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purification of a phosphatidylinositol/phosphatidylcholine transfer protein from neurospora crassa.this paper reports, for the first time, the purification of a phospholipid transfer protein (pltp) from a fungus, neurospora crassa. the protein was purified from the post-microsomal supernatant of n. crassa by successive chromatography on deae-cellulose, sephadex-g75 and pbe 94 (ph 4-7). the purified protein (m(r) 38,000) was found to transfer phosphatidylinositol preferentially over phosphatidylcholine, like the pltp from the yeast, saccharomyces cerevisiae. pc transfer was completely inhibite ...19921386256
arginine transport in mitochondria of neurospora crassa.transport of arginine into mitochondria of neurospora crassa has been studied. arginine transport was found to be saturable (km = 6.5 mm) and to have a ph optimum of ph 7.5. mitochondrial arginine transport appeared to be facilitated transport rather than active transport because: (i) the arginine concentration within the mitochondrial matrix after transport was similar to that of the reaction medium, and (ii) uncouplers and substrates of oxidative phosphorylation did not affect the transport ra ...19921386360
timing of synthesis and cellular localization of two conidiation-specific proteins of neurospora crassa.the process of conidiation in neurospora crassa consists of a series of distinct developmental stages culminating in the formation of multinucleate asexual spores called macroconidia. immunoblotting techniques were used to study the timing of synthesis and cellular localization of con10 and con13, the products of two genes that are expressed during conidiation but not during mycelial growth. both proteins first appear about 8 hr into conidiation; con10 disappears between 2 and 4 hr after germina ...19921386581
characterization and partial purification of an insulinase from neurospora crassa.an insulin-binding metal- and thiol-dependent proteinase has been purified 1491-fold from high speed cytosolic fractions of the fungus neurospora crassa. this enzyme resembles insulin-degrading enzymes (insulinases) present in mammalian cells and in drosophila melanogaster in the following ways: (i) it degrades radiolabeled insulin with a specificity similar to that of rat muscle insulinase, as demonstrated by hplc analysis of the degradation products; (ii) it is inhibited by bacitracin, edta, 1 ...19921386721
bioelectrorheological model of the cell. 3. viscoelastic shear deformation of the membrane.an analytical electromechanical model of a spherical cell exposed to an alternating electric field was used to calculate shear stress generated in the cellular membrane. shape deformation of neurospora crassa (slime) spheroplasts was measured. statistical analysis permitted empirical evaluation of creep of the cellular membrane within the range of infinitesimal stress. final results were discussed in terms of various rheological models.19921387010
electrophoretic profiles of mitochondrial plasmids in neurospora suggest they replicate by a rolling circle mechanism.migratory behaviour of mitochondrial plasmids from neurospora crassa mauriceville-1c and n. intermediate labelle has been studied by pulsed field gel electrophoresis (pfge). electrophoretic profiles demonstrate that long, linear molecules of a heterogeneous size are the prevailing form of plasmid dna in vivo. circular forms represent less than 8-9% of plasmid dna. single stranded dna regions are abundant and lead to electrophoretic inertia of a significant amount of plasmid dna. these profiles i ...19921387311
18o isotopic 13c nmr shift as proof that bifunctional peptidylglycine alpha-amidating enzyme is a monooxygenase.the biosynthesis of c-terminal alpha-amidated peptides from their corresponding c-terminal glycine-extended precursors is catalyzed by peptidylglycine alpha-amidating enzyme (alpha-ae) in a reaction that requires copper, ascorbate, and molecular oxygen. using bifunctional type a rat alpha-ae, we have shown that o2 is the source of the alpha-carbonyl oxygen of pyruvate produced during the amidation of dansyl-tyr-val-[alpha-13c]-d-ala, as demonstrated by the 18o isotopic shift in the 13c nmr spect ...19921387319
isolation and characterization of a new acetate-requiring mutant strain, ace-9, of neurospora crassa.a new acetate-requiring mutant strain of neurospora crassa, ace-9, has been isolated. the mutant gene was mapped between nuc-2 and arg-12 on the right arm of the second linkage group. the ace-9 mutant strain shows very weak activity of pyruvate dehydrogenase complex (pdhc). three strains that show no activity of pdhc had already been found, i.e., ace-2, ace-3, and ace-4. thus the ace-9 is the fourth gene that causes the deficiency in pdhc activity by a mutation. deficiency of pdhc activity in ac ...19921388033
a single amino-acid substitution in the beta-tubulin gene of neurospora confers both carbendazim resistance and diethofencarb sensitivity.two mbc-resistant mutants of neurospora crassa, f914 and f939, were sensitive to diethofencarb at a concentration of 0.1 micrograms/ml, while the wild-type strain and other mbc-resistant mutants showed resistance to diethofencarb at a concentration of 100 micrograms/ml. genetic analysis suggested that the mutations in these two strain were closely linked to the bml locus which codes for beta-tubulin. when the wild-type strain was transformed by the cloned beta-tubulin gene of the f914 strain, th ...19921388107
expression of neurospora crassa laccase under the control of the copper-inducible metallothionein-promoter.laccase from the ascomycete neurospora crassa is an inducible secretory enzyme. in vegetatively growing cultures its biosynthesis is repressed but can be induced by different protein synthesis inhibitors. transformation of the n. crassa wild-type strain singapore with a fusion gene consisting of the n. crassa copper-metallothionein promoter and the laccase gene are described in this report. correct integration of the 3.6 kilobase (kb) promoter-fragment fused with the laccase gene containing a 5' ...19921388108
transformants of neurospora crassa with the nit-4 nitrogen regulatory gene: copy number, growth rate and enzyme activity.nit-4 is a pathway-specific regulatory gene which controls nitrate assimilation in neurospora crassa, and appears to mediate nitrate induction of nitrate and nitrite reductase. the nit4 protein consists of 1090 amino-acid residues and possesses a single gal4-like putative dna-binding domain plus acidic, glutamine-rich, and polyglutamine regions. several mutants with amino-acid substitutions in the putative dna-binding domain and a nit-4 deletion mutant, which encodes a truncated nit4 protein lac ...19921388109
structure of the vacuolar atpase from neurospora crassa as determined by electron microscopy.we have examined the structure of the vacuolar atpase of neurospora crassa using negatively stained preparations of vacuolar membranes and of detergent-solubilized and gradient-purified atpase complexes. we also examined the peripheral sector (v1) of the enzyme after it had been removed and purified. using different stains, vacuolar membranes displayed ball-and-stalk structures similar to those of the intact mitochondrial atpase. however, the vacuolar atpase was clearly different from the mitoch ...19921388158
