TitleAbstractYear(sorted ascending)
the imported preprotein of the proteolipid subunit of the mitochondrial atp synthase from neurospora crassa. molecular cloning and sequencing of the mrna.the proteolipid subunit of the mitochondrial atp synthase from neurospora crassa is an extremely hydrophobic protein of 81 amino acid residues, which is imported into mitochondria as a precursor of mol. wt. 15 000. the primary structure of the imported form has now been determined by isolating and analyzing cdna clones of the preproteolipid mrna. an initial cdna clone was identified by hybridizing total polyadenylated rna to pooled cdna recombinant plasmids from an ordered clone bank and subsequ ...19826329691
mitochondrial variants of neurospora intermedia from nature.from a sample of 122 natural isolates of neurospora intermedia collected recently from around the world, five variants had erratic stop-start growth patterns reminiscent of the phenotype of "stopper" laboratory extranuclear mutants of neurospora crassa. like laboratory isolated mutants, the natural "stopper" variants were sterile as protoperithecial parents and transmitted the variant growth phenotypes very inefficiently, if at all, as male parents. heterokaryon tests could not be made because o ...19826303535
structure and organization of trna, rrna, and protein genes in neurospora crassa mitochondria.our studies on neurospora crassa mitochondria have included sequence analysis of trnas, mapping and cloning of the trna, rrna, and protein genes and the dna sequence analysis of these genes. results from trna sequence analyses explain how the mitochondrial protein synthesizing system can function with a much smaller number of trnas than other systems. mapping studies have shown that the two rrna genes and almost all of the trna genes are clustered onto a third of the mitochondrial genome. the tw ...19826300028
transformation of neurospora crassa utilizing recombinant plasmid dna. 19826279089
gene cloning in neurospora crassa. 19826276097
separation of neurospora crassa myo-inositol-1-phosphate synthase from glucose-6-phosphate dehydrogenase by affinity chromatography.the purification of neurospora crassa myo-inositol-1-phosphate synthase (ec was studied by affinity chromatography using the substrate (glucose-6-phosphate), the inhibitor (pyrophosphate), the coenzyme (nad+) and the coenzyme analogues (5'amp and cibacron blue f3g-a) of the enzyme as adsorbents attached to agarose gel. myo-inositol-1-phosphate synthase could be separated completely from the contaminating substance, glucose-6-phosphate dehydrogenase (ec, on blue sepharose cl-6b ...19826214775
nitrogen metabolite repression of fluoropyrimidine resistance and pyrimidine uptake in neurospora crassa.wild type neurospora crassa was shown to be more resistant to 5-fluoro-uracil, 5-fluoro-uridine and 5-fluoro-2'-deoxyuridine in the presence of ammonia than in its absence. this differential resistance may in part be accounted for by the observation that both uracil and uridine uptake in germinating conidia is under nitrogen metabolite regulation. the uptake of uracil and uridine is increased on poor nitrogen sources in wild type, is unaffected by nitrogen source in a nit-2 mutant strain while a ...19826213838
radioautographic detection of macromolecules involved in the synthesis of neurospora crassa cell wall. 19826213305
biosynthesis of glyoxysomal enzymes in neurospora crassa. 19826212015
two types of microbodies in neurospora crassa. 19826212014
postreplication repair in neurospora crassa.changes in the molecular weight of nascent dna made after ultraviolet (uv) irradiation have been studied in the excision-defective neurospora mutant uvs-2 using isotopic pulse labeling, alkaline gradient centrifugation and alkaline filter elution. both the size of nascent dna and the rate of incorporation of label into dna was reduced by uv light in a dose dependent manner. however, this dna repair mutant did recover the ability to synthesize control-like high molecular weight dna 3 hours after ...19826211589
relationship between the composition of phospholipids and respiratory activity of choline-deficient mutants of neurospora crassa.phosphatidylcholine is one of the most frequent phospholipid components of the inner mitochondrial membrane of neurospora crassa. quantitative analysis of phosphlipids of the wild strain of neurospora crassa and of its two cho mutants showed that these strains did not significantly differ in the content of phosphatidylcholine. mutants cultivated in a medium without choline contained, as compared with the wild strain, an increased amount of phosphatidylserine and a decreased quantity of phosphati ...19826211400
regulation of l-amino acid oxidase and of d-amino acid oxidase in neurospora crassa.neurospora crassa possesses an inducible l-amino acid oxidase that is expressed only when cells are derepressed for nitrogen in the presence of an amino acid. enzyme synthesis requires both induction by an amino acid and simultaneous nitrogen catabolite derepression. carbon limition in the presence of an amino acid does not permit induction of l-amino acid oxidase. the nit-2 gene is a major regulatory locus which is believed to mediate nitrogen catabolite repression in neurospora. mutants of nit ...19826125872
guanine uptake and metabolism in neurospora crassa.guanine is transported into germinated conidia of neurospora crassa by the general purine base transport system. guanine uptake is inhibited by adenine and hypoxanthine but not xanthine. guanine phosphoribosyltransferase (gprtase) activity was demonstrated in cell extracts of wild-type germinated conidia. the km for guanine ranged from 29 to 69 micro m in gprtase assays; the ki for hypoxanthine was between 50 and 75 micro m. the kinetics of guanine transport differ considerably from the kinetics ...19826174500
