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gtp is required for the integration of a fragment of the neurospora crassa h(+)-atpase into homologous microsomal vesicles.the integration of a fragment of the neurospora crassa plasma membrane h(+)-atpase was examined to determine if insertion of the fragment into homologous microsomal vesicles is obligatorily dependent on a nucleoside triphosphate. rna transcripts that encoded the amino terminal 344 amino acids of the neurospora crassa plasma membrane h(+)-atpase(pma(344)+) were translated in a n. crassa in vitro system. the pma(344)+ integrated post-translationally into homologous microsomal vesicles independent ...19911711898
the cdna sequence and expression of an ubiquitin-tail gene fusion in neurospora crassa.the genome of neurospora crassa contains at least one natural fusion gene encoding a single ubiquitin (ubi) unit with a 78-amino acid c-terminal extension. the predominantly basic tail sequence corresponds to a highly conserved ribosomal protein identified in other organisms. the 0.7-kb ubi fusion transcript is mainly expressed in germinating conidia and other stages of active cell replication. under starvation conditions attained by nutrient depletion, or after polyamine depletion, the ubi fusi ...19911650731
nit-3, the structural gene of nitrate reductase in neurospora crassa: nucleotide sequence and regulation of mrna synthesis and turnover.the nit-3 gene of the filamentous fungus neurospora crassa encodes the enzyme nitrate reductase, which catalyzes the first reductive step in the highly regulated nitrate assimilatory pathway. the nucleotide sequence of nit-3 was determined and translates to a protein of 982 amino acid residues with a molecular weight of approximately 108 kda. comparison of the deduced nit-3 protein sequence with the nitrate reductase protein sequences of other fungi and higher plants revealed that a significant ...19911829499
effects of heat shock on the level of trehalose and glycogen, and on the induction of thermotolerance in neurospora crassa.neurospora crassa conidiospore germlings exposed to a heat shock (30-45 c) rapidly accumulated trehalose and degraded glycogen, even in the presence of cycloheximide. this phenomenon was also rapidly reversible upon return of the cells at 30 degrees c. trehalose accumulation at 45 degrees c demanded an exogenous source of carbon and either glucose or glycerol fulfilled such requirement. experiments with the cyclic amp-deficient cr-1 mutant suggested that the effects of temperature shifts on treh ...19911645296
isolation of uridine auxotrophs from trichoderma reesei and efficient transformation with the cloned ura3 and ura5 genes.uridine auxotrophs of the filamentous fungus trichoderma reesei have been selected using a positive screening procedure with 5-fluoro orotate. mutants deficient for the orotidine-5'-phosphate decarboxylase gene (ura3 mutants) and for the orotate phosphoribosyl transferase gene (ura5 mutants) have been characterized. the homologous ura3 and ura5 genes have been isolated and used to transform the auxotrophic mutants. transformation efficiency with these homologous systems is very high (greater tha ...19911913875
regulation of laccase synthesis in induced neurospora crassa cultures.rapidly growing cultures of n. crassa do not produce laccase. exposure of this fungus to different inducing agents leads to a de novo biosynthesis of extracellular laccase in vegetative cultures. in this study the induction of laccase after addition of cycloheximide and d-phenylalanine is reported. de novo synthesis of laccase mrna was followed over 96 h after induction. a fast appearance of the message, as well as its presence over a rather long period, indicates a regulation on a transcription ...19911833078
determination of electric parameters of cell membranes by a dielectrophoresis method.marszalek, p., j. j. zielinsky, and m. fikus (1989. bioelectrochem. bioenerg. 22:289-298) have described a novel design for measuring the complete dielectrophoretic spectrum of a single cell. from the analysis of the dielectrophoretic spectrum, the membrane conductivity, sigma membr, and the membrane dielectric permittivity, epsilon membr, of the cell may be determined according to the theory of dielectrophoresis described by sauer, f. a. (1985. interactions between electromagnetic field and cel ...19911831052
photostimulation of conidiation in mutants of neurospora crassa.various mutants of neurospora crassa were screened for light-stimulated conidiation which is a blue light effect and, at least in strain albino-band, is mediated by the flavoprotein nitrate reductase (nr). nr- mutants showed practically no photoconidiation under standard conditions. however, in fusion products of nit-1 (diaphorase activity present, terminal activity missing) plus nit-3 (terminal activity present, diaphorase activity missing), nr activities and photoconidiation were partially res ...19911830899
loss of nad(p)-reducing power and glutathione disulfide excretion at the start of induction of aerial growth in neurospora crassa.when exponentially growing hyphae of neurospora crassa in aerated liquid cultures are filtered and the resulting mycelial mat is exposed to air, aerial hyphae develop and synchronous conidiation is obtained. the hyphae in direct contact with air adhere to each other within minutes and form aerial hyphae during the following 12 h; the hyphae which are not in direct contact with air do not adhere to each other and do not form aerial hyphae. previous data indicated that oxidative stress was generat ...19911827113
an inducible gene expression system for neurospora crassa.neurospora crassa acetyl coa synthetase is highly induced when the growing mycelium is transferred from sucrose- to acetate-based medium. the inducible promoter of this gene has been isolated and used to control the expression of glutamate dehydrogenase. transformants containing this expression cassette show gdh levels up to 25 times higher than the nontransformed host strain. this expression cassette will form the basis of a system of heterologous gene expression.19911369451
regulation of biosynthesis of l-amino acid oxidase by neurospora crassa.l-amino acid oxidase was purified from liquid cultures of neurospora crassa induced by l-phenylalanine, d-phenylalanine, atp and cycloheximide. although the four enzyme species isolated were found to differ in size and electrophoretic mobility, they were the product of a single gene as demonstrated by genomic southern analysis. northern analysis of total cellular rna showed a rapid increase of l-amino acid oxidase mrna in response to the different inducing agents studied. these data suggest that ...19911829425
metalloselenonein, the selenium analogue of metallothionein: synthesis and characterization of its complex with copper ions.we used an automated peptide synthesizer to produce a peptide, metalloselenonein, that contains selenocysteine residues substituted for all cysteine residues in neurospora crassa copper metallothionein. metalloselenonein binds 3 mol of cu(i) per mol. this adduct shows a broad absorption band between 230 and 400 nm and a fluorescence band at 395 nm, which can be attributed to copper-selenolate coordination. the circular dichroism spectrum of the copper-metalloselenonein complex shows a positive b ...19911826562