de novo synthesis and desaturation of fatty acids at the mitochondrial acyl-carrier protein, a subunit of nadh:ubiquinone oxidoreductase in neurospora crassa.we have cultivated the cel mutant of neurospora crassa defective in cytosolic fatty acid synthesis with [2-14c]malonate and found radioactivity covalently attached to the mitochondrial acyl-carrier protein (acp), a subunit of the respiratory chain nadh:ubiquinone oxidoreductase. we purified the acp by reverse-phase hplc: the bound acyl groups were trans-esterified to methylesters and analyzed by gas chromatography. the saturated c6 to c18 fatty acids and oleic acid were detected. de novo synthes ...19921397269
isolation of neurospora crassa a mating type mutants by repeat induced point (rip) mutation.in the filamentous fungus, neurospora crassa, mating type is regulated by a single locus with alternate alleles, termed a and a. the mating type alleles control entry into the sexual cycle, but during vegetative growth they function to elicit heterokaryon incompatibility, such that fusion of a and a hypha results in death of cells along the fusion point. previous studies have shown that the a allele consists of 5301 bp and has no similarity to the a allele; it is found as a single copy and only ...19921398049
characterization of the ilv-2 gene from neurospora crassa encoding alpha-keto-beta-hydroxylacyl reductoisomerase.we have isolated the cdna and corresponding genomic dna for the ilv-2 locus of neurospora crassa. this gene encodes alpha-keto-beta-hydroxylacyl reductoisomerase (ilv-2), required for the synthesis of isoleucine and valine. the gene contains four introns, maps to the right arm of chromosome v, and encodes a protein of 400 amino acids (aa). alignment of the aa sequence of ilv-2 with the two other known eukaryotic sequences encoding this enzyme reveals two conserved regions.19921398116
the cell division cycle gene cdc60 encodes cytosolic leucyl-trna synthetase in saccharomyces cerevisiae.the cdc60 mutation (for cell division cycle) of the yeast, saccharomyces cerevisiae, confers arrest at the start point of the cell cycle upon shift to the restrictive temperature [bedard et al., curr. genet. 4 (1981) 205-214]. we have cloned the cdc60 gene by complementation of the temperature-sensitive phenotype. sequence analysis revealed a single open reading frame of 3270 bp and the deduced amino acid sequence showed 50.5% sequence identity to the cytosolic leucyl-trna synthetase (leurs) fro ...19921398122
the vacuolar atpase of neurospora crassa.the filamentous fungus neurospora crassa has many small vacuoles which, like mammalian lysosomes, contain hydrolytic enzymes. they also store large amounts of phosphate and basic amino acids. to generate an acidic interior and to drive the transport of small molecules, the vacuolar membranes are densely studded with a proton-pumping atpase. the vacuolar atpase is a large enzyme, composed of 8-10 subunits. these subunits are arranged into two sectors, a complex of peripheral subunits called v1 an ...19921400281
phase resetting of the neurospora crassa circadian oscillator: effects of inositol depletion on sensitivity to light.the input pathway between the blue-light photoreceptor and the circadian oscillator of neurospora crassa has not yet been identified. to test the hypothesis that an inositol phospholipid signaling system might be involved in blue-light signal transduction, phase resetting by light was assayed in the inositol-requiring inl strain under conditions of inositol depletion. phase-resetting curves and dose-response curves indicated that cultures maintained on low inositol (25 microm) were several order ...20061421476
genetic organization and structural features of maranhar, a senescence-inducing linear mitochondrial plasmid of neurospora crassa.the nucleotide sequence of maranhar, a senescence-inducing linear mitochondrial plasmid of neurospora crassa, was determined. the termini of the 7-kb plasmid are 349-bp inverted repeats (tirs). each dna strand contains a long open reading frame (orf) which begins within the tir and extends toward the centre of the plasmid. orf-1 codes for a single-subunit rna polymerase that is not closely related to that encoded by another neurospora plasmid, kalilo. the orf-2 product may be a b-type dna polyme ...19921423726
length heterogeneity in its 2 and the methylation status of ccgg and gcgc sites in the rrna genes of the genus peronosclerospora.the polymerase chain reaction (pcr) was used with primers complementary to conserved flanking sequences to amplify the internal transcribed spacer 2 (its 2) of the rdna repeat units of five peronoscleropora isolates, one each of p. sorghi, p. maydis, p. sacchari and two of p. zeae. in contrast to the situation found in most-fungi that have been examined, length heterogeneity was evident in each sample. the rdna composition of the amplified bands was confirmed by southern hybridizations using an ...19921423729
development of a specific-locus assay in the ad-3 region of two-component heterokaryons of neurospora: a review.in recognition of the need for a more comprehensive data base for genetic risk assessment of human exposure to mutagenic agents in the environment, a model system was developed for specific-locus studies in neurospora crassa. this lower eukaryotic organism permits the utilization of microbial techniques for recovery of large numbers of specific-locus mutations at two closely linked loci as well as their subsequent genetic analysis. in particular, this assay makes possible exploratory experiments ...19921425606
characteristics of spontaneous and induced specific-locus mutation in the ad-3 region of neurospora crassa: utilization in genetic risk assessment.data from experiments on the induction of specific-locus mutations in model systems are utilized in genetic risk assessment to estimate potential adverse effects in the human population. in such assessments with radiation or chemical mutagens, the following information is required: (1) spontaneous and induced forward-mutation frequencies, (2) dose-response curves for the overall induction of specific-locus mutations, (3) genetic characterization of spontaneous and induced mutations, and (4) dose ...19921425607
immunocharacterization of actin and its immunofluorescent localization during the developmental gametophytic stages of allomyces arbuscula.highly specific polyclonal antibodies against actin from allomyces arbuscula were produced in rabbits, immunopurified by immunoblotting and specified with actin isolated from neurospora crassa and mouse skeletal muscle. used as immunofluorescence probes, they allowed localization of actin in the sequential gametophytic stages of the mould.19921426986
