in cysteine and homocysteine synthesis, i.e. cysteine synthase (ec 4.2.99.8), homocysteine synthase (ec 4.1.99.10), cystathionine beta-synthase (ec 2.1.22) and gamma-cystathionase (ec 4.4.1.1) was studied in the wild type and sulphur regulatory mutants of neurospora crassa. 2. homocysteine synthase and cystathionine beta-synthase were found to be regulatory enzymes but only the former is under control of the cys-3 - scon system regulating several enzymes of ...19806455895
studies on myo-inositol-1-phosphate from lilium longiflorum pollen, neurospora crassa and bovine testis. further evidence that a classical aldolase step is not utilized. 19816456770
neurospora crassa nad(p)h-nitrite reductase. studies on its composition and structure.neurospora crassa nitrite reductase (mr = 290,000) catalyzes the nad(p)h-dependent 6-electron reduction of nitrite to ammonia via flavin and siroheme prosthetic groups. homogeneous n. crassa nitrite reductase has been prepared employing conventional purification methods followed by affinity chromatography on blue dextran-sepharose 4b. sodium dodecyl sulfate-polyacrylamide gel electrophoresis of homogeneous nitrite reductase reveals a single subunit band of mr = 140,000. isoelectric focusing of d ...19816457037
folylpoly-gamma-glutamate synthesis by bacteria and mammalian cells.the purification and properties of folylpolyglutamate synthetase from corynebacterium sp, and some properties of partially purified enzyme from lactobacillus casei, streptococcus faecalis, neurospora crassa, pig liver, and chinese hamster ovary cells, are described. the corynebacterium enzyme catalyzes a mgatp-dependent addition of glutamate to a variety of reduced pteroate and pteroylmono-, di-, and triglutamate substrates, with the concomitant production of mgadp and phosphate. although glutam ...19816458762
the cell-free protein synthesis system from the 'slime' mutant of neurospora crassa. preparation and characterisation of importance of 7-methylguanosine for translation of viral and cellular mrnas.a simple procedure for preparation of a cell-free protein synthesis system (23000 x g supernatant) from the protoplast-like 'slime' mutant of neurospora crassa is described. a variety of messenger rnas of viral and cellular origin could be efficiently and faithfully translated in this system into proteins with mr as large as 180000. the importance of the 7-methylguanosine cap for mrna translation in the neurospora system was studied in detail using the cap analogs and chemically decapped messeng ...19816459928
role of lipids in the neurospora crassa membrane: iv. biochemical and electrophysiological changes caused by growth on phytanic acid.neurospora crassa strain cel, which is deficient in fatty acid synthesis, was grown with phytanic acid supplementation. the temperature dependence of membrane potential is increased by growth on phytanic acid. a temperature change of 40 degrees c produces a change of 184 mv in phytanic acid-grown cells as compared to a 50 mv change for cel grown on palmitic acid or wild-type. membrane resistance (measured as dc input resistance) of phytanic acid-grown cells did not differ from cel grown on palmi ...19826460106
biochemical analysis of mutants defective in nitrate assimilation in neurospora crassa: evidence for autogenous control by nitrate reductase.a biochemical analysis of mutants altered for nitrate assimilation in neurospora crassa is described. mutant alleles at each of the nine nit (nitrate-nonutilizing) loci were assayed for nitrite reductase activity, for three partial activities of nitrate reductase, and for nitrite reductase activity. in each case, the enzyme deficiency was consistent with data obtained from growth tests and complementation tests in previous studies. the mutant strains at these nit loci were also examined for alte ...19816460156
mutagenesis at the ad-3a and ad-3b loci in haploid uv-sensitive strains of neurospora crassa. v. comparison of dose--response curves of single- and double-mutant strains with wild-type.the interactions of mutant alleles that individually confer radiation sensitivity in neurospora crassa are being studied with regard to their effects on radiation-induced inactivation and forward-mutation induction at the ad-3 loci. this paper reports attempts to construct 3 double-mutant strains containing the following pair-wise combinations of repair-deficient mutants: upr-1,uvs-2; uvs-2,uvs-6; and uvs-3,uvs-6. the double-mutant strain with the 2 excision-repair-deficient mutants upr-1 and uv ...19816460180