ca2+ calmodulin-dependent protein kinase activity in the ascomycetes neurospora crassa.deae-cellulose column chromatography of neurospora crassa soluble mycelial extracts leads to the resolution of three major protein kinase activity peaks designated pki, pkii, and pkiii. pkii activity is stimulated by ca2+ and neurospora or brain calmodulin. maximal stimulation was observed at 2 microm-free ca2+ and 1 microgram/ml of the modulator. the stimulatory effect of the ca(2+)-calmodulin complex was blocked by egta and by some calmodulin antagonists such as phenothiazine drugs or compound ...19911652680
regulation of molybdenum cofactor species in the green alga chlamydomonas reinhardtii.molybdenum cofactor (moco) of molybdoenzymes is constitutively produced in cells of the green alga chlamydomonas reinhardtii grown in ammonium media, under which conditions certain molybdoenzymes are not synthesized. in soluble form, moco was found to be present in several forms: (i) as a low mr free species; (ii) bound to a moco-carrier protein of about 50 kda that could release moco to directly reconstitute in vitro nitrate reductase activity in the nit-1 mutant of neurospora crassa, but not t ...19911826614
nucleotide and dna uptake by neurospora crassa: involvement of an uptake stimulating protein.the basal and dusf (dna-uptake-stimulating factor, described previously by schablik and szabó (1981) fems microbiol. lett. 10, 395-397) stimulated uptake of [3h]dna and radioactive nucleotides by neurospora crassa (fgsc 1118, slime) cell-wall-less strain was studied. the uptake of [3h]dna by the cells is a saturable and time-dependent process. the ph and temperature optimum for [3h]dna uptake are ph 7 and 27 degrees c, respectively. both basal and dusf-stimulated uptake of [3h]dna are inhibited ...19911826455
specificity of repeat-induced point mutation (rip) in neurospora: sensitivity of non-neurospora sequences, a natural diverged tandem duplication, and unique dna adjacent to a duplicated region.the process designated rip (repeat-induced point mutation) alters duplicated dna sequences in the sexual cycle of neurospora crassa. we tested whether non-neurospora sequences are susceptible to rip, explored the basis for the observed immunity to this process of a diverged tandem duplication that probably arose by a natural duplication followed by rip (the neurospora zeta-eta region), and investigated whether rip extends at all into unique sequences bordering a duplicated region. bacterial sequ ...19911827630
recurrence of repeat-induced point mutation (rip) in neurospora crassa.duplicate dna sequences in the genome of neurospora crassa can be detected and mutated in the sexual phase of the life cycle by a process termed rip (repeat-induced point mutation). rip occurs in the haploid nuclei of fertilized, premeiotic cells before fusion of the parental nuclei. both copies of duplications of gene-sized sequences are affected in the first generation at frequencies of approximately 50-100%. we investigated the extent to which sequences altered by rip remain susceptible to th ...19911827629
surface topography and molecular stoichiometry of the mitochondrial channel, vdac, in crystalline arrays.the mitochondrial outer membrane contains a protein, called vdac, that forms large aqueous pores. in neurospora crassa outer membranes, vdac forms two-dimensional crystalline arrays whose size and frequency can be greatly augmented by lipase treatment of these membranes (c. mannella, science 224, 165, 1984). fourier filtration and surface reconstruction of freeze-dried/shadowed (45 degrees) arrays produced detailed images of two populations of crystals, whose lattices are mirror images of each o ...19911725124
purification and characterization of glutamate decarboxylase from neurospora crassa conidia.l-glutamate decarboxylase, an enzyme under the control of the asexual developmental cycle of neurospora crassa, was purified to homogeneity from conidia. the purification procedure included ammonium sulfate fractionation and deae-sephadex and cellulose phosphate column chromatography. the final preparation gave a single band on sodium dodecyl sulfate-polyacrylamide gels with a molecular weight of 33,200 +/- 200. a single band coincident with enzyme activity was found on native 7.5% polyacrylamid ...19911825829
kinetic properties of de-repressible alkaline phosphatase (ec 3.1.3.1) from neurospora crassa.de-repressible alkaline phosphatase from n. crassa shows inhibition by pnp-p and a hyperbolic mixed-type inhibition by pi. both increasing concentrations of pi and decreases in assay ph abolished inhibition by the substrate. also, pi promoted polymerization of the enzyme molecule, whose effect may account for the inhibitory behaviour shown by the enzyme in the presence of low pi concentrations.199124424939
polyamine toxicity in neurospora crassa: protective role of the vacuole.we used mutant strains of neurospora crassa to define the discretionary capacity of this species for excess putrescine. the spe-3 mutant, which accumulates putrescine internally, and the puu-1 mutant, which accumulates toxic levels of putrescine from the medium, both sequestered large excesses of putrescine in vacuoles. concomitantly in puu-1, inorganic polyphosphate increased modestly and some of the monovalent cation of the vacuole was discharged. these two factors contribute to the increased ...19911832828
calcium modulation of polyamine transport is lost in a putrescine-sensitive mutant of neurospora crassa.putrescine transport in neurospora is saturable and concentrative in dilute buffers, but in the growth medium putrescine simply equilibrates across the cell membrane. we describe a mutant, puu-1, that can concentrate putrescine from the growth medium because the polyamine transport system has lost its normal sensitivity to ca2+. the wild type closely resembles the mutant if it is washed with citrate and ethylene glycol bis(beta-aminoethyl ether)n,n'-tetraacetic acid. the mutant phenotype also ap ...19911832827
purification of nad glycohydrolase from neurospora crassa conidia by a polyclonal immunoadsorbent.nad glycohydrolase from neurospora crassa conidia was purified by affinity chromatography on a column of polyclonal antibodies bound to an agarose matrix. the procedure was easy, non-denaturating and suitable for repetitive use of the gel. the enzyme obtained appeared homogeneous by sodiumdodecyl sulphate-polyacrylamide gel electrophoresis.19911828469
31p-nmr-studies on intracellular ph and metabolite concentrations in relation to the circadian rhythm, temperature and nutrition in neurospora crassa.perfused mycelia of neurospora crassa were analyzed in vivo with 31p-nmr. both the cytoplasmatic and the vacuolar ph and the concentrations of phosphate metabolites were followed up to 30 h under constant conditions. no circadian changes were detected. however, slight changes in the nutrition or oxygen supply induced distinct changes in the intracellular ph and in the concentrations of metabolites. an increase of temperature from 21 to 43 degrees c lowered the intracellular ph and the metabolite ...19911825790
nucleotide sequence, messenger rna stability, and dna recognition elements of cys-14, the structural gene for sulfate permease ii in neurospora crassa.the complete nucleotide sequence of the cys-14 gene which encodes sulfate permease ii, a member of the sulfur regulatory circuit, is presented. the cys-14 gene contains four introns with consensus splice site sequences and is transcribed from four closely spaced initiation sites located approximately 20 bp upstream of the atg initiation codon. the translated cys14 protein is composed of 781 amino acids with a molecular weight of 87,037 and contains 12 potential hydrophobic membrane-spanning doma ...19911825178