a simplified method for the simultaneous detection of intragenic and intergenic mutations (deletions) in neurospora crassa. 19826211615
circadian rhythms in neurospora crassa: a mutation affecting temperature compensation.the circadian rhythm of conidiation (spore formation) in neurospora crassa is known to be temperature compensated, that is, the period is only slightly affected by the incubation temperature. thus, the q10 (the relative rate enhancement corresponding to a 10 degrees c rise in temperature) of the rhythm of the bd csp strain from 14 to 30 degrees c was 1.1, whereas the q10 of the uncompensated growth rate in the same interval was 2.4. a mutation at the cel locus resulted in loss of the temperature ...19826461008
properties of mitochondria as a function of the growth stages of neurospora crassa.the oxidative and phosphorylative properties of mitochondria isolated from neurospora crassa were investigated as a function of growth stage. the rates of oxidation of exogenous nadh and nadph varied independently of each other, thus ruling out the existence of only one unspecific dehydrogenase. two different pathways were involved in the oxidation of nad-linked substrates, as indicated by changes in the rate of oxygen uptake, the sensitivity to rotenone, and the efficiency of phosphorylation. o ...19826460022
metabolic sequestration of putrescine in neurospora crassa. 19826462139
effects of divalent metal ions and chelators on the structure of outer mitochondrial membranes from neurospora crassa. 198219431484
genetic map of mitochondrial dna in podospora order to develop an eukaryotic vector with the podospora plasmid, further characterization is required of the mitochondrial dna into which this plasmid is integrated, a physical map (restriction sites) of the podospora chondriome (size 95 kb) has been completed. as prerequisite for the establishment of a genetic (functional) map, 70% of the chondriome was cloned in e. coli vectors. using mitochondrial genes from saccharomyces cerevisiae, six structural genes were located on the podospora chon ...198224186230
lipid and cell wall changes in an inositol-requiring mutant of neurospora inositol deficiency in the inositol-requiring (inl) mutant of neurospora crassa led to changes in the composition of the inositol-containing lipids and the cell wall. on deficient levels of inositol, phosphatidyl inositol decreased by 23-fold, di(inositolphosphoryl) ceramide decreased by 4-fold, and monoinositolphosphoryl ceramide increased slightly. the inositol deficiency also led to an aberrant hyphal morphology and changes in both the amount of cell wall and the amino sugar content of the ...1982220215
intron within the large rrna gene of n. crassa mitochondria: a long open reading frame and a consensus sequence possibly important in splicing.we describe the sequence of the 2295 nucleotide long intron and 245 nucleotides of the flanking exon sequences within the large (24s) rrna gene of neurospora crassa mitochondria. the intron contains a long open reading frame, which could correspond to ribosomal protein s5. comparison with the corresponding intron of the large rrna gene of yeast mitochondria reveals a single highly homologous 57 nucleotide long sequence, including the sequence (formula; see text), which is present in virtually al ...19826218884
extracellular acid proteases from neurospora crassa.three electrophoretically distinct acid proteases appear in culture filtrates of neurospora crassa. like the previously investigated alkaline and neutral proteases, these enzymes require induction by an exogenous protein. but in contrast to alkaline and neutral proteases, which are synthesized and secreted in response to limitation of any one of three nutrilites (carbon, nitrogen or sulfur), extracellular elaboration of the acidic proteases is more specifically a function of the missing nutrilit ...19826210687
primary structure of tyrosinase from neurospora crassa. i. purification and amino acid sequence of the cyanogen bromide fragments.cyanogen bromide (cb) cleavage of neurospora tyrosinase resulted in four major fragments, cb1 (222 residues), cb2 (82 residues), cb3 (68 residues), and cb4 (35 residues), and one minor overlap peptide cb2-4 (117 residues) due to incomplete cleavage of a methionylthreonyl bond. the sum of the amino acid residues of the four major fragments matches the total number of amino acid residues of the native protein. the amino acid sequences of the cyanogen bromide fragments cb2, cb3, and cb4 were determ ...19826210695
5'-untranslated sequences of two structural genes in the qa gene cluster of neurospora crassa.the coding regions of two genes (qa-2 and qa-3) in the qa gene cluster of neurospora crassa have been localized by nucleotide sequence analysis combined with data on previously determined nh2-terminal amino acid sequences for the proteins that these genes encode. the start point of transcription for each of these genes has been determined by nuclease s1 mapping experiments with poly(a)+rna isolated from quinic acid-induced cultures of n. crassa. the sequences of approximately 200 nucleotides 5' ...19826210913
the role of pyrimidine dimers in postreplication repair in neurospora.using the micrococcus luteus dimer specific endonuclease assay of wilkins (1973), and photoreactivation we have examined the induction and fate of ultraviolet induced pyrimidine dimers in the excision defective strain, uvs-2, of neurospora crassa. dimer induction was fluence dependent from 0 to 800 ergs/mm2 uv. an interdimer distance of 19.6 x 10(6) dna molecular weight was found after a fluence of 220 ergs/mm2. we confirm the earlier report that this mutant is completely excision defective (wor ...19826213836