solubility of (1 leads to 3)-beta-d/(1 leads to 6)-beta-d-glucan in fungal walls: importance of presumed linkage between glucan and chitin.in saccharomyces cerevisiae, neurospora crassa, aspergillus nidulans and coprinus cinereus most of the alkali-insoluble (1 leads to 3)-beta-d/(1 leads to 6)-beta-d-glucan of the wall can be extracted with dimethyl sulphoxide. the same fraction, and in saccharomyces cerevisiae a small additional fraction, can be extracted by a destructive procedure involving 40% naoh at 100 degrees c. the small fraction of the glucan which resists this treatment becomes soluble after a subsequent treatment with h ...19816460846
use of four short-term tests to evaluate the mutagenicity of municipal water.some ways in which four short-term tests may be used to evaluate the mutagenicity of drinking water were explored by testing raw and treated water from lake bloomington, which serves the town of bloomington, illinois (population, 44,000). the water was collected from february 1976 to october 1977 and was concentrated by evaporation or by use of xad-2 resin. the water was tested for the ability to induce reverse mutation in a prokaryote, salmonella typhimurium; forward mutation in a mold, neurosp ...19826460874
estimation of deuteration levels in whole cells and cellular proteins by 1h n.m.r. spectroscopy and neutron scattering.methods using conventional fourier transform 1h n.m.r. spectroscopy at 250 mhz for the determination of the overall deuteration levels of cells cultured in media containing 2h2o or deuterated carbon sources are described. these were developed for escherichia coli as a model, and extended to neurospora crassa hyphae and mouse myeloma cells p815. the results were investigated by 1h n.m.r. and neutron scattering measurements on deuterated proteins that were obtained from e. coli. it is concluded th ...19816461329
induction of acyl coenzyme a synthetase and hydroxyacyl coenzyme a dehydrogenase during fatty acid degradation in neurospora crassa.neurospora crassa is able to use long-chain fatty acids as the sole carbon and energy source. after growth on oleate there was nearly a 10-fold induction of the acyl coenzyme a (coa) synthetase and a fivefold increase in the activity of the 3-hydroxyacyl-coa dehydrogenase. there was a slight induction of the enoyl-coa hydratase and 3-ketoacyl-coa thiolase, but no apparent induction of the flavin-linked acyl-coa dehydrogenase. these noncoordinate changes in the fatty acid degradation enzymes sugg ...19826461637
[problems concerned with microbial mutagenicity tests].mutagenicity tests using microorganisms are the salmonella-typhimurium test (ames), the test with a polymerase a deficient escherichia coli, the saccharomyces cerevisiae d3 system and the neurospora crassa test. differences in results of ames-test may be due to differences preserving the tester strains, the choice of solvent and the dosage of the test compounds, last not least is the production of s-9. the polymerase a deficient escherichia coli system seems easier to handle than the saccharomyc ...19826461834
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
the biosynthesis of brominated pyrrolnitrin derivatives by pseudomonas aureofaciens.the mutant strain acn of pseudomonas aureofaciens atcc 15926 produces several bromo derivatives of pyrrolnitrin. five brominated amino- and three brominated nitrophenyl pyrrole compounds could be isolated, and their structures were established by 1h nmr, uv and mass spectroscopy. the isolated amino compounds showed no biological activity; the nitro derivatives inhibited the growth of neurospora crassa atcc 9276, though not as effective as pyrrolnitrin itself. 2-carboxy-4-(2-amino-3-bromophenyl)p ...19836662814
molybdenum cofactor from the cytoplasmic membrane of proteus mirabilis.molybdenum cofactor was extracted from membranes of proteus mirabilis by three methods: acidification, heat treatment and heat treatment in the presence of sodium-dodecylsulphate (sds). extracts prepared by the latter method contained the highest concentration of molybdenum cofactor. in these extracts molybdenum cofactor was present in a low molecular weight form. it could not penetrate an ym-2 membrane during ultrafiltration suggesting a molecular weight above 1000. during aerobic incubation of ...19826763509