a combination inversion and translocation in neurospora crassa with inviable deficiency progeny that can be rescued in heterokaryons.chromosome rearrangement in(il;ir)t(il;iiir)slm-1, has a pericentric inversion in linkage group i associated with a reciprocal translocation between i and iii. the rearrangement was identified cytologically in pairing with normal sequence chromosomes at pachynema. rearrangement breakpoints were mapped genetically in il, ir and iiir by crosses with normal sequence strains and in crosses with an inversion that partially overlaps the slm-1 inversion. when rearrangement slm-1 is crossed to parents w ...19921427036
cytology of recessive sexual-phase mutants from wild strains of neurospora crassa.wild-collected strains of neurospora crassa harbor recessive mutations that are expressed in the sexual phase when homozygous. thirty-two representative mutants that produced barren perithecia were examined cytologically. six of these mutants failed to form asci. of the remaining 26, chromosome pairing was disturbed in 12 and meiosis was disturbed at pachytene or diplotene in 5. seven mutants showed normal meiosis i but then diverged from the normal sequence, and two showed perithecial beak abno ...19921427061
characterization of assembly intermediates of nadh:ubiquinone oxidoreductase (complex i) accumulated in neurospora mitochondria by gene disruption.nadh:ubiquinone oxidoreductase, the respiratory chain complex i of mitochondria, is an assembly of some 25 nuclear-encoded and 7 mitochondrially encoded subunits. the complex has an overall l-shaped structure formed by a peripheral arm and an elongated membrane arm. the peripheral arm containing one fmn and at least three iron-sulphur clusters constitutes the nadh dehydrogenase segment of the electron pathway. the membrane arm with at least one iron-sulphur cluster constitutes the ubiquinone red ...19921433284
sequence of the genes encoding subunits a and b of the vacuolar h(+)-atpase of schizosaccharomyces pombe.the genes encoding subunits a (vma1) and b (vma2) of the vacuolar h(+)-atpase from schizosaccharomyces pombe were cloned by hybridization to cdnas of the homologous genes in neurospora crassa. both genes are interrupted by introns, two in vma1 and four in vma2. positions of introns do not appear to be conserved when compared to those of n. crassa. the subunit a gene encodes a single product of 619 amino acids and is not interrupted by the coding sequence for a second product as found for sacchar ...19921441756
light and the recovery from heat shock induce the synthesis of 38 kda mitochondrial proteins in neurospora crassa.the effect of light on the protein synthesis pattern in the mitochondria of neurospora crassa was examined by in vivo labelling with [35s]-methionine and two-dimensional gel electrophoresis. a brief 5-min illumination induced the rapid and transient synthesis of a 38-kda protein. white collar-mutants were not stimulated to synthesize this protein by light. a protein of a similar molecular weight and isoelectrical point was synthesized during recovery from heat shock.19921444714
primary structure and mitochondrial import in vitro of the 20.9 kda subunit of complex i from neurospora crassa.the 20.9 kda subunit of nadh:ubiquinone oxidoreductase (complex i) from neurospora crassa is a nuclear-coded component of the hydrophobic arm of the enzyme. we have determined the primary structure of this subunit by sequencing a full-length cdna and a cleavage product of the isolated polypeptide. the deduced protein sequence is 189 amino acid residues long and contains a putative membrane-spanning domain. striking similarity over a 60 amino-acid-residue domain with the m (matrix) protein of par ...19921445273
identification of the ubiquinone-binding site of nadh:ubiquinone oxidoreductase (complex i) from neurospora crassa.in order to localize the ubiquinone-binding site of complex i (nadh:ubiquinone oxidoreductase), a novel photoreactive ubiquinone analogue (q0c7arn3) has been synthesized. it is shown that the direct chemical precursor of this analogue (q0c7arno2) and the analogue itself are accepted as substrates in an enzyme assay utilizing ubiquinone-depleted mitochondrial membranes of neurospora crassa. the activity of the enzyme applying these derivatives is inhibited by 50% at a concentration of 9 and 20 mi ...19751445878
characterization of the 9.5-kda ubiquinone-binding protein of nadh:ubiquinone oxidoreductase (complex i) from neurospora crassa.a small polypeptide subunit of the nadh:ubiquinone reductase (complex i) from neurospora crassa has been identified by photoaffinity labeling to participate in the binding of ubiquinone [heinrich, h., & werner, s. (1992) biochemistry (preceding paper in this issue)]. this polypeptide is further characterized by its primary structure and by an assessment of its localization within complex i. a lambda gt11 cdna expression library was screened using a specific antibody directed against this individ ...19921445879
prediction of transmembrane topology of f0 proteins from escherichia coli f1f0 atp synthase using variational and hydrophobic moment analyses.the a subunit, a membrane protein from the e. coli f1f0 atp synthase has been examined by fourier analysis of hydrophobicity and of amino-acid residue variation. the amino-acid sequences of homologous subunits from vibrio alginolyticus, saccharomyces cerevisiae, neurospora crassa, aspergillus nidulans, schizosaccharomyces pombe and candida parapsilosis were used in the variability analysis. by fourier analysis of sequence variation, two transmembrane helices are predicted to have one face in con ...19921445940
cloning and sequencing of the cdna encoding tyrosinase of the japanese pond frog, rana nigromaculata.we cloned and sequenced the cdna encoding tyrosinase (tyn) of the japanese pond frog, rana nigromaculata. the 3511-bp cdna contained a 54-bp 5'-noncoding region, a 1596-bp open reading frame encoding tyn of 532 amino acids (aa), and a 1861-bp 3'-noncoding region. the aa sequence of frog tyn predicted from the cdna sequence was homologous to that of mouse and human tyns. the aa sequence including the copper-binding domain, which is likely the active center of tyn, was highly conserved among these ...19921446833
photoinduction of albino-3 gene expression in neurospora crassa conidia.the synthesis of carotenoids is induced by blue light in neurospora crassa mycelia, while in conidia (the vegetative spores) the accumulation of carotenoids also occurs in the dark. the expression of the albino-3 (al-3) gene (coding for the carotenogenic enzyme geranyl-geranyl pyrophosphate synthetase) in isolated conidia was analysed. the level of al-3 mrna was shown to be increased in light-induced wild type (wt) conidia. this light response was elicited by blue light and was under the control ...19921453275
developmental and light regulation of eas, the structural gene for the rodlet protein of neurospora.the surface of many fungal spores is covered by a hydrophobic sheath termed the rodlet layer. we have determined that the rodlet protein of neurospora crassa is encoded by a cloned gene designated bli-7, and that bli-7 is identical to the known gene eas (easily wettable). using eas dna as a probe we show that eas mrna is abundant in illuminated mycelia and conidiophores but is not detectable or is barely detectable in dark-grown mycelia, mature macroconidia, microconidia, and ascospores. mutatio ...19921459459