comparison of the induction of specific locus mutations in wild-type and repair-deficient strains of neurospora crassa.a comparison of mutation induction between wild-type and excision repair-deficient strains has shown that, after treatment with four of the five mutagens tested, an enhanced recovery of induced mutants was found in the excision repair-deficient strains. in this sense we have confirmed for neurospora ames' (1977) observations with salmonella. furthermore, genetic analysis of the mutants induced in neurospora in both wild-type and excision repair-deficient strains has shown that in some cases the ...19826214249
characterization of neurospora crassa catabolic dehydroquinase purified from n. crassa and escherichia coli.1. neurospora crassa catabolic dehydroquinase has been purified from n. crassa and escherichia coli. 2. protein-sequence and gel-electrophoretic data show that apparently pure, homogeneous native dehydroquinase is a mixture of intact and proteinase-cleaved enzyme monomers. 3. protein-sequence data and steady-state kinetics show that the catabolic dehydroquinase gene of n. crassa is expressed with fidelity in e. coli.19826214255
an immunological study of the interaction of ligands with pyruvate kinase of neurospora crassa.antibodies against pyruvate kinase of neurospora crassa, induced in rabbits, were used to monitor the interaction of ligands with this enzyme. the technique of microcomplement fixation was employed to probe for conformational alterations elicited by binding of substrates (phosphoenolpyruvate (pep) and adenosine diphosphate), the allosteric activator (fructose 1,6-diphosphate), and the inhibitor (valine). on binding of pep and valine to pyruvate kinase a pronounced reduction in the extent of comp ...19826215112
the nucleotide sequence at the 3'-end of neurospora crassa 18s-rrna and studies on the interaction with 5s-rrna.the sequence of more than 100 nucleotides at the 3'-end of neurospora crassa 18s-rrna was determined by chemical sequencing techniques. extensive homologies with 18s-rrna from other eukaryotes were found. inspection of the nucleotide sequence at the 3'-end of n. crassa 5s-rrna revealed the presence of sequences complementary to a region near the 3'-terminus of 18s-rrna. under the appropriate conditions a complex was formed between 18s-rrna and 5s-rrna (tm 53 degrees c). interaction was detected ...19826217449
cell wall degradation in the autolysis of filamentous fungi.a systematic study on autolysis of the cell walls of fungi has been made on neurospora crassa, botrytis cinerea, polystictus versicolor, aspergillus nidulans, schizophyllum commune, aspergillus niger, and mucor mucedo. during autolysis each fungus produces the necessary lytic enzymes for its autodegradation. from autolyzed cultures of each fungus enzymatic precipitates were obtained. the degree of lysis of the cell walls, obtained from non-autolyzed mycelia, was studied by incubating these cell ...19826219290
isolation and properties of a cyclic amp-binding protein from neurospora. evidence for its role as the regulatory subunit of cyclic amp-dependent protein kinase.a cyclic amp-binding protein with a native molecular weight calculated to be 82,000 was purified 2,000-fold from neurospora crassa. the apparent subunit molecular weight was 47,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting that the native protein exists as a dimer of identically sized subunits. the 8-n3-cyclic [32p]amp-labeled protein appeared as a doublet upon two-dimensional gel electrophoresis, with pi = 5.4 and 5.5. binding studies with both noncyclic and cycli ...19826277955
cytochrome oxidase subunit iii gene in neurospora crassa mitochondria. location and sequence.we have located and sequenced the gene for cytochrome oxidase subunit iii (coiii) in neurospora crassa mitochondria. the coiii gene is located downstream from the small rrna gene within a cluster of trna genes and is coded by the same strand as the trna and the rrna genes. like the trna and the rrna genes, the coiii gene is also flanked by the gc-rich palindromic dna sequences which are highly conserved in n. crassa mitochondria. the coiii coding sequence predicts a protein 269 amino acids long ...19826279664
identification and isolation of actin from neurospora crassa.crude cell extracts of neurospora crassa contained an abundant protein that was identified as actin by a number of criteria. the protein, either in cell extracts or in pure form, co-migrated with rabbit skeletal muscle actin in polyacrylamide gels. the n. crassa actin was purified by deae-cellulose and dnaase i-sepharose chromatography and had the expected property of inhibiting dnaase i activity. although n. crassa actin could polymerize and depolymerize, purification based entirely on this cha ...19826281363
similar genes for a mitochondrial atpase subunit in the nuclear and mitochondrial genomes of neurospora crassa. 19826283377
1h nmr studies of eukaryotic cytochrome c. resonance assignments and iron-hexacyanide-mediated electron exchange.1h nmr resonance assignments in the spectra of horse, tuna, neurospora crassa and candida krusei cytochromes c are described. assignments have been made using nmr double-resonance techniques in conjunction with electron-exchange experiments, spectral comparison of related proteins, and consideration of the x-ray structure of tuna cytochrome c. resonances arising from 11 residues of horse cytochrome c have been assigned.19826284503