cdna and genomic dna sequence of the 21.3 kda subunit of nadh:ubiquinone reductase (complex i) from neurospora crassa.the primary structure of a nuclear-encoded subunit of the respiratory chain nadh:ubiquinone reductase (complex i) from neurospora crassa was determined by sequencing cdna, genomic dna and the n-terminus of the protein. the sequence correlates to a protein of 200 amino acids and a molecular mass of 21349 da. the protein is synthesized without a cleavable presequence. it contains two alpha-helices predicted to traverse the bilayer and is a constituent of the membrane part of complex i.19911825789
purification and properties of a casein kinase ii-like enzyme from neurospora crassa.a serine/threonine protein kinase was partially purified from neurospora crassa. its physical and catalytic properties were typical of casein kinase ii. in vitro, the kinase phosphorylated a calpain like protease from allomyces arbuscula with higher affinity than a mixture of caseins.19911827768
a new senescence-inducing mitochondrial linear plasmid in field-isolated neurospora crassa strains from india.several field-collected strains of neurospora crassa from the vicinity or aarey, bombay, india, are prone to precocious senescence and death. analysis of one strain, aarely-1e, demonstrated that the genetic determinants for the predisposition to senescence are maternally inherited. the senescence-prone strains contain a 7-kb, linear, mitochondrial dna plasmid, maranhar, which is not present in long-lived isolates from the same geographical location. the maranhar plasmid has inverted terminal rep ...19911648454
regulation of the proton/electron stoichiometry of mitochondrial ubiquinol:cytochrome c reductase by the membrane potential.the electron transfer reaction catalysed by mitochondrial ubiquinol:cytochrome c reductase is linked to the outwards translocation of protons with an h+ e- stoichiometry of 1 under non-membrane potential condition. the effect of the electrical membrane potential on the h+/e- stoichiometry was investigated. the enzyme was isolated from neurospora crassa, reconstituted into phospholipid vesicles and electrical membrane potentials of various values were generated across the membranes by means of th ...19911847681
purification and properties of a membrane-bound insulin binding protein, a putative receptor, from neurospora crassa.the protein that is responsible for specific, high-affinity binding of insulin to the surface of neurospora crassa cells has been purified to homogeneity. the insulin binding activity of solubilized plasma membranes resembled that of intact cells with regard to affinity of binding, specificity for mammalian insulins, and amount of insulin bound per cell. insulin binding activity was purified from triton x-100 solubilized membranes in two steps: fplc on a monoq hr5/5 column; and affinity chromato ...19911824821
relationship between a subunit of nadh dehydrogenase (complex i) and a protein family including subunits of cytochrome reductase and processing protease of mitochondria.the primary structure of a 40 kda subunit of the respiratory chain nadh:ubiquinone reductase from neurospora crassa was determined by sequencing cdna, genomic dna and the n-terminus of the mature protein. the gene which is interrupted by 7 introns encodes a preprotein consisting of 375 amino acids with a 26 amino acid long presequence typical for a mitochondrial targeting signal. the sequence of the mature subunit shows conspicuous similarities to the recently [(1989) nature 339, 147-149] discov ...19911825202
nuclease-induced dna structural changes assessed by flow cytometry with the intercalating dye propidium iodide.a flow cytometric analysis of dna structural changes induced by cleavage with nucleases was performed on isolated hela nuclei by assessing changes in stainability with the dna-specific fluorochrome propidium iodide (pi). after mild digestion with dnase i, micrococcal nuclease, or with the single-strand-specific s1 and neurospora crassa nucleases, fluorescence intensity of nuclei stained with pi increased by about 15-30% above the value of undigested control samples. no significant modifications ...19912065557
sequence of the cloned pyr4 gene of trichoderma reesei and its use as a homologous selectable marker for transformation.we have cloned and sequenced the trichoderma reesei pyr4 gene encoding orotidine-5'-monophosphate decarboxylase. comparison of this sequence with that of the equivalent gene from other filamentous fungi suggests that t. reesei is closely related to cephalosporium acremonium and neurospora crassa. the cloned pyr4 gene has been used as a homologous selectable marker for transformation of t. reesei. the majority of transformants obtained with circular plasmid were mitotically unstable and contained ...19912036683
effect of activators and inhibitors on the activity of mitochondrial dna polymerase.at a concentration of 0.5 to 3 mmol/l, atp stimulates the activity of mitochondrial dna polymerase of neurospora crassa under the optimum reaction conditions; at higher concentrations, an inhibitory effect is observed. 4-chloromercuribenzoate (1 mmol/l), a thiol inhibitor, decreases the enzyme activity two-fold, while n-ethylmalcimide (2 mmol/l) has no effect. ethidium bromide (up to 10 mumol/l) and heparin (up to 0.4 micrograms/ml) reduce the activity by 60%. ddttp does not affect the dna polym ...19911841856
purification of neurospora crassa copper-metallothionein. 19911838130
amplitude model for the effects of mutations and temperature on period and phase resetting of the neurospora circadian oscillator.this paper analyzes published and unpublished data on phase resetting of the circadian oscillator in the fungus neurospora crassa and demonstrates a correlation between period and resetting behavior in several mutants with altered periods: as the period increases, the apparent sensitivity to resetting by light and by cycloheximide decreases. sensitivity to resetting by temperature pulses may also decrease. we suggest that these mutations affect the amplitude of the oscillator and that a change i ...19911837742
temperature compensation of the circadian rhythm in neurospora crassa: comments on the vicinal water hypothesis. 19911837741
genetics and molecular biology of neurospora crassa. 19911837201
expression of the escherichia coli beta-glucuronidase gene in pseudocercosporella herpotrichoides.the plant-pathogenic fungus pseudocercosporella herpotrichoides has been successfully transformed by using two different positive selection systems in combination with the escherichia coli gusa gene. the selectable markers used in this study were the hygromycin b phosphotransferase gene (hph) from e. coli and the gene (bml) for beta-tubulin from a benomyl-resistant mutant of neurospora crassa. a lower transformation rate was obtained with the bml system than with the hph system. conversely, cotr ...19911746951
heterologous expression of the aspergillus nidulans regulatory gene nira in fusarium oxysporum.we have isolated strains of fusarium oxysporum carrying mutations conferring a phenotype characteristic of a loss of function in the regulatory gene of nitrate assimilation (nira in aspergillus nidulans, nit-4 in neurospora crassa). one of these nir- mutants was successfully transformed with a plasmid containing the nira gene of a. nidulans. the nitrate reductase of the transformants is still inducible, although the maximum activity is lower than in the wild type. single and multiple integration ...19911756977