the neurospora circadian clock-controlled gene, ccg-2, is allelic to eas and encodes a fungal hydrophobin required for formation of the conidial rodlet layer.the neurospora crassa clock-controlled gene (ccg-2) is transcriptionally activated by the circadian clock in a time-of-day-specific manner. transcript and sequence analyses of ccg-2 reveal that the predicted ccg-2 polypeptide bears significant similarity to a class of low-molecular-weight, cysteine-rich, hydrophobic proteins (hydrophobins), first identified in schizophyllum, and including the product of the developmentally regulated aspergillus gene, rodletless, required for spore surface rodlet ...19921459460
probing the structure of the neurospora crassa plasma membrane h(+)-atpase.the structure of the neurospora crassa plasma membrane h(+)-atpase has been investigated using a variety of chemical and physiochemical techniques. the transmembrane topography of the h(+)-atpase has been elucidated by a direct, protein chemical approach. reconstituted proteoliposomes containing purified h(+)-atpase molecules oriented predominantly with their cytoplasmic surface facing outward were treated with trypsin, and the numerous peptides released were purified by hplc and subjected to am ...19921461258
the regulatory gene nit-2 of neurospora crassa complements a nnu mutant of gibberella zeae (fusarium graminearum).the nnu mutant of gibberella zeae (=fusarium graminearum) is unable to catabolize many of the nitrogen sources utilized by its wild-type parent, and may have suffered a mutation in the major nitrogen regulatory locus. transformation of this mutant with the major nitrogen regulatory gene from neurospora crassa, nit-2, restored the wild-type phenotype, thus confirming that the nnu mutation is in the major nitrogen regulatory locus of g. zeae. our results are consistent with the premise of conserva ...19921465117
[synthesis and structure of substituted bromo and nitrobenzyl benzylidene imidazolidinediones and thiazolidinediones].synthesis and physico-chemical properties of five bromobenzyl-benzylidene-imidazolidinediones and five nitrobenzyl- or benzyl-benzylidene-thiazolidinediones are described. the microbiological activity of bromobenzyl-benzylidene-imidazolidinediones against microorganisms such as candida albicans, neurospora crassa and mycobacterium smegmatis are evaluated.19921471825
isolation, partial amino acid sequence, and cellular distribution of heat-shock protein hsp98 from neurospora crassa.hsp98 is one of the most prominent proteins synthesized during the heat-shock response of neurospora crassa. we purified hsp98 and determined the amino acid sequence of two overlapping peptides obtained by cyanogen bromide cleavage. this 28 amino acid sequence from hsp98 has 75% homology with a region of the clpb protein of escherichia coli and 86% homology to a 96-kda protein of trypanosoma brucei. it also has 71% homology to hsp104 of saccharomyces cerevisiae. hsp98 was enriched in the microso ...19921472534
neurospora crassa blue-light-inducible gene bli-7 encodes a short hydrophobic protein.blue light induces a number of physiological reactions in neurospora crassa. we have cloned and sequenced the gene bli-7, which is inducible by blue light, and both glucose and nitrogen starvation. this gene is strongly expressed at the rna-level and contributes up to 0.2% of n. crassa total rna when fully induced. the deduced amino acid sequence reveals a short (108 amino acids), hydrophobic protein which has homology to the protein sc-3, encoded by a gene of schizophyllum commune which was iso ...19921472707
quelling: transient inactivation of gene expression in neurospora crassa by transformation with homologous sequences.up to 36% of neurospora crassa transformants showing an albino phenotype were recovered by transforming a wild-type strain with different portions of the carotenogenic albino-3 (al-3) and albino-1 (al-1) genes. the presence of the exogenous sequences (which were randomly integrated in ectopic locations) provoked a severe impairment in the expression of the endogenous al-1 or al-3 genes. this phenomenon, which we have termed 'quelling', was found to be spontaneously and progressively reversible, ...20061484489
further characterization of the histidine gene cluster of streptomyces coelicolor a3(2): nucleotide sequence and transcriptional analysis of hisd.we have further characterized the genomic region of streptomyces coelicolor a3(2) that contains genes involved in the biosynthesis of histidine. a 2,357-base pair fragment contained in plasmid psch3328 that complemented hisd mutations has been sequenced. computer analysis revealed an open reading frame that encodes a protein with significant homology to the escherichia coli, salmonella typhimurium and mycobacterium smegmatis hisd product, saccharomyces cerevisiae his4c, and neurospora crassa his ...19921488552
vacuolar atpase of neurospora crassa: electron microscopy, gene characterization and gene inactivation/mutation.we are using three approaches to investigate the vacuolar atpase, v-atpase, from neurospora crassa. (1) examination in the electron microscope shows the enzyme has a 'ball and stalk' structure like the f-type atpases. however, the vacuolar atpase is significantly larger, has a prominent cleft in the head sector, and has extra components associated with the stalk and membrane sectors. (2) genes encoding three of the major subunits of the vacuolar atpase and the homologous subunits of the mitochon ...19921491233
a dominant selectable marker that is meiotically stable in neurospora crassa: the amds gene of aspergillus nidulans.when neurospora crassa is transformed using a neurospora gene as the selectable marker, the vegetatively stable transformants obtained cannot be used successfully in a cross because the selectable marker will be inactivated by the process of rip (repeat-induced point mutation). introduction of the acetamidase-encoding gene amds of aspergillus nidulans into n. crassa by transformation yielded transformants that would grow in minimal medium containing acetamide as a sole nitrogen source. in mitoti ...19921494342
molecular cloning and characterization of the saccharomyces cerevisiae cyt2 gene encoding cytochrome-c1-heme lyase.cytochrome c1, a subunit of the mitochondrial ubiquinol--cytochrome-c reductase, is synthesized on cytosolic ribosomes as a precursor protein of 37 kda. maturation to the mature 31-kda form involves two proteolytic processing steps of the amino-terminal presequence. after removal of the amino-terminal part by the matrix-localized processing peptidase, the carboxy-terminal part of the presequence is cleaved off by an unknown intermembrane space protease. this step depends on covalent linkage of h ...19921499554