active polypeptide fragments common to prokaryotic, eukaryotic, and mitochondrial dna polymerases.with a procedure that allows the renaturation of the dna polymerase catalytic activity in situ after sds-polyacrylamide gel electrophoresis, we have compared the active polypeptides present in extracts from organisms covering a wide evolutionary range from prokaryotes to eukaryotes, namely: escherichia coli, oryza sativa, daucus carota , neurospora crassa, dictyostelium discoideum, saccharomyces cerevisiae, ceratitis capitata, leucophaea maderae , xenopus laevis, rat tissues and human lymphoblas ...19826765191
construction and characterization of salmonella typhimurium strains that accumulate and excrete alpha- and beta-isopropylmalate.two salmonella typhimurium strains, which could be used as sources for the leucine biosynthetic intermediates alpha- and beta-isopropylmalate were constructed by a series of p22-mediated transductions. one strain, jk527 [flr-19 leua2010 delta(leud-ara)798 fol-162], accumulated and excreted alpha-isopropylmalate, whereas the second strain, jk553 (flr-19 leua2010 leub698), accumulated and excreted alpha- and beta-isopropylmalate. the yield of alpha-isopropylmalate isolated from the culture medium ...19806991477
identification of the molybdenum cofactor in chlorate-resistant mutants of escherichia coli.experiments were performed to determine whether defects in molybdenum cofactor metabolism were responsible for the pleiotropic loss of the molybdoenzymes nitrate reductase and formate dehydrogenase in chl mutants of escherichia coli. in wild-type e. coli, molybdenum cofactor activity was present in both the soluble and membrane-associated fractions when the cells were grown either aerobically or anaerobically, with and without nitrate. molybdenum cofactor in the soluble fraction decreased when t ...19817026535
the influence of growth conditions on the synthesis of molybdenum cofactor in proteins mirabilis.cell-free extracts of proteus mirabilis were able to reconstitute nadph-dependent assimilatory nitrate reductase in crude extracts of the neurospora crassa mutant strain nit-1, lacking molybdenum cofactor. molybdenum cofactor was formed in the cytoplasm of the bacterium even in the presence of oxygen during growth though under these conditions no molybdo enzymes are formed. as a consequence no cofactor could be released by acid treatment from membranes of cells growth aerobically. the amount of ...19817030254
pyruvate dehydrogenase complex from baker's yeast. 1. purification and some kinetic and regulatory properties.pyruvate dehydrogenase complex, for the first time, was highly purified from commercial baker's yeast (saccharomyces cerevisiae). proteolytic degradation was prevented by the inclusion of the protease inhibitors pepstatin a, leupeptin, and phenylmethanesulfonyl fluoride during the enzyme purification. the yield from 1 kg of pressed yeast was about 15--20 mg enzyme with a specific activity of 17--30 u/mg. most of the kinetic and regulatory properties of the yeast enzyme were found similar to thos ...19817030740
mutagenicity of oil-shale ash.3 oil-shale ash samples were extracted with solvents and analyzed for mutagenicity with a number of tests systems. in salmonella typhimurium, the ash extracts were highly mutagenic with the ames his reversion and the ara-resistant systems. mutation induction by the ash in salmonella was independent of metabolic activation and was of the frameshift type. these ash extracts showed a substantial killing effect, but failed to induced ad-3 reversion in neurospora crassa, gene conversion and mitotic c ...19827035913
nuclear localization of aspartate transcabamoylase in saccharomyces cerevisiae.the cytochemical technique using the in situ precipitation of orthophosphate ions liberated specifically by the aspartate carbamoyltransferase (atcase) (ec 2.1.3.2) reaction indicated that in saccharomyces cerevisiae this enzyme is confined to the nucleus. this observation is in accordance with the result reported by bernhardt and davis (1972), proc. natl. acad. sci. u. s. a. 69:1868-1872) on neurospora crassa. the nuclear compartmentation was also observed in a mutant strain lacking proteinase ...19827045137
genetical and biochemical aspects of quinate breakdown in the filamentous fungus aspergillus nidulans.in the ascomycetous fungus aspergillus nidulans, the expression of two inducible, contiguous or closely linked genes (qutb and qutc) which encode enzymes for quinate breakdown to protocatechuate, appears to be controlled by the product of a tightly linked third genet (quta). the qut gene cluster locates on chromosome viii. the catalytic steps required for this conversion are dehydrogenase, dehydroquinase, and dehydratase, and these activities are induced by the presence of quinate in a similar m ...19827049157