cloning of a sequence of aquaspirillum magnetotacticum that complements the arod gene of escherichia coli.a 2 kb dna fragment isolated from a cosmid library of aquaspirillum magnetotacticum strain ms-1 complements the aromatic-metabolite requirements and iron-uptake deficiencies of escherichia coli and salmonella typhimurium strains that lack a functional arod (biosynthetic dehydrodquinase) sequence. all recombinant cosmids selected for their arod complementation property carry this sequence. no dna sequence homology has, however, been detected by southern hybridization between the cloned fragment a ...19911766390
[synthesis and antimicrobial activity of substituted fluorobenzyl benzylidenethiazolidinediones and imidazolidinediones].the synthesis of six benzylidene thiazolidine-diones and three benzylidene imidazolidine-diones is described. in order to investigate their antimicrobial activity, they are evaluated against micro-organism such as staphylococcus aureus, streptococcus feacalis, mycobacterium smegmatis and neurospora crassa.19911795213
x-ray-induced specific-locus mutations in the ad-3 region of two-component heterokaryons of neurospora crassa, ix. mutational spectra as a function of x-ray dose.in previous studies, x-ray-induced specific-locus mutations in the adenine-3 (ad-3) region of a two-component heterokaryon (h-12) of neurospora crassa were combined with a series of tester strains carrying markers in the ad-3 and immediately adjacent regions to map mutants that were presumed multilocus deletions (de serres, 1989c, 1990a). two new classes of x-ray-induced mutations were recovered: multiple-locus mutations consisting of gene/point mutations at the ad-3a or ad-3b locus with a close ...19911824717
mutagenic potency and specificity of procarbazine in the ad-3 forward-mutation test in growing cultures of heterokaryon 12 of neurospora crassa.procarbazine (natulan) was tested for its mutagenic potency and specificity in the ad-3 forward-mutation test in heterokaryon 12 (h-12) of neurospora crassa. in these experiments, procarbazine was a weak mutagen when present in growing cultures but nonmutagenic when conidial suspensions (nongrowing conidia) were treated. a total of 208 ad-3 mutants recovered after exposure of growing cultures of h-12 to 1 mg of procarbazine/ml, and 2 ad-3 mutants of spontaneous origin, were characterized genetic ...19911824718
relationship of the membrane atpase from halobacterium saccharovorum to vacuolar atpases.polyclonal antiserum against subunit a (67 kda) of the vacuolar atpase from neurospora crassa reacted with subunit i (87 kda) from a membrane atpase of the extremely halophilic archaebacterium halobacterium saccharovorum. the halobacterial atpase was inhibited by nitrate and n-ethylmaleimide; the extent of the latter inhibition was diminished in the presence of adenosine di- or triphosphates. 4-chloro-7-nitrobenzofurazan inhibited the halobacterial atpase also in a nucleotide-protectable manner; ...19911824911
cpc-1, the general regulatory gene for genes of amino acid biosynthesis in neurospora crassa, is differentially expressed during the asexual life cycle.cpci, the principal regulatory protein required for cross-pathway control of amino acid biosynthetic genes in neurospora crassa, contains a domain similar to the dna-binding domain of gcn4, the corresponding general regulator in saccharomyces cerevisiae. we examined binding by cpc1 synthesized in vitro and by cpc1 present in n. crassa whole-cell extracts. cpci from both sources was shown to bind to the dna sequence 5'-atgactcat-3', which is also the preferred recognition sequence of gcn4, cpc1 w ...19911824959
characterization of neurospora cpc1, a bzip dna-binding protein that does not require aligned heptad leucines for dimerization.cpc1 is the transcriptional activator of amino acid biosynthetic genes of neurospora crassa. cpc1 function in vivo was abolished upon deletion of segments of cpc-1 corresponding to the presumed transcription activation domain, the dna-binding and dimerization domains, or a 52-residue connector segment of cpc1. a truncated cpc1 polypeptide containing only the carboxy-terminal 57-residue segment of cpc1 was sufficient to form homodimers that bound dna. however, deletion of the segment of cpc-1 cor ...19911824960
catalysis of protein folding by cyclophilins from different species.cyclophilins are a class of ubiquitous proteins with yet unknown function. they were originally discovered as the major binding proteins for the immunosuppressant cyclosporin a. the only known catalytic function of these proteins in vitro is the cis/trans isomerization of xaa-pro bonds in oligopeptides. this became clear after the discovery that bovine cyclophilin is identical with porcine prolyl isomerase. this enzyme accelerates slow, proline-limited steps in the refolding of several proteins. ...19911825312
sequence similarities within the family of dihydrolipoamide acyltransferases and discovery of a previously unidentified fungal enzyme.a composite protein sequence database was searched for amino acid sequences similar to the c-terminal domain of the dihydrolipoamide acetyltransferase subunit (e2p) of the pyruvate dehydrogenase complex of escherichia coli. nine sequences with extensive similarity were found, of which eight were e2 subunits. the other was for a putative mitochondrial ribosomal protein, mrp3, from neurospora crassa. alignment of the mrp3 and e2 sequences showed that the similarity extends through the entire mrp3 ...19911825611
the neurospora crassa carotenoid biosynthetic gene (albino 3) reveals highly conserved regions among prenyltransferases.in the filamentous fungus neurospora crassa the biosynthesis of carotenoids is regulated by blue light. here we report the characterization of the albino-3 (al-3) gene of n. crassa, which encodes the carotenoid biosynthetic enzyme geranylgeranyl-pyrophosphate synthetase. this is the first geranylgeranyl-pyrophosphate synthetase gene isolated. nucleotide sequence comparison of al-3 genomic and cdna clones revealed that the al-3 gene is not interrupted by introns. transcription of the al-3 gene ha ...19911826006
identification of the active-site lysine residues of two biosynthetic 3-dehydroquinases.the lysine residues involved in schiff-base formation at the active sites of both the 3-dehydroquinase component of the pentafunctional arom enzyme of neurospora crassa and of the monofunctional 3-dehydroquinase of escherichia coli were labelled by treatment with 3-dehydroquinate in the presence of nab3h4. radioactive peptides were isolated by h.p.l.c. following digestion with cnbr (and in one case after further digestion with trypsin). the sequence established for the n. crassa peptide was alqh ...19911826831
all internal promoter elements of neurospora crassa 5 s rrna and trna genes, including the a boxes, are functionally gene-specific.the internal control elements of neurospora crassa 5 s genes include an a box and a c box as in xenopus and saccharomyces cerevisiae, plus a novel element, the ribo box at position +18 to +34. the ribo box is also found in the 40 s rrna promoter and a ribosomal protein gene but is absent from trna genes in n. crassa. the 5 s a box diverges from the trna a box consensus at two positions. we tested whether replacement of the 5 s a box with a trnaleu a box sequence would increase 5 s gene transcrip ...19911827115