efficient transformation of claviceps purpurea using pyrimidine auxotrophic mutants: cloning of the omp decarboxylase gene.a homologous transformation system was developed for the phytopathogenic fungus claviceps purpurea. orotidine-5'-monophosphate decarboxylase (ompd)-deficient mutants were obtained by uv mutagenesis and selection for resistance against 5-fluoroorotate. these mutants could be complemented well by the corresponding genes of aspergillus niger (pyra) and neurospora crassa (pyr4), yielding significantly higher transformation rates (and lower copy numbers per transformant) than the phleomycin resistanc ...19921508154
sequences of 20 subunits of nadh:ubiquinone oxidoreductase from bovine heart mitochondria. application of a novel strategy for sequencing proteins using the polymerase chain reaction.nadh:ubiquinone oxidoreductase, the first enzyme in the respiratory electron transport chain of mitochondria, is a membrane-bound multi-subunit assembly, and the bovine heart enzyme is now known to contain about 40 different polypeptides. seven of them are encoded in the mitochondrial dna; the remainder are the products of nuclear genes and are imported into the organelle. the primary structures of 12 of the nuclear coded subunits have been described and those of a further 20 are described here. ...19921518044
ornithine decarboxylase gene of neurospora crassa: isolation, sequence, and polyamine-mediated regulation of its mrna.ornithine decarboxylase (odc), which initiates the biosynthesis of the polyamines putrescine, spermidine, and spermine, is encoded by the spe-1 gene of the fungus neurospora crassa. this gene and its cdna have been cloned and sequenced. the gene has a single 70-nucleotide intron in the coding sequence. the cdna, comprising the entire coding region, recognizes a single 2.4-kb mrna in northern (rna) blots. the mrna transcript, defined by s1 mapping, has an extremely long, 535-base leader without s ...19921530878
the mitochondrial tyrosyl-trna synthetase of podospora anserina is a bifunctional enzyme active in protein synthesis and rna splicing.the neurospora crassa mitochondrial tyrosyl-trna synthetase (mt tyrrs), which is encoded by the nuclear gene cyt-18, functions not only in aminoacylation but also in the splicing of group i introns. here, we isolated the cognate podospora anserina mt tyrrs gene, designated yts1, by using the n. crassa cyt-18 gene as a hybridization probe. dna sequencing of the p. anserina gene revealed an open reading frame (orf) of 641 amino acids which has significant similarity to other tyrrss. the yts1 orf i ...19921531084
nit2, the nitrogen regulatory protein of neurospora crassa, binds upstream of nia, the tomato nitrate reductase gene, in vitro.the nit-2 gene of neurospora crassa encodes a trans-acting regulatory protein that activates the expression of a number of structural genes which code for nitrogen catabolic enzymes, including nitrate reductase. the nit2 protein contains a cys2/cys2-type zinc-finger dna-binding domain that recognizes promoter regions of the neurospora nitrogen-related genes. the nit2 zinc-finger domain/beta-gal fusion protein was shown to recognize and bind in a specific manner to two upstream fragments of the n ...19921531184
characterization of the glyoxysomal isocitrate lyase genes of aspergillus nidulans (acud) and neurospora crassa (acu-3).the nucleotide sequences of the genes encoding the acetate-inducible glyoxylate cycle enzyme isocitrate lyase from the ascomycete fungi aspergillus nidulans (acud) and neurospora crassa (acu-3) are presented. the respective a. nidulans and n. crassa genes are interrupted at identical positions by two introns and encode proteins of 538 and 543 amino acids, which have 75% identity. the predicted protein sequences do not demonstrate the c-terminal tripeptide s-k-l that has been implicated in peroxi ...19921531185
glvr1, a receptor for gibbon ape leukemia virus, is homologous to a phosphate permease of neurospora crassa and is expressed at high levels in the brain and thymus.the human gene glvr1 has been shown to render mouse cells sensitive to infection by gibbon ape leukemia virus. this indication that the glvr1 protein acts as a virus receptor does not reveal the protein's normal physiological role. we now report that glvr1 is homologous to pho-4+, a phosphate permease of neurospora crassa, at a level sufficiently high to predict that glvr1 is also a transport protein, although the substrate transported remains unknown. to characterize the gene further, we have c ...19921531369
molecular characterization of mutations of nit-4, the pathway-specific regulatory gene which controls nitrate assimilation in neurospora crassa.the nit-4 genes of three conventional neurospora crassa mutations and of the closely related species, neurospora intermedia, have been isolated by amplifying the genomic dna with the polymerase chain reaction. nucleotide sequencing has revealed that the three nit-4 mutants, alleles 15, 1214, and 2994, are the result of a missense mutation, a nonsense mutation and a frameshift mutation, respectively. the nucleotide sequence of the nit4 protein coding region of a nit-4 mutant (allele 2994) and of ...19921531376
sequence and characterization of the met-7 gene of neurospora crassa.the proteins encoded by the met-7+ and met-3+ genes of neurospora crassa are required to form a functional cystathionine-gamma-synthase (cgs). the met-7+ gene has been cloned by complementation of a met-7 mutant. the nucleotide sequence of the complementing dna reveals the presence of a 542-amino acid open reading frame (orf). disruption of this orf abolishes complementation of the met-7 mutation.19921531800
characterization of the formate (for) locus, which encodes the cytosolic serine hydroxymethyltransferase of neurospora crassa.serine hydroxymethyltransferase (shmt) occupies a central position in one-carbon (c1) metabolism, catalyzing the reaction of serine and tetrahydrofolate to yield glycine and 5,10-methylenetetrahydrofolate. methylenetetrahydrofolate serves as a donor of c1 units for the synthesis of numerous compounds, including purines, thymidylate, lipids, and methionine. we provide evidence that the formate (for) locus of neurospora crassa encodes cytosolic shmt. the for+ gene was localized to a 2.8-kb bglii f ...19921532227
production of the cys3 regulator, a bzip dna-binding protein, is sufficient to induce sulfur gene expression in neurospora crassa.the cys-3+ gene of neurospora crassa encodes a bzip (basic region-leucine zipper) regulatory protein that is essential for sulfur structural gene expression (e.g., ars-1+). nuclear transcription assays confirmed that cys-3+ was under sulfur-regulated transcriptional control and that cys-3+ transcription was constitutive in sulfur controller (scon)-negative regulator mutants. given these results, i have tested whether expression of cys-3+ under high-sulfur (repressing) conditions was sufficient t ...19921532230