[refractory properties of cardiac tissue at fast sodium current decrease. comparison of atrium and ventricle].investigation of the membrane potential of neurospora crassa mycelial cells was carried out by standard microelectrode technique. the resting potential is equal to--156 +/- 11 mv (negative inside the cell). the light of the spectrum blue-violet zone causes transient hyperpolarization of the cell membrane reaching --38 +/- 5 mv after 25 minutes of illumination.19827138947
two classes of replicating molecules of adenovirus type 2 dna.replicating dna of human adenovirus type 2, identified as partly single-stranded viral dna in which [3h]thymidine is readily incorporated, was found to be separated into two fractions by chromatography on hydroxyapatite. whereas one of the these fractions was eluted with 180 mm phosphate, the other one was eluted at the same concentration, 240 mm, as fully double-stranded dna. the physical properties of the 180 and 240 mm fractions, in particular their buoyant densities in solutions of cscl and ...19817248295
physicochemical characterization of the ribosomal rna species of the mollusca. molecular weight, integrity and secondary-structure features of the rna of the large and small ribosomal subunits.1. the rrna species of the cephalopoda octopus vulgaris and loligo vulgaris were found to have unexpectedly high sedimentation coefficients and molecular weights. in 0.1 m-nacl the l-rrna (rna from large ribosomal subunit) has the same s20 value as the l-rrna of the mammals (30.7s), whereas the s-rrna (rna from small ribosomal subunit) sediments at a faster rate (20.1s) than the s-rrna of both the mammals and the fungi (neurospora crassa) (17.5s). the molecular weights of the l-rrna were determi ...19807458915
a human homolog of the mitochondrial protein import receptor mom19 can assemble with the yeast mitochondrial receptor complex.import of preproteins into mitochondria requires transport machineries in both mitochondrial membranes that have been characterized in saccharomyces cerevisiae and neurospora crassa. by cdna analysis, we identified a human protein of 16 kda with significant overall homology to the fungal mitochondrial import receptor mom19, including the three typical characteristics: a hydrophobic n-terminal segment, a tetratrico peptide motif in the middle and a negatively charged c-terminus. the human mom19 h ...19957498524
palindromic repeated sequences (prss) in the mitochondrial genome of rice: evidence for their insertion after divergence of the genus oryza from the other gramineae.we have identified a family of small repeated sequences (from 60 to 66 bp in length) in the mitochondrial genome of rice (oryza sativa cv. nipponbare). there are at least ten copies of these sequences and they are distributed throughout the mitochondrial genome. each is potentially capable of forming a stem-and-loop structure and we have designated them prss (palindromic repeated sequences). their features are reminiscent of the small dispersed repeats in the mitochondrial dna (mtdna) of some lo ...19947509206
x-ray-induced specific-locus mutations in the ad-3 region of two-component heterokaryons of neurospora crassa. xii. analysis of multiple-locus ad-3 mutations reveals a nonrandom distribution of the separate sites of recessive lethal damage throughout the genome.previous studies on x-ray-induced adenine-3 mutations induced in heterokaryon 12 of neurospora crassa showed that they consisted of gene/point mutations, multilocus deletion mutations, and 3 different genotypic classes of multiple-locus mutations (designated [-3]ir + rlcl, ad-3r + rlcl, and ad-3r + rl). in the present paper, multiple-locus mutations consisting of gene/point mutations at the ad-3a or the ad-3b locus with sites of recessive lethal damage closely linked to the ad-3 region (designat ...19947513795