sulfate transport in neurospora crassa: regulation, turnover, and cellular localization of the cys-14 protein.uptake of inorganic sulfate in neurospora crassa is governed by the sulfur regulatory circuit and is under the control of positively and negatively acting regulatory genes. two genetically and biochemically distinct systems are responsible for the uptake of sulfate from the environment. one of these, sulfate permease ii, encoded by the cys-14 gene, functions primarily in mycelia. a defined region of the cys-14 protein was highly expressed in escherichia coli and purified. anti-cys-14 antibody wa ...19911827594
duplication of leader sequence for protein targeting to mitochondria leads to increased import efficiency.we describe a novel method for enhancing protein import into mitochondria, by tandemly duplicating the n-terminal cleavable leader peptide using a gene manipulation strategy. the import into isolated yeast mitochondria of passenger proteins (yeast mitochondrial atp synthase subunits 8 and 9 and some mutagenised derivatives) that show little or no import when endowed with one such leader (that of neurospora crassa mitochondrial atp synthase subunit 9) is remarkably improved when the leader is tan ...19911828039
heterologous expression and regulation of the neurospora crassa nit-4 pathway-specific regulatory gene for nitrate assimilation in aspergillus nidulans.the nira gene of aspergillus nidulans and the nit-4 gene of neurospora crassa appear to be equivalent pathway-specific regulatory genes which mediate nitrate induction of nitrate reductase and nitrite reductase (nr and nir) activities. we have transformed the nit-4 wild-type (wt) gene into the a. nidulans loss-of-function (pleiotropic negative) nira 1 mutant strain. the nit-4 gene was found to complement the nira 1 mutation, thus permitting the nira 1 mutant strain to grow on nitrate or nitrite ...19911829047
one hour in 1 ata oxygen enhances rat alveolar macrophage chemiluminescence and fungal cytotoxicity.the purpose of this study was to determine if 100% o2 would enhance rat pulmonary alveolar macrophage (pam) oxidative killing of conidia of the fungus neurospora crassa. first, we found that incubation in 100% o2 had no effect on conidia viability in the absence of pam. we obtained resident pam from nonpretreated anesthetized male sprague-dawley rats by bronchoalveolar lavage. compared with similar air exposures we found that 1 h in vitro exposure of pam to 100% o2 (1.0 atmosphere absolute) incr ...19911829328
suppression of the cr-1 mutation in neurospora crassa.we have cloned a dna fragment, which hybridized with the adenylate cyclase gene (cyr1) of saccharomyces cerevisiae, from genomic dna libraries of neurospora crassa. the cr-1 mutation was able to be suppressed by introducing this dna fragment on a cosmid vector, judging from recovery of the adenylate cyclase activity and the abnormal morphology.19911829616
the beta-oxidation system in catalase-free microbodies of the filamentous fungus neurospora crassa. purification of a multifunctional protein possessing 2-enoyl-coa hydratase, l-3-hydroxyacyl-coa dehydrogenase, and 3-hydroxyacyl-coa epimerase activities.a trifunctional beta-oxidation protein, designated tfp, was purified to apparent homogeneity from oleate-induced mycelia of neurospora crassa. 2-enoyl-coa hydratase, l-3-hydroxyacyl-coa dehydrogenase, and 3-hydroxyacyl-coa epimerase activities copurified in constant ratios with this protein when crude extracts were subjected to cation-exchange, dye-ligand, and adsorption chromatography. trifunctionality was substantiated by coinciding enzyme activity ratios during the last two purification steps ...19911830048
the neurospora crassa cyt-20 gene encodes cytosolic and mitochondrial valyl-trna synthetases and may have a second function in addition to protein synthesis.the cyt-20-1 mutant of neurospora crassa is a temperature-sensitive, cytochrome b- and aa3-deficient strain that is severely deficient in both mitochondrial and cytosolic protein synthesis (r.a. collins, h. bertrand, r.j. lapolla, and a.m. lambowitz, mol. gen. genet. 177:73-84, 1979). we cloned the cyt-20+ gene by complementation of the cyt-20-1 mutation and found that it contains a 1,093-amino-acid open reading frame (orf) that encodes both the cytosolic and mitochondrial valyl-trna synthetases ...19911830127
nadh:ubiquinone oxidoreductase from bovine mitochondria. cdna sequence of a 19 kda cysteine-rich subunit.the sequence of a 19 kda subunit of nadh:ubiquinone oxidoreductase (complex i) from bovine heart mitochondria has been determined by a new strategy based on the polymerase chain reaction. the subunits of the enzyme were resolved in a polyacrylamide gel by two-dimensional isoelectric focusing and electrophoresis under denaturing conditions, transferred to a poly(vinylidene difluoride) membrane, and the n-terminal sequence was determined on the stained 19 kda protein up to residue 27. this informa ...19911830204
short dispersed repeats localized in spacer regions of chlamydomonas reinhardtii mitochondrial dna.in the mtdna of chlamydomonas reinhardtii, a unicellular green alga, we have identified a set of short repeated sequences up to 65 nucleotides long, each of which contains the palindromic consensus motif ctcgg(n4-14)ccgag. most of these repeated elements are localized in spacer regions that flank the transcribed coding regions of c. reinhardtii mtdna. these algal mitochondrial repeats have features reminiscent of short repeats in some fungal mtdnas, such as gc clusters in saccharomyces cerevisia ...19911831072
2-amino-n6-hydroxyadenine induces gene/point mutations and multiple-locus mutations, but not multilocus deletion mutations, in the ad-3 region of a two-component heterokaryon of neurospora crassa.the mutagenicity of 2-amino-n6-hydroxyadenine (aha) has been studied in neurospora crassa by treating a two-component heterokaryon (h-12) and recovering specific-locus mutations induced in the ad-3 region. this assay system permits the identification of ad-3a and/or ad-3b mutants resulting from gene/point mutations, multilocus deletion mutations, and multiple-locus mutations of various genotypes, involving one or both loci. genetic characterization of the ad-3 mutants recovered from experiments ...19911831243
cloning the mating types of the heterothallic fungus podospora anserina: developmental features of haploid transformants carrying both mating types.dnas that encode the mating-type functions (mat+ and mat-) of the filamentous fungus podospora anserina were cloned with the use of the mating-type a probe from neurospora crassa. cloning the full mat information was ascertained through gene replacement experiments. molecular and functional analyses of haploid transformants carrying both mating types lead to several striking conclusions. mat+ mat- strains are dual maters. however, the resident mat information is dominant to the mat information a ...19911831427
evolutionary conservation of a microbody targeting signal that targets proteins to peroxisomes, glyoxysomes, and glycosomes.peroxisomes, glyoxysomes, glycosomes, and hydrogenosomes have each been classified as microbodies, i.e., subcellular organelles with an electron-dense matrix that is bound by a single membrane. we investigated whether these organelles might share a common evolutionary origin by asking if targeting signals used for translocation of proteins into these microbodies are related. a peroxisomal targeting signal (pts) consisting of the cooh-terminal tripeptide serine-lysine-leucine-cooh has been identi ...19911831458