biogenesis of mitochondrial ubiquinol:cytochrome c reductase (cytochrome bc1 complex). precursor proteins and their transfer into mitochondria.the precursor proteins to the subunits of ubiquinol:cytochrome c reductase (cytochrome bc1 complex) of neurospora crassa were synthesized in a reticulocyte lysate. these precursors were immunoprecipitated with antibodies prepared against the individual subunits and compared to the mature subunits immunoprecipitated or isolated from mitochondria. most subunits were synthesized as precursors with larger apparent molecular weights (subunits i, 51,500 versus 50,000; subunit ii, 47,500 versus 45,000; ...19826286652
structural role of the tyrosine residues of cytochrome c.the tertiary structures of horse, tuna, neurospora crassa, horse [hse65,leu67]- and horse [hse65,leu74]-cytochromes c were studied with high-resolution 1h n.m.r. spectroscopy. the amino acid sequences of these proteins differ at position 46, which is occupied by phenylalanine in the horse proteins but by tyrosine in the remaining two, and at positions 67, 74 and 97, which are all occupied by tyrosine residues in horse and tuna cytochrome c but in the other proteins are substituted by phenylalani ...19826289807
cloning of the trp-1 gene from neurospora crassa by complementation of a trpc mutation in escherichia coli.studies with a hybrid plasmid containing 4.0 kilobase pairs of neurospora crassa dna cloned into plasmid pbr322 indicated that the plasmid restored to prototrophy a trpc mutant of escherichia coli which lacked phosphoribosyl anthranilate isomerase but not a trpc mutant which lacked indole glycerol phosphate synthetase, that the relevant transcription was initiated at a promoter within the n. crassa dna, and that the phosphoribosyl anthranilate isomerase could be specified by a subcloned segment ...19826290461
the mitochondrially made subunit 2 of neurospora crassa cytochrome aa3 is synthesized as a precursor protein. 19826291999
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
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
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
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
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
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 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 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
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
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
isolation of complex iii from various mitochondria. comparison of the structural and functional properties of the preparations from beef heart, calf liver and neurospora complex iii was isolated by an improved procedure from beef heart mitochondria, from neurospora crassa mitochondria and for the first time from mitochondria originating from mammalian tissue other than heart, i.e. calf liver. the described procedure consists of differential extraction of the respective mitochondria, hydroxyapatite chromatography and, finally, either gel- or affinity chromatography. the preparations contain the well known prosthetic groups, i.e. 6-8 mumol b-type heme, 3-4 ...19836321316
survey, purification, and properties of sugar phosphate phosphohydrolase among microorganisms.sugar phosphate phosphohydrolase was purified approximately 500- to 600-fold to apparent homogeneity from escherichia coli b, escherichia coli c, escherichia coli var. communior, escherichia acidilactici, enterobacter aerogenes, neisseria meningitidis, and saccharomyces cereviseae. the molecular weights of the enzyme as estimated by gel filtration ranged from 97 x 10(3) to 101 x 10(3). the enzyme was composed of two subunits with the same molecular weight which ranged from 50 x 10(3) to 52 x 10( ...19836322944
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
nucleotide sequence and intron structure of the apocytochrome b gene of neurospora crassa mitochondria.the sequence of the apocytochrome b (cob) gene of neurospora crassa has been determined. the structural gene is interrupted by two intervening sequences of approximately 1260 bp each. the polypeptide encoded by the exons shows extensive homology with the cob proteins of aspergillus nidulans and saccharomyces cerevisiae (79% and 60%, respectively). the two introns are, however, located at sites different from those of introns in the cob genes of a. nidulans and s. cerevisiae (which contain highly ...198310872314
a family of repetitive palindromic sequences found in neurospora mitochondrial dna is also found in a mitochondrial plasmid dna.neurospora mtdna contains a repetitive, 18 nucleotide palindromic sequence (5'-ccctgcagtactgcaggg-3') that contains two closely spaced psti sites (ctgcag) in the arms of the palindrome (yin, s., heckman, j., and rajbhandary, u. l. (1981) cell 26, 325-332). in the present study, dna sequence analysis was carried out to determine whether psti palindromes are present in an apparently distinct genetic element, the 3.6-kilobase mitochondrial plasmid from neurospora crassa strain mauriceville-1c (fgsc ...19836300080
structural variations and optional introns in the mitochondrial dnas of neurospora strains isolated from nature.mitochondrial dnas from ten wild-type neurospora crassa, neurospora intermedia, and neurospora sitophila strains collected from different geographical areas were screened for structural variations by restriction enzyme analysis. the different mtdnas show much greater structural diversity, both within and among species, than had been apparent from previous studies of mtdna from laboratory n. crassa strains. the mtdnas range in size from 60 to 73 kb, and both the smallest and largest mtdnas are fo ...19836300945