forward-mutation tests on the antitumor agent icr-170 in neurospora crassa demonstrate that it induces gene/point mutations in the ad-3 region and an exceptionally high frequency of multiple-locus ad-3 mutations with closely linked sites of recessive lethal damage.the mutagenicity of the antitumor agent icr-170 (2-methoxy-6-chloro-9-[(ethyl-2-chloroethyl)amino propylamino] acridine dihydrochloride) in the adenine-3 (ad-3) region was studied with a two-component heterokaryon (h-12) of neurospora crassa. the objective was to characterize the genetic damage produced by this acridine nitrogen mustard derivative to determine in a lower eukaryotic organism the basis for its potent activity against ascites tumors in mice. as in higher eukaryotes, specific-locus ...19947523879
peptide-specific antibodies as probes of the topography of the voltage-gated channel in the mitochondrial outer membrane of neurospora crassa.the voltage-dependent anion-selective channel (vdac) in mitochondrial outer membranes is formed by a polypeptide (m(r) 31,000) coded by a nuclear gene whose cdna sequence is known for several organisms. antibodies have been raised against synthetic peptides corresponding to four different regions in the predicted sequence of the vdac polypeptide of the fungus neurospora crassa (residues 1-20, amino terminus; 195-210, 251-268, and 272-283, carboxyl terminus). specificity of the antibodies has bee ...19957542652
homology cloning of gtp-cyclohydrolase i from various unrelated eukaryotes by reverse-transcription polymerase chain reaction using a general set of degenerate primers.gtp-cyclohydrolase i is the primary enzyme of tetrahydrobiopterin and folic acid biosynthesis. cdna fragments of gtp-cyclohydrolase i were obtained from rainbow trout, chicken, the fungi neurospora crassa, phycomyces blakesleeanus and saccharomyces cerevisiae, the cellular slime mold dictyostelium discoideum, the phytoflagellate euglena gracilis and the higher plant mucuna hassjo using primers specific for conserved regions of the open reading frame and the reverse transcription polymerase chain ...19957542887
phylogenetic ordinal placement based on rdna sequences of the freshwater genera ophioceras and pseudohalonectria.the ordinal placement of two closely related freshwater genera, ophioceras and pseudohalonectria, was assessed by using phylogenetic analysis of morphological characters, partial sequences of the large subunit ribosomal dna and restriction site variations in the internal transcribed spacer (its). the two genera have some morphological features that are used to define taxa in both the sordariales and diaporthales, and, hence, their phylogenetic relationships are unclear. equally weighted analyses ...19957553269
sequence of the met-10+ locus of neurospora crassa: homology to a sequence of unknown function in saccharomyces cerevisiae chromosome 8.we have determined the sequence of the neurospora crassa met-10+ gene and its flanking regions, and have isolated and analyzed cdna clones for this region. we have identified two closely linked genes transcribed in the same orientation. the met-10+ gene is the downstream gene; an open reading frame (orf) derived from five exons encodes a 475-amino-acid protein. the deduced protein lacks similarity to other characterized proteins. however, it exhibits a strong similarity to the product of an orf ...19957557397
escherichia coli chorismate synthase catalyzes the conversion of (6s)-6-fluoro-5-enolpyruvylshikimate-3-phosphate to 6-fluorochorismate. implications for the enzyme mechanism and the antimicrobial action of (6s)-6-fluoroshikimate.chorismate synthase catalyzes the conversion of 5-enolpyruvylshikimate-3-phosphate to chorismate. it is the seventh enzyme of the shikimate pathway, which is responsible for the biosynthesis of aromatic metabolites from glucose. the chorismate synthase reaction involves a 1,4-elimination with unusual anti-stereochemistry and requires a reduced flavin cofactor. the substrate analogue (6s)-6-fluoro-5-enolpyruvylshikimate-3-phosphate is a competitive inhibitor of neurospora crassa chorismate syntha ...19957559411
molecular characterization of the proline-1 (pro-1) locus of neurospora crassa, which encodes delta 1-pyrroline-5-carboxylate reductase.delta 1-pyrroline-5-carboxylate reductase (p5cr; [l-proline: nad(p+) 5-oxidoreductase]; ec 1.5.1.2) catalyzes the final step in proline biosynthesis. we have shown that the proline-1 (pro-1) locus of neurospora crassa encodes p5cr. the pro-1 gene was localized to a 3.2 kb region by complementation of (restoration of proline-independent growth to) a proline auxotroph carrying a recessive mutation at the pro-1 locus. the nucleotide sequence of this 3.2 kb region contains an open reading frame with ...19957565596