mutational analysis of the dna-binding domain of the cys3 regulatory protein of neurospora crassa.cys-3, the major sulfur regulatory gene of neurospora crassa, activates the expression of a set of unlinked structural genes which encode sulfur catabolic-related enzymes during conditions of sulfur limitation. the cys-3 gene encodes a regulatory protein of 236 amino acid residues with a leucine zipper and an upstream basic region (the b-zip region) which together may constitute a dna-binding domain. the b-zip region was expressed in escherichia coli to examine its dna-binding activity. the b-zi ...19911831537
primary structures of two subunits of nadh: ubiquinone reductase from neurospora crassa concerned with nadh-oxidation. relationship to a soluble nad-reducing hydrogenase of alcaligenes eutrophus.the primary structures of the nuclear-encoded 51 kda and 78 kda subunits of the respiratory chain nadh: ubiquinone reductase (complex i) from neurospora crassa mitochondria were determined by sequencing cdna and the n-terminus of the mature proteins. both subunits are related to the soluble nad-reducing hydrogenase of the bacterium alcaligenes eutrophus. sequence comparison between these subunits, the corresponding subunits of the bovine complex i and the bacterial nad-reducing hydrogenase furth ...19911832016
molecular organisation of the malate synthase genes of aspergillus nidulans and neurospora crassa.the sequencing and comparison of the genes encoding the glyoxylate bypass enzyme malate synthase of aspergillus nidulans (acue) and neurospora crassa (acu-9) are presented. the predicted amino acid sequences of the a. nidulans and n. crassa enzymes are 538 and 542 residues respectively and the proteins are 87% homologous. in fungi, the malate synthase proteins are located in glyoxysomes and the deduced acue and acu-9 proteins both contain a c-terminal s-k-l sequence, which has been implicated in ...19911832736
nadh:ubiquinone oxidoreductase from bovine heart mitochondria. cdna sequences of the import precursors of the nuclear-encoded 39 kda and 42 kda subunits.the 39 kda and 42 kda subunits of nadh:ubiquinone oxidoreductase from bovine heart mitochondria are nuclear-coded components of the hydrophobic protein fraction of the enzyme. their amino acid sequences have been deduced from the sequences of overlapping cdna clones. these clones were amplified from total bovine heart cdna by means of the polymerase chain reaction, with the use of complex mixtures of oligonucleotide primers based upon fragments of protein sequence determined at the n-terminals o ...19911832859
the initiation site for recombination cog is at the 3' end of the his-3 gene in neurospora crassa.recombination at his-3 in neurospora crassa is thought to be initiated through a site designated cog which lies in the his-3 to ad-3 interval of linkage group i. fragments of the his-3 gene were used to transform various his-3 mutant alleles to prototrophy in order to link the genetic map to the nucleotide sequence. it was established that cog is at the 3' end of his-3 and is therefore not the his-3 promoter. this suggests that cog may be dissimilar to a number of yeast recombinators which are a ...19911833619
antibodies against the 59 kda polypeptide of the n. crassa 8-10 nm filaments immunodetect a 59 kda polypeptide in specialized rat epithelial cells.p59nc is a polypeptide associated with bundles of cytoplasmic and nuclear filamentous structures of 8-10 nm of diameter in neurospora crassa cells. it is immunologically unrelated to both higher and lower eucaryotic tubulin and actin proteins and is detected weakly by the anti ifa monoclonal antibody. we analyze here the immunological relationship between p59nc and intermediate filament (if) mammalian proteins by using anti p59nc, anti keratin, anti vimentin and anti ifa antibodies. anti p59nc a ...19911833626
cloning and functional characterization of a eucaryotic dna photolyase gene from neurospora crassa.we cloned a genomic fragment of a photolyase gene from neurospora crassa by polymerase chain reaction using synthesized oligonucleotide primers designed from the most conserved amino acid sequences among photolyases of various organisms. using the cloned fragment as a hybridization probe we isolated a genomic fragment and cdna clones encoding the complete photolyase gene of this organism. the amino acid sequence of the photolyase deduced from the determined nucleotide sequence indicates a protei ...19911833725
[synthesis and antifungal activity of chlorobenzyl benzylidene thiazolidinediones and substituted of imidazolidinediones].the synthesis of six benzylidene thiazolidinediones and four benzylidene imidazolidinediones is described. in order to investigate their antifungal activity, they are evaluated against microorganism such as candida albicans, neurospora crassa, staphylococcus aureus and escherichia coli.19911834006
sequence of the nuclear atp synthase subunit 9 gene of podospora anserina: lack of similarity to the mitochondrial genome.the nuclear gene coding for the mitochondrial subunit 9 of the f0f1-atp synthase complex was isolated from a genomic library of podospora anserina. nucleotide sequencing revealed an open reading frame capable to code for 144 amino acids including an amino-terminal pre-sequence of 63 amino acid residues for mitochondrial import of the pre-proteolipid. the p. anserina proteolipid shows extensive sequence identity with the corresponding gene products of the related filamentous fungi neurospora cras ...19911834355
expression of meiotic drive elements spore killer-2 and spore killer-3 in asci of neurospora tetrasperma.it was shown previously that when a chromosomal spore killer factor is heterozygous in neurospora species with eight-spored asci, the four sensitive ascospores in each ascus die and the four survivors are all killers. sk-2k and sk-3k are nonrecombining haplotypes that segregate with the centromere of linkage group iii. no killing occurs when either one of these killers is homozygous, but each is sensitive to killing by the other in crosses of sk-2k x sk-3k. in the present study, sk-2k and sk-3k ...19911834522
utilization of the specific-locus assay in the ad-3 region of two-component heterokaryons of neurospora for risk assessment of environmental chemicals.the utilization of the specific-locus assay in the ad-3 region of two-component heterokaryons of neurospora crassa is compared with that of other eukaryotic assay systems for the evaluation of the mutagenic effects of environmental chemicals. in contrast to other in vitro specific-locus assays, the neurospora assay can detect mutations not only at the ad-3a and ad-3b loci but also recessive lethal mutations elsewhere in the genome. mutational damage in this system can be characterized readily by ...19911834935
qualitative differences in the spectra of genetic damage in 2-aminopurine-induced ad-3 mutants between nucleotide excision-repair-proficient and -deficient strains of neurospora crassa.the mutagenic effects of 2-aminopurine (2ap) have been compared 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). this forward-mutation, morphological and biochemical, specific-locus assay system permits the recovery of ad-3a and/or ad-3b mutants in 3 major classes: gene/point mutations, multilocus deletion m ...19911834936