release of high molecular weight dna from neurospora crassa using enzymic digestions.methods are described that allow extraction of high molecular weight dna from germinated conidia of neurospora crassa. by labelling dna with ribonucleosides, early conidia were shown to be active in dna synthesis. these cells when treated with the enzyme zymolyase became fragile and could be readily lysed with ionic detergents to release high molecular weight dna. the dna extracted from zymolyase treated cells on to alkaline sucrose gradients sedimented as a heterogeneous species of up to 150 x ...19836302203
chimeric plasmid that replicates autonomously in both escherichia coli and neurospora crassa.a hybrid pbr322 plasmid (designated pdv1001) containing two functional escherichia coli antibiotic resistance genes (kanr and camr) and a qa-2+ gene from neurospora crassa transforms n. crassa qa-2- mutants to qa-2+ with a frequency of ca. 5 x 10(-5) per regenerated spheroplast (ca. 100 transformants per microgram of plasmid dna). this plasmid can replicate autonomously without integrating into the n. crassa genome. the autonomously replicating hybrid plasmid was detected in n. crassa transforma ...19836302666
construction of a shuttle vector for the filamentous fungus neurospora crassa.we have constructed a recombinant plasmid, pals-1, that replicates autonomously in both neurospora and escherichia coli. pals-1 consists of the mitochondrial plasmid from neurospora strain p405-labelle, the neurospora qa-2+ gene, and e. coli plasmid pbr325. pals-1 transforms the neurospora qa-2+ gene at frequencies 5- to 10-fold higher than those for plasmids that transform mainly by integration. when e. coli was transformed with dna from neurospora transformants, we recovered not only pals-1 bu ...19836302667
molybdenum cofactor in chlorate-resistant and nitrate reductase-deficient insertion mutants of escherichia coli.we examined molybdenum cofactor activity in chlorate-resistant (chl) and nitrate reductase-deficient (nar) insertion mutants and wild-type strains of escherichia coli k-12. the bacterial molybdenum cofactor was assayed by its ability to restore activity to the cofactor-deficient nitrate reductase found in the nit-1 strain of neurospora crassa. in the wild-type e. coli strains, molybdenum cofactor was synthesized constitutively and found in both cytoplasmic and membrane fractions. cofactor was fo ...19836307982
cloning of the structural gene for orotidine 5'-phosphate carboxylase of neurospora crassa by expression in escherichia coli.a neurospora gene bank in plasmid prk9 was used to complement pyrimidine auxotrophs in e. coli. two plasmids were obtained that complement a pyrf mutant of e. coli. these plasmids hybridise to neurospora dna and transform a pyr-4 strain of neurospora. the promoter used in expressing the orotidine 5'-monophosphate carboxylase in e. coli is within the neurospora sequence.19836308396
complete nucleotide sequence of the escherichia coli gdha gene.the dna sequence of the gdha gene of escherichia coli k12, which encodes the 447 amino acid polypeptide subunit of nadp-specific glutamate dehydrogenase, is presented. the deduced protein sequence is strongly homologous to the corresponding enzyme of the eukaryotic fungus neurospora crassa. the upstream dna sequence includes several overlapping promoter consensus sequences. the downstream dna sequence contains inverted repeats, predicted as forming long stable stem-loop structures in rna, homolo ...19836308576
receptor sites involved in posttranslational transport of apocytochrome c into mitochondria: specificity, affinity, and number of sites.assembly of cytochrome c involves a series of steps: synthesis of apocytochrome c on free ribosomes, specific binding of apocytochrome c to the mitochondrial surface, transfer across the outer membrane, covalent addition of protoheme, refolding of the polypeptide chain, and association of holocytochrome c with its functional sites at the inner membrane. the binding step of apocytochrome c to neurospora crassa mitochondria was studied by inhibiting the subsequent transfer steps with the heme anal ...19836308663
the genes coding for histone h3 and h4 in neurospora crassa are unique and contain intervening sequences.sequences coding for histone h3 and h4 of neurospora crassa could be identified in genomic digests with the use of the corresponding genes from sea urchin and x. laevis as hybridization probes. a 2.6 kb hindiii-generated n. crassa dna fragment, showing homology with the heterologous histone h3-gene probes was cloned in a charon 21a vector. using dna from this clone as a homologous hybridization probe a 6.9 kb sali-generated dna fragment was isolated which in addition to the histone h3-gene also ...19836310494
purification and properties of single strand dna-binding endo-exonuclease of neurospora crassa.single strand dna-binding endo-exonucleases purified from mitochondria, vacuoles, or a mixture of these organelles had the same high specific single strand dnase activity (910 mumol of nucleotides/min/mg), and each contained a polypeptide of mr = 31,000-33,000 which was found to be active by sodium dodecyl sulfate-dna-gel electrophoresis. the properties of the three preparations were identical in all respects tested. the enzyme showed distributive endonuclease activity with single strand dna, bu ...19836311833
cytochrome oxidase subunit 2 gene in neurospora crassa mitochondria.the nucleotide sequence of the cytochrome oxidase subunit 2 (cox2) gene has been obtained from cloned mitochondrial dna segments of neurospora crassa. the coding sequences have been identified on the basis of protein sequence homology with the subunit 2 of cytochrome oxidase from yeast and man. the postulated precursor of the n. crassa subunit 2 protein is 250 amino acids long, with a molecular weight of 28,700. as in the trna and rrna genes, the subunit 2 gene is flanked by g + c-rich palindrom ...19836313689