translational regulation in response to changes in amino acid availability in neurospora crassa.we examined the regulation of neurospora crassa arg-2 and cpc-1 in response to amino acid availability.arg-2 encodes the small subunit of arginine-specific carbamoyl phosphate synthetase; it is subject to unique negative regulation by arg and is positively regulated in response to limitation for many different amino acids through a mechanism known as cross-pathway control. cpc-1 specifies a transcriptional activator important for crosspathway control. expression of these genes was compared with ...19957565672
immunological evidence for accumulation of two high-molecular-weight (104 and 90 kda) hsps in response to different stresses in rice and in response to high temperature stress in diverse plant genera.rice seedlings accumulate stainable amounts of the 104 and 90 kda polypeptides in response to high temperature stress. we have purified and raised highly specific polyclonal antisera against both of these polypeptides. in western blotting experiments, we find that these proteins are accumulated to different extents in rice seedlings subjected to salinity (nacl), water stress, low-temperature stress and exogenous abscisic acid application. these proteins also accumulated when rice seedlings grown ...19957579180
design of an object-oriented database for reverse genetics.we present the design of an object-oriented database system for reverse genetics applications. such a database will encapsulate not only the data in the genetic and physical maps, but also the methods used to create the maps as well as methods to link them to other databases, such as genbank, pir, and medline. the purpose of this database is to provide the fungal genetics community with an electronic tool for identifying the biochemical function of any dna fragment in the database--electronic re ...19937584341
nucleotide sequence and characterization of the large mitochondrial rrna gene of penicillium urticae, and its comparison with those of other filamentous fungi.the nucleotide sequence of a large rrna gene and its flanking regions in cloned fragments of mitochondrial dna from a patulin producer, penicillium urticae nrrl2159a, was determined by dideoxy sequencing, and the 5' end and intron-exon border of the 1-rrna gene were determined by primer extension analysis and rna sequencing, respectively. in addition to the extensive sequence homology of the 3' end of the p. urticae mt 1-rrna gene with those of aspergillus nidulans and neurospora crassa, the p. ...19957592555
molecular cloning and functional expression of neurospora deoxyhypusine synthase cdna and identification of yeast deoxyhypusine synthase cdna.deoxyhypusine synthase catalyzes the formation of deoxyhypusine residue on the eif-5a precursor using spermidine as the substrate. we have purified deoxyhypusine synthase from neurospora crassa to apparent homogeneity (tao, y., and chen, k. y. (1995) j. biol. chem. 270, 383-386). we have now cloned and characterized the deoxyhypusine synthase cdna using a reverse genetic approach. conceptual translation of the nucleotide sequence of the cloned 1258-base pair cdna revealed an open reading frame c ...19957592594
assembly of the preprotein receptor mom72/mas70 into the protein import complex of the outer membrane of mitochondria.membrane integration and assembly of mom72 from neurospora crassa and its yeast homolog mas70 was studied with isolated mitochondria. after synthesis in vitro, the precursors of mom72/mas70 are tightly folded and expose only their n-terminal amino acid residues comprising the targeting and the membrane anchor domain. insertion of the protein into the mitochondrial outer membrane (mom) occurs in a time- and temperature-dependent manner and is stimulated by atp. mom72/mas70 is then assembled into ...19957592965
the negative-acting nmr regulatory protein of neurospora crassa binds to and inhibits the dna-binding activity of the positive-acting nitrogen regulatory protein nit2.structural genes of the nitrogen regulatory circuit of the filamentous fungus neurospora crassa are under the control of both positive and negative regulatory proteins. nit2, the major positive-acting nitrogen regulatory protein, activates the expression of structural genes within the nitrogen circuit. nit2 binds to upstream activation sites which contain at least two gata core elements in the promoter regions of the nitrogen-controlled structural genes, and activates their transcription, possib ...19957612627