characterization of two beta-tubulin genes from geotrichum candidum.the beta-tubulin genes g beta 1 and g beta 2 from the phytopathogenic hemiascomycete geotrichum candidum were found to be highly diverged in amino acid sequence from those of other filamentous fungi. g beta 1 and g beta 2 were also divergent from each other, with the coding regions sharing only 66% nucleotide sequence homology and 64% amino acid identity. however, the proteins shared 82% similarity and only 25 of the 161 non-identical amino acid substitutions were non-conservative. the organizat ...19911836049
chitin synthase 1 plays a major role in cell wall biogenesis in neurospora crassa.in filamentous fungi, chitin is a structural component of morphologically distinct structures assembled during various phases of growth and development. to investigate the role of chitin synthase in cell wall biogenesis in neurospora crassa, we cloned a chitin synthase structural gene and examined the consequences of its inactivation. using degenerate oligonucleotide mixtures designed on the basis of conserved sequences of the saccharomyces cerevisiae chs1 and chs2 polypeptides, a dna fragment e ...19911836444
coordinate expression of ribosomal protein genes in neurospora crassa and identification of conserved upstream sequences.the relative levels of rrnas and ribosomal proteins are coordinately regulated by growth rate and carbon nutrition in neurospora crassa. however, little is known about the mechanisms involved. to investigate the transcriptional regulation of ribosomal protein genes in n. crassa, we cloned and sequenced a ribosomal protein gene (crp-3). the inferred crp-3 protein sequence shares 89% and 83% homology at its n-terminus with the yeast rp51 and the human s17 ribosomal proteins respectively. the crp-3 ...19911836561
molecular cloning, characterization and analysis of the regulation of the aro2 gene, encoding chorismate synthase, of saccharomyces cerevisiae.we describe here the cloning, characterization and analysis of the regulation of the aro2 gene of saccharomyces cerevisiae, the first reported study of a eukaryotic gene encoding chorismate synthase (e.c. 4.6.1.4). the gene contains an orf of 1128 bp, encoding a protein with a calculated molecular mass of 40.8 kda. aro2 is regulated under the 'general control system' for amino acid biosynthesis by the transcriptional activator gcn4 which binds in vitro at three sites within the aro2 promoter. th ...19911837329
an electroporation-based system for high-efficiency transformation of germinated conidia of filamentous fungi.a rapid and efficient electroporation procedure has been developed for transformation of germinating conidia of filamentous fungi. pretreatment of conidial preparations with a cell wall weakening agent, such as beta-glucuronidase, was found to be essential for successful transformation. using the qa-2+ gene of neurospora crassa, encoding the catabolic dehydroquinase, as a selectable marker with a double-mutant host strain, auxotrophic for aromatic amino acids, integration of the plasmid was obse ...19911838030
cloning of a saccharomyces cerevisiae gene encoding a protein homologous to allantoicase of neurospora crassa. 19911839481
nit-4, a pathway-specific regulatory gene of neurospora crassa, encodes a protein with a putative binuclear zinc dna-binding domain.nit-4, a pathway-specific regulatory gene in the nitrogen circuit of neurospora crassa, is required for the expression of nit-3 and nit-6, the structural genes which encode nitrate and nitrite reductase, respectively. the complete nucleotide sequence of the nit-4 gene has been determined. the predicted nit4 protein contains 1,090 amino acids and appears to possess a single zn(ii)2cys6 binuclear-type zinc finger, which may mediate dna binding. site-directed mutagenesis studies demonstrated that c ...19911840634
nucleotide and derived amino acid sequences of the major porin of comamonas acidovorans and comparison of porin primary structures.the dna sequence of the gene which codes for the major outer membrane porin (omp32) of comamonas acidovorans has been determined. the structural gene encodes a precursor consisting of 351 amino acid residues with a signal peptide of 19 amino acid residues. comparisons with amino acid sequences of outer membrane proteins and porins from several other members of the class proteobacteria and of the chlamydia trachomatis porin and the neurospora crassa mitochondrial porin revealed a motif of eight r ...19911848840
the iron-sulfur clusters in the two related forms of mitochondrial nadh: ubiquinone oxidoreductase made by neurospora crassa.two related forms of the respiratory-chain complex, nadh: ubiquinone oxidoreductase (complex i) are synthesized in the mitochondria of neurospora crassa. normally growing cells make a large, piericidin-a-sensitive form, which consists of some 23 different nuclear- and 6-7 mitochondrially encoded subunits. cells grown in the presence of chloramphenicol make a small, piericidin-a-insensitive form which consists of only approximately 13 nuclear-encoded subunits. the subunits of the small form are e ...19911849820
cysteinyl-trna synthetase is a direct descendant of the first aminoacyl-trna synthetase.the gene encoding the cysteinyl-trna synthetase of e. coli was cloned from an e. coli genomic library made in lambda 2761, a lambda vector which can integrate and which carries a chloramphenicol resistance gene. a thermosensitive cyss mutant of e. coli was lysogenised and chloramphenicol-resistant colonies able to grow at 42 degrees c were selected to isolate phages containing the wild-type cyss gene. the sequence of the gene was determined. it codes for a 461 amino-acid protein and includes the ...19911864365
phylogenetic analysis of five medically important candida species as deduced on the basis of small ribosomal subunit rna sequences.the classification of species belonging to the genus candida berkhout is problematic. therefore, we have determined the small ribosomal subunit rna (srrna) sequences of the type strains of three human pathogenic candida species; candida krusei, c. lusitaniae and c. tropicalis. the srrna sequences were aligned with published eukaryotic srrna sequences and evolutionary trees were inferred using a matrix optimization method. an evolutionary tree comprising all available eukaryotic srrna sequences, ...19911865186
cloning, sequencing and expression of the schwanniomyces occidentalis nadp-dependent glutamate dehydrogenase gene.the cloned nadp-specific glutamate dehydrogenase (gdh) genes of aspergillus nidulans (gdha) and neurospora crassa (am) have been shown to hybridize under reduced stringency conditions to genomic sequences of the yeast schwanniomyces occidentalis. using 5' and 3' gene-specific probes, a unique 5.1 kb bcli restriction fragment that encompasses the entire schwanniomyces sequence has been identified. a recombinant clone bearing the unique bcli fragment has been isolated from a pool of enriched clone ...19911934128
oxidation of molybdopterin in sulfite oxidase by ferricyanide. effect on electron transfer activities.the attenuation of the sulfite:cytochrome c activity of sulfite oxidase upon treatment with ferricyanide was demonstrated to be the result of oxidation of the pterin ring of the molybdenum cofactor in the enzyme. oxidation of molybdopterin (mpt) was detected in several ways. ferricyanide treatment not only abolished the ability of sulfite oxidase to serve as a source of mpt to reconstitute the aponitrate reductase in extracts of the neurospora crassa mutant nit-1 but also eliminated the ability ...19912002036