effects of light on protein secretion in neurospora crassa.the relative concentrations of secreted proteins in liquid cultures of neurospora crassa differ in constant darkness compared to constant light (2500 lx). light reduces the concentrations of some polypeptides markedly and increases the concentrations of protein species of 67, 40, 18 and 13 kda. the "blind" wc-2 mutant of neurospora does not show light dependent differences in amounts of secreted proteins. one of the light-sensitive extracellular proteins is shown to be a protease of 17.5 kda.19921532303
the dna-binding domain of the cys-3 regulatory protein of neurospora crassa is bipartite.cys-3, the major sulfur regulatory gene of neurospora crassa, encodes a regulatory protein that is capable of sequence-specific interaction with dna. the interaction is mediated by a region within the cys3 protein (the bzip region) which contains a potential dimer-forming surface, the leucine zipper, and an adjacent basic dna contact region, nh2-terminal to the leucine zipper. to investigate the bipartite nature of the bzip region, a series of cys-3 mutants obtained by oligonucleotide-directed m ...19921532511
use of gene replacement transformation to elucidate gene function in the qa gene cluster of neurospora crassa.gene replacement by transformation, employing selective genetic recombination techniques, has been used to delete or disrupt the qa-x, qa-y and qa-1s genes of the qa gene cluster of neurospora crassa. the growth characteristics of the strain carrying the deletion of the qa-y gene support earlier evidence that this gene encodes a quinic acid permease. the strain containing the deletion of the qa-1s gene (delta qa-1s) was examined with respect to quinic acid induction and carbon catabolite repress ...19921533844
analysis of junction sequences resulting from integration at nonhomologous loci in neurospora crassa.we have analyzed the junctions involved in two examples of ectopic integration of plasmids containing the am+ (glutamate dehydrogenase) gene into a strain of neurospora crassa bearing a complete deletion of the am locus. in one transformed strain a single copy of plasmid dna had been integrated into linkage group (lg) iii dna without the loss of chromosomal dna. in contrast, 450 bp had been lost from plasmid sequences at the site of integration. the transforming dna used was circular, so we post ...19921533845
saccharomyces cerevisiae and neurospora crassa contain heavy metal sequestering phytochelatin.in fungi, cellular resistance to heavy metal cytotoxicity is mediated either by binding of metal ions to proteins of the metallothionein type or by chelation to phytochelatin-peptides of the general formula (gamma-glu-cys)n-gly. hitherto, only one fungus, candida glabrata has been shown to contain both metal inactivating systems. here we show by unambiguous fab-ms analysis that both a metallothionein-free mutant of saccharomyces cerevisiae as well as a wildtype strain synthesize phytochelatin (p ...19921534214
expression of con genes along the three sporulation pathways of neurospora crassa.the filamentous fungus neurospora crassa produces three types of spores by using different developmental pathways: macroconidiation, microconidiation, and sexual spore (ascospore) formation. several genes of unknown function have been cloned by virtue of their expression during macroconidiation but not during mycelial growth (con genes). it had been postulated that expression of the con genes was specific to macroconidiation. to test this assumption, protein extracts from macroconidia, microconi ...19921534304
new polyenic antibiotics active against gram-positive and gram-negative bacteria. viii. construction of synthetic medium for production of mono-chloro-congeners of enacyloxins.new antibiotics enacyloxins (enxs) are a family of non-lactonic polyene antibiotics produced by frateuria sp. w-315. for the production of antibiotics, we had to employ two-step fermentations, the first is the production of spent medium of neurospora crassa and the second is the production of antibiotics by frateuria. to simplify the production of antibiotics, systematic analyses have been done on the spent medium, and factors which affect the production of antibiotics characterized. from the ab ...19921534321
cot-1, a gene required for hyphal elongation in neurospora crassa, encodes a protein kinase.neurospora crassa is a filamentous fungus that grows on semisolid media by forming spreading colonies. mutations at several loci prevent this spreading growth. cot-1 is a temperature sensitive mutant of n.crassa that exhibits restricted colonial growth. at temperatures above 32 degrees c colonies are compact while at lower temperatures growth is indistinguishable from that of the wild type. restricted colonial growth is due to a defect in hyphal tip elongation and a concomitant increase in hypha ...19921534751
the cleavable presequence is not essential for import and assembly of the phosphate carrier of mammalian mitochondria but enhances the specificity and efficiency of import.the phosphate carrier (pic) of mammalian mitochondria is synthesized with a cleavable presequence, in contrast to other members of the mitochondrial family of inner membrane carrier proteins. the precursor of pic is efficiently imported, proteolytically processed, and correctly assembled in isolated mitochondria. here we report that a presequence-deficient pic was imported with an efficiency of about 50% as compared with the authentic precursor of pic. this mature-sized pic was correctly assembl ...19921534805
the mating types of podospora anserina: functional analysis and sequence of the fertilization domains.the two idiomorphic alleles called mat+ and mat-, which control the mating types in podospora anserina, have been cloned. mat+ and mat- encompass 3.8 kb and 4.7 kb respectively, of unrelated dna sequences flanked by common sequences. subcloning allowed the identification and localization in each locus of the gene that controls fertilization, probably by determining the mating type. the mat+ gene, called fpr1, encodes a protein with a potential dna-binding hmg domain. the presence of this motif s ...19921534866
alternative modes of mrna processing in a 3' splice site mutant of neurospora crassa.the am8 mutant of neurospora crassa is shown to have a double base-pair change, gtg for the normal tag, at the 3' end of the second intron of the am (nadp-specific glutamate dehydrogenase, gdh) gene. the greater part of the mutant am transcript accumulates as two fragments hybridising to probes for sequences respectively upstream and downstream of the 5' end of the intron. two processed transcripts approximating to normal full length mrna were identified. in one the second intron was intact; in ...19921535288
circadian rhythms in neurospora crassa: the role of mitochondria.energy metabolism and mitochondria have been discussed with respect to their role in the circadian rhythm mechanism for some time. numerous examples of inhibitors that affect the mitochondria of plants and animals and microorganisms are known, which cause large phase shifts in the rhythms of these organisms. analogous studies on the role of mitochondria in the neurospora circadian rhythm mechanism have also been reported and summarized. this communication differs from previous studies on other o ...19921535289