(1,3)-beta-d-glucan synthase activity in mycelial and cell wall-less phenotypes of the fz, sg, os-1 ("slime") mutant strain of neurospora crassa.the cell wall-less fz, sg, os-1 ("slime") triple mutant of neurospora crassa lacks (1,3)-beta-d-glucan synthase activity. fz, sg, os-1 segregants from slime x wild-type crosses initially germinate as a plasmodium (slime-like), but develop hyphae in a few hours and acquire a stable mycelial phenotype (mycelial intermediate). the cell wall-less phenotype (stable slime) can be reisolated from mycelial intermediates by filtration-enrichment selection in medium of high osmolarity. pairs of mycelial i ...19957614368
nuclear dna-binding proteins which recognize the intergenic control region of penicillin biosynthetic genes.the biosynthesis of penicillin, a secondary metabolite produced by penicillium chrysogenum, is subject to sophisticated genetic and metabolic regulation. the structural genes, pcbc and pcbab, which encode two of the penicillin biosynthetic enzymes are separated by a 1.16-kb intergenic region and transcribed divergently from one another. to identify and characterize nuclear proteins which interact with the pcbab-pcbc intergenic promoter region, crude and partially purified nuclear extracts were u ...19957614558
isolation of the vma-4 gene encoding the 26 kda subunit of the neurospora crassa vacuolar atpase.we have isolated the vma-4 gene, which encodes a 25,746 dalton subunit of the vacuolar atpase, from neurospora crassa. the gene contains two introns and was mapped to the left arm of linkage group i. comparison of the predicted amino acid sequence with homologous proteins from saccharomyces cerevisiae, manduca sexta, and bos taurus showed only 25% sequence identity. however, computer-assisted predictions of secondary structures gave similar results for all four proteins. analysis of the sequence ...19957619848
conservative sorting of f0-atpase subunit 9: export from matrix requires delta ph across inner membrane and matrix atp.in an attempt to understand the mechanisms of sorting of mitochondrial inner membrane proteins, we have analyzed the import of subunit 9 of the mitochondrial f1f0-atpase (su9) from neurospora crassa, an integral inner membrane protein. a chimeric protein was used consisting of the presequence and the first transmembrane domain of su9 fused to mouse dihydrofolate reductase (presu9(1-112)-dhfr). this protein attains the correct topology across the inner membrane (nout-cin) following import. the tr ...19957628445
isolation of a functional homolog of the cell cycle-specific nima protein kinase of aspergillus nidulans and functional analysis of conserved residues.to investigate the degree of conservation of the cell cycle-specific nima protein kinase of aspergillus nidulans, and to help direct its functional analysis, we cloned a homolog (designated nim-1) from neurospora crassa. over the catalytic domain nim-1 is 75% identical to nima, but overall the identity drops to 52%. nim-1 was able to functionally complement nima5 in a. nidulans. mutational analysis of potential activating phosphorylation sites found in nima, nim-1, and related protein kinases wa ...19957629122
purification, n-terminal amino acid sequence and partial characterization of a cu,zn superoxide dismutase from the pathogenic fungus aspergillus fumigatus.a superoxide dismutase (sod) has been purified to homogeneity from the fungal pathogen aspergillus fumigatus using a combination of cell homogenization, isoelectric focusing and gel filtration fplc. the n-terminal amino acid sequence of the purified enzyme demonstrated substantial homology to known cu,zn superoxide dismutases for a range of organisms, including neurospora crassa and saccharomyces cerevisiae. the enzyme subunit has a pi of 5.9, a relative molecular mass of 19 kda and a spectral a ...19957633574
the cellulase complex of neurospora crassa: cbh-1 cloning, sequencing and homologies.we describe the isolation, cloning and sequencing of the cellobiohydrolase 1 (ec 3.2.1.91)-encoding gene (cbh-1) of neurospora crassa. the nucleotide and amino-acid sequences have high homology with the cbh-1 of trichoderma reesei, humicola grisea and phanerochaete chrysosporium, with clear signal, catalytic, hinge and substrate-binding domains in that order.19957642129