isolation of dna from filamentous fungi and separation into nuclear, mitochondrial, ribosomal, and plasmid components.a general procedure for purifying and efficiently separating four types of dna from filamentous fungi has been developed. the protocol involves (i) disruption of mycelial cells by blending in liquid nitrogen followed by suspension of cell contents in buffer containing high concentrations of protease and edta; (ii) deproteinization with phenol; (iii) cesium chloride/bisbenzimide density gradient centrifugation to separate nuclear dna, mitochondrial dna, and ribosomal dna; and (iv) agarose gel ele ...19836318603
transformation of aspergillus nidulans by the orotidine-5'-phosphate decarboxylase gene of neurospora crassa.relief of an auxotrophic requirement for uridine in aspergillus nidulans strain g191 has been achieved by transformation with a segment of neurospora crassa dna containing the corresponding gene coding for orotidine-5'-phosphate decarboxylase. the mitotic stability of such transformants suggests that the dna has integrated into the genome. southern hybridisation analysis of dna isolated from transformants revealed the presence of pbr322 sequences which have integrated into the host genome along ...19836220717
structure of the trifunctional trp-1 gene from neurospora crassa and its aberrant expression in escherichia coli.the trifunctional trp-1 gene from neurospora crassa was cloned by complementation of a phosphoribosylanthranilate isomerase-deficient mutant of e. coli. a 2.7-kb dna sequence containing trp-1 was determined. homology of the deduced trp-1 polypeptide sequence to the corresponding e. coli proteins is striking; the order of functional domains within trp-1 is nh2-glutamine amidotransferase-indoleglycerolphosphate synthase-phosphoribosylanthranilate isomerase-cooh (nh2-trpg-trpc-trpf-cooh). whereas t ...19836221060
control of the ornithine cycle in neurospora crassa by the mitochondrial neurospora crassa, the mitochondrial membrane separates ornithine used in arginine biosynthesis from ornithine used in the arginine degradative pathway in the cytosol. ornithine easily exchanges across the mitochondrial membrane under conditions appropriate for synthesis of the immediate biosynthetic product, citrulline. neither of the two mitochondrial enzymes required for the ornithine-to-citrulline conversion is feedback inhibitable in vitro. nevertheless, when arginine is added to cells a ...19836222031
ribosomal rna genes of neurospora crassa: multiple copies and specificities.ribosomal rna genes were isolated from the germinated conidial and mycelial cells of n. crassa by repeated cycles of 3h-dna:rrna reactions followed by hydroxyapatite chromatography. specificity of multiple copies of those rdnas with respect to n. crassa cell types was studied. the fraction of n. crassa germinated conidial in vitro labelled 3h-dna recovered in the presence of rrna isolated from the same cell type was about 2.2%, when compared with approximately 1.2% rdnas obtained in mycelial cel ...19836222241
[microbiological short-time tests for the evaluation of mutagenic potential of chemical substances].during the last 20 years it became much more interesting to test new chemicals as fast as possible for their carcinogenic potency. therefore new test models were developed. mutagenicity seems to be one sign for carcinogenicity. therefore test systems using microorganisms were studied which are influenced by mutagenic substances. these systems are described, first of all the ames-test, using revertants of salmonella typhimurium, secondly the escherichia coli system deficient of dna-polymerase a ( ...19836222262
purification and characterization of an extracellular acid protease from neurospora extracellular acid protease was purified 1420-fold from sulfur-starved protein-induced cultures of neurospora crassa. the enzyme was homogeneous as determined by polyacrylamide electrophoresis. the purification procedure consisted of an ultrafiltration step, cation-exchange chromatography, and affinity chromatography on sepharose-linked pepstatin. the enzyme is homologous to aspartyl proteases that are characterized by pepstatin inhibition and trypsinogen activation. it is extremely autolytic ...19836222698
evolutionary aspects of accuracy of phenylalanyl-trna synthetase. a comparative study with enzymes from escherichia coli, saccharomyces cerevisiae, neurospora crassa, and turkey liver using phenylalanine analogues.the phenylalanyl-trna synthetases from escherichia coli, saccharomyces cerevisiae, neurospora crassa, and turkey liver activate a number of phenylalanine analogues (tyrosine, leucine, methionine, p-fluorophenylalanine, beta-phenylserine, beta-thien-2-ylalanine, 2-amino-4-methylhex-4-enoic acid, mimosine, n-benzyl-l- or n-benzyl-d-phenylalanine, and ochratoxin a), as demonstrated by km and kcat of the atp/ppi pyrophosphate exchange. upon complexation with trna, the enzyme-trnaphe complexes show a ...19836222761
a study of the heat-shock response in neurospora crassa.1. neurospora crassa was grown at 28 degrees c for 12 hr and transferred to higher temperatures for 2 hr. 2. cultures labelled with [35s]methionine showed the synthesis of several new proteins in response to heat-shock at 46 to 48 degrees c. 3. major polypeptides of approximate mr 105,000, 99,000, 78,000, 43,000 and 23,000 were detectable in one-dimensional sds-polyacrylamide slab gel electropherograms. 4. 2-d analysis using isoelectric-focussing in the first dimension and electrophoresis in sds ...19836222926