molecular analysis of the neurospora clock: cloning and characterization of the frequency and period-4 genes.genetic analysis of neurospora crassa has identified many mutants that affect the biological clock. in this article we review the cloning of two of these genes, frq and prd-4. both genes were isolated using a chromosome walk technique. subcloning experiments and subsequent northern analysis of frq implicate the importance of two transcripts that emanate from this locus. in preliminary data, no protein-coding region is evident in the smaller transcript; the larger transcript contains a 962-amino ...19921535290
molecular analysis of the laccase gene from the chestnut blight fungus and selective suppression of its expression in an isogenic hypovirulent strain.the gene encoding laccase in the chestnut blight fungus, cryphonectria parasitica, has been cloned and characterized. the predicted c. parasitica laccase amino acid sequence (591 aa) was 57% identical to the neurospora crassa laccase sequence and contained four potential copper-binding regions that are conserved in a number of copper-binding proteins. treatment of a virulent c. parasitica strain with 3 microm cycloheximide resulted in a marked increase in laccase mrna accumulation, whereas ident ...20081535523
control of mating type heterokaryon incompatibility by the tol gene in neurospora crassa and n. tetrasperma.the mating-type of neurospora crassa (a and a) have a dual function: a and a individuals are required for sexual reproduction, but only strains of the same mating type will form a stable vegetative heterokaryon. neurospora tetrasperma, in contrast, is a naturally occurring a+a heterokaryon. it was shown previously that the mating-type genes of both species are functionally the same and are not responsible for this difference in heterokaryon incompatibility. this suggests that a separate genetic ...19921535606
cloning and sequence analysis of a rapamycin-binding protein-encoding gene (rbp1) from candida albicans.rapamycin (rm) and fk506 are macrolide antifungal agents that exhibit potent immunosuppressive properties in higher eukaryotes which are mediated through interaction with specific receptor proteins (fkbps or rbps, for fk506- and rm-binding proteins, respectively). these proteins possess peptidyl-prolyl cis-trans isomerase (ppiase) activity in vitro which is inhibited by the binding of rm and fk506. we previously isolated a gene encoding an rbp from saccharomyces cerevisiae, and demonstrated that ...19921563628
small subunit ribosomal rna of blastomyces dermatitidis: sequence and phylogenetic analysis.we determined the small subunit (18s) ribosomal rna sequence of the dimorphic fungus blastomyces dermatitidis. the sequence was compared to that of fourteen other eukaryotic organisms, ten of which were higher fungi, and an evolutionary tree was constructed based on these sequences. b. dermatitidis aligned most closely with the ascomycetes neurospora crassa and podospora anserina, in agreement with previous phylogenetic analysis based on morphological criteria. phase-specific cdna clones derived ...19921564447
[synthesis and antimicrobial activity of chlorobenzyl benzylidene imidazolidinedione derivatives and substituted thiazolidinediones].the synthesis of five chlorobenzyl benzylidene imidazolidinediones and four fluorobenzyl benzylidene thiazolidinediones is described. in order to investigate their antimicrobial activity they are evaluated against microorganism such as candida albicans, neurospora crassa, staphylococcus aureus, mycobacterium smegmatis and escherichia coli.19921615023
identification of a family of bacteriophage t4 genes encoding proteins similar to those present in group i introns of fungi and phage.the bacteriophage t4 sega gene lies in a genetically unmapped region between the gene beta gt (beta-glucosyltransferase) and uvsx (recombination protein) and encodes a protein of 221 amino acids. we have found that the first 100 amino acids of the sega protein are highly similar to the n termini of four other predicted t4 proteins, also of unknown function. together these five proteins, sega-e (similar to endonucleases of group i introns), contain regions of similarity to the endonuclease i-tev ...19921631169
direct transfer of molybdopterin cofactor to aponitrate reductase from a carrier protein in chlamydomonas reinhardtii.a chlamydomonas reinhardtii molybdenum cofactor (moco)-carrier protein (cp), capable of reconstituting nitrate reductase activity with apoprotein from the neurospora crassa mutant nit-1, was subjected to experiments of diffusion through a dialysis membrane and gel filtration. cp bonded firmly moco and did not release it efficiently unless aponitrate reductase was present in the incubation mixture. stability of moco bound to cp against air and heat was very similar to that of free-moco released f ...19921644169
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
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
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
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
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
alteration of the cytochrome c oxidase subunit 2 gene in the [exn-5] mutant of neurospora crassa.the maternally inherited [exn-5] mutant of neurospora crassa is characterized by its slow-growth rate and deficiency of cytochrome aa3 relative to wild-type strains. we have determined the dna sequence of the coxi and coxii genes of the mutant, which encode subunits 1 and 2 of cytochrome c oxidase, respectively. no changes in the dna sequence of the coxi gene relative to the corresponding wild-type gene were found. in the region of the coxii gene we found two alterations, one a c to t transition ...19911657411
over-expression, purification and determination of the proteolytic processing site of the yeast mitochondrial cbs1 protein.yeast transformants harboring the cbs1 gene under the control of the strong adc1 promoter on a high copy number plasmid express the mitochondrial cbs1 protein at artificially high levels. over-expressed protein is imported into mitochondria and correctly processed to yield the mature mitochondrial 23.5 kda form, but differs in its solubility properties from cbs1 in wild-type mitochondria. it forms insoluble protein aggregates, which are refractory to solubilization with 1% taurodeoxycholate. we ...19911657414
a new cyclitol derivative influences inositol metabolism in neurospora crassa.cyclitol derivatives have been synthesized and screened for growth inhibitory effect upon prokaryotic and eukaryotic organisms. one derivative, (2s,3r,5r)-3-azido-2-benzoyloxy-5-hydroxycyclohexanone, was studied in detail: it has no effect upon bacteria, but it is inhibitory to neurospora crassa. in neurospora crassa it increased the amount of myo-inositol-1-phosphate synthase and inhibited the activity of myo-inositol-monophosphatase. the enhanced synthesis of myo-inositol-1-phosphate synthase ...19911657695