identification of molybdopterin guanine dinucleotide in formate dehydrogenase from methanobacterium formicicum.the pterin cofactor in formate dehydrogenase isolated from methanobacterium formicium is identified as molybdopterin guanine dinucleotide. the pterin, stabilized as the alkylated, dicarboxamidomethyl derivative, is shown to have absorption and chromatographic properties identical to those of the previously characterized authentic compound. treatment with nucleotide pyrophosphatase produced the expected degradation products gmp and carboxyamidomethyl molybdopterin. the molybdopterin guanine dinuc ...19912037231
analysis of the altered mrna stability (ams) gene from escherichia coli. nucleotide sequence, transcriptional analysis, and homology of its product to mrp3, a mitochondrial ribosomal protein from neurospora crassa.the product of the altered mrna stability (ams) gene of escherichia coli is involved in decay of mrna. the complete nucleotide sequence of a 4-kilobase bamhi restriction fragment containing the ams coding sequence was determined. transcription of the ams gene was analyzed by high resolution s1 mapping. a promoter was found with a homology score of 58% 361 nucleotides upstream from the start codon of ams. the ams structural gene consists of an open reading frame of 2,445 nucleotides. the protein ...19911704367
voltage gating of the mitochondrial outer membrane channel vdac is regulated by a very conserved protein.soluble protein preparations obtained from the mitochondrial fractions of three very different organisms, neurospora crassa, rat, and potato, were discovered to greatly enhance the voltage sensitivity of the mitochondrial outer membrane channel, vdac. the active ingredient, referred to as the vdac modulator, increased the rate of voltage-dependent channel closure by approximately 10-fold. the modulator from one species increased the closing rate of vdac channels from all three species. the activ ...19911705100
localization of actin and characterization of its isoforms in the hyphae of neurospora crassa.the actin of neurospora crassa wild type strain st. lawrence has been purified, characterized and localized. a fungal 43 kda protein was isolated by affinity chromatography on dnase i-sepharose. this protein was identified as actin on immunoblots when an anti-actin monoclonal antibody raised against chicken gizzards was used as a probe. after two-dimensional gel electrophoresis three actin isoforms were detected. the distribution of actin in hyphae was examined by fitc-phalloidin staining of for ...19911706289
incipient mitochondrial evolution in yeasts. ii. the complete sequence of the gene coding for cytochrome b in saccharomyces douglasii reveals the presence of both new and conserved introns and discloses major differences in the fixation of mutations in evolution.we have determined the complete sequence of the mitochondrial gene coding for cytochrome b in saccharomyces douglasii. the gene is 6310 base-pairs long and is interrupted by four introns. the first one (1311 base-pairs) belongs to the group id of secondary structure, contains a fragment open reading frame with a characteristic giy ... yig motif, is absent from saccharomyces cerevisiae and is inserted in the same site in which introns 1 and 2 are inserted in neurospora crassa and podospora anseri ...19911708831
core i protein of bovine ubiquinol-cytochrome-c reductase; an additional member of the mitochondrial-protein-processing family. cloning of bovine core i and core ii cdnas and primary structure of the proteins.core i and core ii proteins are the largest nuclear-encoded subunits of the mitochondrial ubiquinol-cytochrome-c reductase (bc1 complex) lacking redox prosthetic groups. cdna clones of the two bovine core proteins have been isolated by the screening of lambda zap cdna libraries either with an oligonucleotide probe based on the sequence of an internal peptide or with a polymerase-chain-reaction-amplified fragment. the core i precursor protein consists of 362 amino acids with a 34-amino-acid prese ...19911712295
insulin-induced stimulation of protein phosphorylation in neurospora crassa cells.1) insulin stimulated the phosphorylation of at least 14 discrete proteins in neurospora crassa cells. specific proteins were phosphorylated at serine, threonine, and tyrosine residues, as determined by phosphoamino acid analysis of discrete spots on two-dimensional gels. 2) insulin stimulated the phosphorylation by [gamma-32p]atp of at least six discrete proteins in solubilized n. crassa membrane preparations at serine and tyrosine residues. 3) a phosphotyrosine-containing protein of 38 kda, pi ...19911717334
a nuclear gene with many introns encoding ammonium-inducible chloroplastic nadp-specific glutamate dehydrogenase(s) in chlorella sorokiniana.chlorella sorokiniana possesses ammonium-inducible, chloroplastic, nadp-specific glutamate dehydrogenase (nadp-gdh) homo- or heterohexamers composed of alpha- and/or beta-subunits which were previously shown to derive from precursor protein(s) of identical size. from the present studies, data are consistent with these two subunits being encoded by a single nuclear gene. the nadp-gdh gene is greater than 7 kb in length due to the presence of at least 21 introns, an unusually large number for a eu ...19911718478
specificity of leaf mitochondrial and chloroplast processing systems for nuclear-encoded precursor proteins.the specificity of the mitochondrial and chloroplast processing enzymes for the nuclear-encoded precursor proteins was investigated. mitochondrial precursor proteins of the nicotiana plumbaginifolia and the neurospora crassa beta subunits of f1-atpase and the neurospora rieske fes precursor protein were processed to the correct mature size by matrix extracts isolated from spinach leaves, yeast, rat liver and beef heart. the mitochondrial extracts failed to process chloroplast precursor proteins ...19911654154
calmodulin-dependent protein phosphatase from neurospora crassa. molecular cloning and expression of recombinant catalytic subunit.a cdna for the catalytic subunit of a calmodulin (cam)-dependent protein phosphatase was cloned from neurospora crassa. the open reading frame of 1557 base pairs encoded a protein of mr approximately 59,580 and was followed by a 3'-untranslated region of 363 base pairs including the poly(a) tail. based on primer extension analysis, the mrna transcript in vivo was 2403 base pairs. expression of this cam-protein phosphatase mrna was developmentally regulated, being highest during early mycelial gr ...19911655737
chiral linear hydroxamates as biomimetic analogues of ferrioxamine and coprogen and their use in probing siderophore-receptor specificity in bacteria and fungi.linear hydroxamate derivatives, possessing chiral alpha-amino acid moieties, were synthesized and their iron transport activities were studied in bacteria and fungi. no growth-promoting activity could be detected in the gram-positive hydroxamate-auxotroph aureobacterium flavescens jg9. however, gram-negative enterobacteria, such as escherichia coli, pantoea agglomerans and hafnia alvei were able to utilize iron from these analogues. uptake of 55fe-labeled analogues was inhibited by sodium azide, ...19911657086
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