[isolation and characteristics of the nucleosomes from the mold neurospora crassa]. 19836227467
biosynthesis and assembly of nuclear-coded mitochondrial membrane proteins in neurospora crassa. 19836228709
a colony filter-hybridization procedure for the filamentous fungus neurospora crassa.a colony filter-hybridization procedure for the filamentous fungus neurospora crassa has been developed. the procedure is sensitive enough to detect escherichia coli plasmid pbr322 dna integrated into chromosomal dna in a neurospora transformant. thus, it should facilitate the isolation of nuclear genes by plasmid-rescue procedures.19836229196
radioassay of the folate-hydrolyzing enzyme activity, and the distribution of the enzyme in biological cells and tissues.a sensitive radioassay method has been developed to quantitate the activity of the folate-hydrolyzing enzyme which catalyzes the hydrolysis of folic acid to pteroic acid and glutamic acid. the method is based on analyzing [2-14c]pteroic acid separated by a thin-layer chromatography on an avicel sf cellulose plate using 0.1 m potassium phosphate buffer, ph 7.0, as a solvent. this method was found to be more sensitive than a conventional photometric method to determine the activity of the folate-h ...19836229614
molecular dosimetry of the chemical mutagen ethyl methanesulfonate. quantitative comparison of the mutagenic potency in neurospora crassa and saccharomyces cerevisiae.extending previous work with e. coli and mammalian cells in culture, forward-mutation frequencies induced by ethyl methanesulfonate (ems) were quantitatively compared in neurospora crassa and saccharomyces cerevisiae under standardized conditions. concomitantly, the actual dose to dna was measured by determining the amount of radioactivity bound to dna after treatment with tritium-labeled ems. after exposure to ems (2.5-50 mm), alkylation levels in n. crassa and s. cerevisiae were similar to tho ...19836218404
peptide utilization by nitrogen-starved neurospora crassa.peptides ranging in size from a mean number of 30 residues down to dipeptides supported growth of a leucine auxotroph when used as both a nitrogen and leucine source. under nitrogen-limiting conditions, the peptides induced extracellular peptidohydrolytic activity, hydrolyzing peptides to monomer amino acids. growth of a leu-2 mutant of neurospora crassa on those peptides transportable by the oligopeptide transport system did not result in induction of hydrolytic activity, whereas growth of a le ...19836219099
the complete nucleotide sequence of the neurospora crassa am (nadp-specific glutamate dehydrogenase) gene.the complete nucleotide sequence of a 2.7-kb genomic fragment, containing the neurospora crassa am [nadp-specific glutamate dehydrogenase (gdh)] gene, has been determined. the transcription initiation and polyadenylation sites have been defined by s1 mapping. there are at least four initiation sites between 35 and 60 bases downstream of a tataaa sequence. the single polyadenylation site is immediately downstream of a six-nucleotide sequence which is present in the corresponding position in the n ...19836231215
mutagenesis at the ad-3a and ad-3b loci in haploid uv-sensitive strains of neurospora crassa. vi. genetic characterization of ad-3 mutants provides evidence for qualitative differences in the spectrum of genetic alterations between wild-type and nucleotide excision-repair-deficient strains.genetic characterization of ad-3b mutants induced in wild-type and uv-sensitive strains has revealed qualitative differences between the spectra of genetic alterations at the molecular level. ad-3b mutants induced in the two nucleotide excision-repair-deficient strains upr-1 and uvs-2 (worthy and epler, 1973) had significantly lower frequencies of nonpolarized complementation patterns and higher frequencies of noncomplementing mutants than ad-3b mutants induced in the wild-type strain in samples ...19836188039
a chicken repetitive dna sequence that is highly sensitive to single-strand specific endonucleases.a dna sequence consisting of the 5-mer agagg repeated tandemly 32 times has been detected in a chicken genomic clone and found to be present in about 2000 copies per chicken genome. this sequence was highly susceptible to single-strand specific endonucleases isolated from aspergillus oryzae (s1) and mung bean, but cleavage by a single-strand specific endonuclease isolated from neurospora crassa occurred only at a ph below 5.5. endonucleolytic cutting of the agagg sequence by the single-strand sp ...19836231528
calcium inhibition of a heat-stable cyclic nucleotide phosphodiesterase from neurospora crassa.neurospora crassa had a heat-stable (up to 95 degrees c), soluble cyclic nucleotide phosphodiesterase (pde). both unheated and heat-stable pde activities were inhibited by micromolar concentrations of ca2+. this inhibition was reversed by egta or edta in molar excess of the ca2+ concentration. calmodulin was not involved in the ca2+ inhibition, nor was ca2+ inhibition of the heat-stable pde due to cleavage inactivation of the enzyme by a ca2+-stimulated protease. in addition to ca2+, several oth ...19836298002
comparison of the vacuolar membrane atpase of neurospora crassa with the mitochondrial and plasma membrane atpases.the vacuolar membrane atpase of neurospora crassa closely resembles the mitochondrial atpase in its substrate specificity, substrate affinity, and sensitivity to the inhibitor n,n'-dicyclohexylcarbodiimide. three different mutants with altered mitochondrial atpase activity, exhibited as 1) resistance to n,n'-dicyclohexylcarbodiimide, 2) enhanced sensitivity to n,n'-dicyclohexylcarbodiimide, and 3) very low specific activity, were found to be unaltered in the vacuolar membrane atpase. the vacuola ...19836228553