ubiquitination of endogenous calmodulin in rabbit tissue extracts.previously we were able to show that purified calmodulins from vertebrates, plants (spinach) and the mold neurospora crassa can be covalently conjugated to ubiquitin in a ca(2+)-dependent manner. it was therefore pertinent to answer the question if a tissue extract contains all the components necessary for the endogenous synthesis of ubiquityl calmodulin (ucam). therefore [125i]ubiquitin, atp/mg2+ and ca2+ were added to tissue extracts enriched by a single ion exchange step. in such extracts of ...19911661685
molecular comparison of the negative-acting nitrogen control gene, nmr, in neurospora crassa and other neurospora and fungal species.in neurospora crassa, the expression of unlinked structural genes which encode nitrogen catabolic enzymes is subject to genetic and metabolic regulation. the negative-acting nmr regulatory gene appears to play a role in nitrogen catabolite repression. using the n. crassa nmr gene as a probe, homologous sequences were identified in a variety of other filamentous fungi. the polymerase chain reaction was used to isolate the nmr-like gene from the exotic mauriceville strain of n. crassa and from the ...19911663340
in vitro inhibition of stable 1,3-beta-d-glucan synthase activity from neurospora crassa.glucan synthase activity of neurospora crassa was isolated by treatment of protoplast lysates with 0.1% 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate and 0.5% octylglucoside in 25 mm 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer, ph 7.4, containing 5 mm edta, 1 mm phenylmethylsulfonylfluoride, 200 mm inorganic phosphate, 10 microm gtp, 1 mm dtt, 10 mm sodium fluoride, and 600 mm glycerol. resulting activity was partially purified by sucrose gradient density sedimentation ...19911669437
isolation and characterization of the adenylate cyclase structural gene of neurospora crassa.a single gene (nac) encoding an adenylate cyclase was cloned from the genomic dna library of neurospora crassa, using the dna fragment encoding the catalytic domain of adenylate cyclase of saccharomyces cerevisiae as a probe. the open reading frame of this gene (6900 base pairs) was interrupted three time by introns. the protein encoded consists of 2300 amino acids and has adenylate cyclase activity. n. crassa adenylate cyclase has a high degree of homology with the catalytic domains of yeast an ...19911680356
genomic analysis of a virulent and a less virulent strain of the entomopathogenic fungus beauveria bassiana, using restriction fragment length polymorphisms.the genomic dna of two strains of the entomopathogenic fungus beauveria bassiana, strain gk2016, a "wild type" (virulent), and strain gk2051, a less virulent mutant to grasshoppers, was digested with 12 restriction endonucleases. gel electrophoresis conditions were established to show restriction fragment length patterns visually in the digested dna stained with ethidium bromide. the less virulent mutant was generated by ultraviolet illumination of conidiospores at a 95% lethal dose. both strain ...19911680543
isolation and expression of the acetate-inducible isocitrate lyase gene (acu-3) from neurospora crassa: evidence for a second constitutive isozyme.heterologous hybridisation of the aspergillus nidulans structural gene for isocitrate lyase (acud) to a lambda genomic library of neurospora crassa identified a recombinant phage containing the hybridising sequence on an internal 9 kb ecori fragment. a restriction fragment length polymorphism (rflp) enabled the fragment to be assigned to linkage group v (lg v), the location of the acetate-inducible isocitrate lyase, acu-3 of neurospora. functional ectopic complementation by co-transformation of ...19911681413
sequence and expression of gln3, a positive nitrogen regulatory gene of saccharomyces cerevisiae encoding a protein with a putative zinc finger dna-binding domain.the gln3 gene of saccharomyces cerevisiae is required for the activation of transcription of a number of genes in response to the replacement of glutamine by glutamate as source of nitrogen. we cloned the gln3 gene and constructed null alleles by gene disruption. gln3 is not essential for growth, but increased copies of gln3 lead to a drastic decrease in growth rate. the complete nucleotide sequence of the gln3 gene was determined, revealing one open reading frame encoding a polypeptide of 730 a ...19911682800
comparative studies of the quinic acid (qa) cluster in several neurospora species with special emphasis on the qa-x-qa-2 intergenic region.the organization of the quinic acid (qa) genes in neurospora crassa has been compared to that in several other neurospora species. this gene cluster was found to be highly conserved in all species examined. however, there are numberous restriction fragment length polymorphisms that distinguish the heterothallic and homothallic species. catabolic dehydroquinase assays indicated that qa-2 gene expression in the homothallic species is subject to induction by quinic acid, as is the case in n. crassa ...19911685010
isolation and sequence of an fk506-binding protein from n. crassa which catalyses protein folding.slow protein-folding reactions are accelerated by a prolyl cis/trans isomerase isolated from porcine kidney which is identical to cyclophilin, a protein that is probably the cellular receptor for the immunosuppressant cyclosporin a. catalysis probably involves the isomerization of prolyl peptide bonds in the folding protein chains. cyclosporin a inhibits folding catalysis by cyclophilin. here we report the isolation, cloning, sequencing and expression of another protein with prolyl isomerase act ...19901696687
optimized vectors and selection for transformation of neurospora crassa and aspergillus nidulans to bleomycin and phleomycin resistance.to provide a dominant selectable marker for transformation of neurospora crassa strains lacking specific auxotrophic mutations, we have engineered the bleomycin (bm) resistance-encoding gene (ble) from the bacterial transposon tn5 for expression in n. crassa. the coding region of the ble gene was fused to the promoter and terminator regions of the n. crassa am gene. in some vectors, multiple cloning sites were placed flanking the ble gene to provide a versatile ble cassette. when introduced into ...19901699844
the apocytochrome b gene of chlamydomonas smithii contains a mobile intron related to both saccharomyces and neurospora introns.the mitochondrial dna of the two interfertile algal species chlamydomonas smithii and chlamydomonas reinhardtii are co-linear with the exception of ca. 1 kb insertion (the alpha insert) present in c. smithii dna only. in vegetative diploids resulting from interspecific crosses, mitochondrial genomes are transmitted biparentally except for the alpha insert which is transmitted to all c. reinhardtii molecules in a manner reminiscent of the intron-mediated conversion event that occurs at the omega ...19901701210
nucleotide sequence of the exons of the large subunit rrna of neurospora crassa mitochondria. 19901701879
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
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