the cpc-2 gene of neurospora crassa encodes a protein entirely composed of wd-repeat segments that is involved in general amino acid control and female fertility.phenotypic and molecular studies of the mutation u142 indicate that the cpc-2+ gene is required to activate general amino acid control under conditions of amino acid limitation in the vegetative growth phase, and for formation of protoperithecia in preparation for the sexual phase of the life cycle of neurospora crassa. the cpc-2 gene was cloned by complementation of the cpc-2 mutation in a his-2ts bradytrophic background. genomic and cdna sequence analysis indicated a 1636 bp long open reading ...19957651339
the aspergillus uvsh gene encodes a product homologous to yeast rad18 and neurospora uvs-2.the uvsh dna repair gene of aspergillus nidulans has been cloned by complementation of the uvsh77 mutation with a cosmid library containing genomic dna inserts from a wild-type strain. methylmethane sulfonate (mms)-resistant transformants were obtained on medium containing 0.01% mms, to which uvsh mutants exhibit high sensitivity. retransformation of uvsh77 mutants with the rescued cosmids from the mms-resistant transformants resulted in restoration of both uv and mms resistance to wild-type lev ...19957651340
the fusarium solani gene encoding kievitone hydratase, a secreted enzyme that catalyzes detoxification of a bean phytoalexin.among the antimicrobial phytoalexins produced by phaseolus vulgaris (french bean) is the prenylated isoflavonoid, kievitone. the bean pathogen, fusarium solani f. sp. phaseoli, secretes a glycoenzyme, kievitone hydratase (ec 4.2.1.95), which catalyzes conversion of kievitone to a less toxic metabolite. among f. solani strains, those that are highly virulent to p. vulgaris also produce kievitone hydratase constitutively, suggesting that the enzyme is a virulence factor. based on the n-terminal am ...19957655061
cloning and characterization of a heterologously expressed bifunctional chorismate synthase/flavin reductase from neurospora crassa.activities of all chorismate synthases (cs) so far analyzed are absolutely dependent upon reduced flavin. for monofunctional css, which represent the only class of css that have yet been cloned, the flavin must be reduced either (photo-)chemically or by a separable flavin reductase (fr) for in vitro activity. neurospora crassa cs, in contrast, possesses an intrinsic fr activity and represents the only firmly established member of a bifunctional class of css. to better understand this bifunctiona ...19957657620
cloning and expression of human deoxyhypusine synthase cdna. structure-function studies with the recombinant enzyme and mutant proteins.deoxyhypusine synthase catalyzes the first step in the post-translational formation of hypusine (n epsilon-(4-amino-2-hydroxybutyl)lysine). cdna clones encoding deoxyhypusine synthase were isolated from a human hela cell library. full-length cdna clones encoding a 369-amino acid protein (calculated molecular mass of 40,970 da) and a shorter cdna clone that would potentially encode a protein with an internal deletion of 56 amino acids (asp262-ser317) were isolated. the deduced amino acid sequence ...19957673224
characterization of sfp2, a putative sulfate permease gene of saccharomyces cerevisiae.the sfp2 gene of saccharomyces cerevisiae has been characterized. the deduced amino acid sequence contained twelve highly hydrophobic domains and showed 50, 47, 44 and 48% homologies to neurospora crassa sulfate permease ii (cys14), soybean gmak170 nodulin, human colon mucosa protein (dra) and a putative open reading frame (orf) downstream of escherichia coli prs (phosphoribosyl pyrophosphatate synthetase) gene, respectively, in the aligned regions. cells lacking sfp2 were viable and displayed n ...19957677785
comparison of the spectra of genetic damage in n4-hydroxycytidine-induced ad-3 mutations between nucleotide excision repair-proficient and -deficient heterokaryons of neurospora crassa.a comparison has been made of the mutagenic effects of n4-hydroxycytidine (hc) in the adenine-3 (ad-3) region of two-component heterokaryons of neurospora crassa: nucleotide excision repair-proficient (uvs-2+/uvs-2+) heterokaryon 12 (h-12) and nucleotide excision repair-deficient (uvs-2/uvs-2) heterokaryon 59 (h-59). hc was found to produce mutations predominantly, if not exclusively, by at to gc base-pair transitions in escherichia coli strain k12 by janion and glickman (1980, mutation res., 72 ...19937678887
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