possible role of a regulatory gene product upon the myo-inositol-1-phosphate synthase production in neurospora crassa.the regulatory effect of inositol on inositol-1-phosphate synthase in neurospora crassa strains was studied. inositol represses enzyme production in the cultures of the wild type and that of the thermosensitive inositol-requiring mutant grown at 22 degrees c. enzyme activity as well as the quantity of enzyme protein decreased sharply in both strains by increasing concentrations of inositol in the medium. inositol-requiring strains used in our experiments can be divided into two groups. the first ...19836228255
purification and properties of a low molecular weight dna polymerase from neurospora crassa.a third dna polymerase 'c' with low molecular weight was isolated and purified 3700-fold from ground hyphae of neurospora crassa wt 74 a, which shows similarities to beta- and gamma-polymerases from higher eukaryotes: preference for poly(ra)(dt) as a template/primer, inhibition by p-chloromercuribenzoate, resistance against n-ethylmaleimide up to 10 mmol/l, and molecular weight of about 40000. this polymerase elutes as a distinct peak from deae-cellulose at 0.60 mol/l kcl and has an optimum for ...19836197088
point mutations and dna rearrangements 5' to the inducible qa-2 gene of neurospora allow activator protein-independent transcription.expression of the qa-2 gene of neurospora crassa normally requires a functional activator protein encoded by qa-1f. twelve transcriptional mutants of the qa-2 gene have been isolated in qa-1f- strains, and these allow partial expression of qa-2 (1-45% of induced wild type) in the absence of functional activator protein. all 12 mutants have been characterized by genomic (southern) blot hybridization and the dnas of 5 have been cloned and sequenced. eight mutations consist of large dna rearrangeme ...19836316356
genetic and biochemical characterization of glutamine synthetase from neurospora crassa glutamine auxotrophs and their this paper we present the isolation and characterization of glutamine auxotrophs of neurospora crassa and their revertants. the results show that although various enrichment procedures were used, we found only two types of auxotrophs. genetic crosses performed between the different mutants showed that the mutations responsible for their phenotypes were highly linked and probably affected the same gene. the biochemical characterization of the glutamine synthetase polypeptides of the different ...19836139363
isolation and characterization of plasma membranes from strains of neurospora crassa with wild type morphology.a variety of commercially available cell wall hydrolytic enzyme preparations were screened alone and in various combinations for their ability to degrade the cell wall of neurospora crassa wild type strain 1a. a combination was found which causes complete conversion of the normally filamentous germinated conidia to spherical structures in about 1.5 h. examination of these spheroplasts by scanning electron microscopy indicated that, although they are spherical, they retain a smooth coat that can ...19836227620
precursor proteins are transported into mitochondria in the absence of proteolytic cleavage of the additional sequences.many nuclear-coded mitochondrial proteins are synthesized as larger precursor polypeptides that are proteolytically processed during import into the mitochondrion. this processing appears to be catalyzed by a soluble, metal-dependent protease localized in the mitochondrial matrix. in this report we employ an in vitro system to investigate the role of processing in protein import. intact neurospora crassa mitochondria were incubated with radiolabeled precursors in the presence of the chelator o-p ...19836226666
kinetic evidence for interacting active sites in the neurospora crassa plasma membrane atpase.the rate of mgatp hydrolysis (v) by the neurospora plasma membrane atpase shows a sigmoid relationship to substrate concentration ( [s] ), which is precisely fit by the equation: v = (vmax x [s]2)/(km + [s]2). this equation describes an enzyme with two substrate-binding sites, both of which must be filled for hydrolysis to occur. at concentrations above 1 mm, both free mg2+ and free atp behave as competitive inhibitors of the atpase. free atp, although not hydrolyzed, can also significantly stim ...19836226663
temperature-induced modifications of glycosphingolipids in plasma membranes of neurospora crassa.plasma membranes isolated from a cell-wall-less mutant of neurospora crassa grown at 37 and 15 degrees c display large differences in lipid compositions. a free sterol-to-phospholipid ratio of 0.8 was found in 37 degrees c membranes, while 15 degrees c plasma membranes exhibited a ratio of nearly 2.0. membranes formed under both growth conditions were found to contain glycosphingolipids. cultures grown at the low temperature, however, were found to contain 6-fold higher levels of glycosphingolip ...19836226315
transient kinetic studies of neurospora crassa cytochrome c oxidase.the reaction of neurospora crassa cytochrome c oxidase with co was studied by flash-photolysis and rapid-mixing experiments, leading to the determination of the association and dissociation rate constants (7 x 10(4) m-1 x s-1 and 0.02s-1 respectively). pre-steady-state kinetic investigations of the catalytic properties of the enzyme showed that under proper conditions neurospora cytochrome c oxidase can be 'pulsed', i.e. activated, like the mammalian enzyme. the 'pulsed' species is spectroscopic ...19836316928
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