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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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
comparison of the spectra of genetic damage in n4-hydroxycytidine-induced ad-3 mutations between nucleotide excision repair-proficient and -deficient heterokaryons of neurospora crassa.a comparison has been made of the mutagenic effects of n4-hydroxycytidine (hc) in the adenine-3 (ad-3) region of two-component heterokaryons of neurospora crassa: nucleotide excision repair-proficient (uvs-2+/uvs-2+) heterokaryon 12 (h-12) and nucleotide excision repair-deficient (uvs-2/uvs-2) heterokaryon 59 (h-59). hc was found to produce mutations predominantly, if not exclusively, by at to gc base-pair transitions in escherichia coli strain k12 by janion and glickman (1980, mutation res., 72 ...19937678887
nucleotide sequence of the large mitochondrial rrna gene of penicillium chrysogenum.the nucleotide sequence of a large rrna (1-rrna) gene and its flanking regions in the cloned fragments of mitochondrial (mt) dna from penicillium chrysogenum nrrl1951 (sekiguchi j., ohsaki, t., yamamoto, h., koichi, k. and shida, t. (1990) j. gen. microbiol. 136, 535-543) was determined and compared with those in aspergillus nidulans and neurospora crassa mitochondrial dnas. the p. chrysogenum mt 1-rrna gene has a 1678 bp intron which intervenes between a 2835 bp 5' exon and a 581 bp 3' exon, an ...19937680578
mei-3, a recombination and repair gene of neurospora crassa, encodes a reca-like protein.neurospora crassa mei-3 is a mutant which exhibits meiotic and mitotic defects and mutagen sensitivity. its defect is believed to be in recombination and repair. we have cloned the mei-3 gene from a n. crassa cosmid library of genomic dna. restriction fragment length polymorphism analysis determined the location of the cloned fragment was on chromosome one in approximately the same position that was previously reported for mei-3 by classical genetic methods. deletion analysis showed the approxim ...19937692262
design of an object-oriented database for reverse genetics.we present the design of an object-oriented database system for reverse genetics applications. such a database will encapsulate not only the data in the genetic and physical maps, but also the methods used to create the maps as well as methods to link them to other databases, such as genbank, pir, and medline. the purpose of this database is to provide the fungal genetics community with an electronic tool for identifying the biochemical function of any dna fragment in the database--electronic re ...19937584341
use of aspergillus overproducing mutants, cured for integrated plasmid, to overproduce heterologous proteins.aspergillus niger var. awamori was previously transformed with a vector designed to express a fused glucoamylase-prochymosin gene and bearing the neurospora crassa pyr4 gene as a selectable marker. mutant strains that overproduced the glucoamylase-prochymosin fusion protein were derived from one of the transformants. despite the fact that the expression vector was integrated into the genome of these strains it was possible to obtain strains from which the vector sequences had been removed. this ...19937764120
development of a new transformant selection system for penicillium chrysogenum: isolation and characterization of the p. chrysogenum acetyl-coenzyme a synthetase gene (faca) and its use as a homologous selection marker.a new transformation system for the filamentous fungus penicillium chrysogenum is described, based on the use of the homologous acetyl-coenzyme a synthetase (faca) gene as a selection marker. acetate-non-utilizing (fac-) strains of p. chrysogenum were obtained by positive selection for spontaneous resistance to fluoroacetate. among these fac mutants putative faca strains were selected for a loss of acetyl-coenzyme a (coa) synthetase activity. the faca gene, coding for the enzyme acetyl-coa synth ...19937765289
structure and sequence of the calmodulin gene from neurospora crassa.cdna and genomic clones of neurospora calmodulin were obtained by pcr. characterization revealed an open reading frame encoding a predicted protein of 149 amino acids, showing 85% identity to the human calmodulin protein sequence. comparison of the cdna and genomic sequence reveals the position of five introns, organized differently than is found in calmodulin genes from higher eukaryotes.19937916628
cloning and sequence analysis of an h(+)-atpase-encoding gene from the human dimorphic pathogen histoplasma capsulatum.a gene related to the pma1 gene from saccharomyces cerevisiae was isolated from the pathogenic human dimorphic fungus, histoplasma capsulatum, using fungal-specific oligodeoxyribonucleotide (oligo) probes. this gene has been given the name hc-pma1. the structural organization of hc-pma1 consists of three exons (375, 2329 and 44 bp) and two introns (115 and 116 bp). the nucleotide sequence predicts an h(+)-atpase-related protein of 916 amino acids (aa). comparison of the deduced aa sequence to th ...19937916725
molecular cloning, characterization, and nucleotide sequence of nit-6, the structural gene for nitrite reductase in neurospora crassa.the neurospora crassa assimilatory nitrite reductase structural gene, nit-6, has been isolated. a cdna library was constructed from poly(a)+ rna isolated from neurospora mycelia in which nitrate assimilation had been induced. this cdna was ligated into lambda zap ii (stratagene) and amplified. this library was then screened with a polyclonal antibody specific for nitrite reductase. a total of six positive clones were identified. three of the six clones were found to be identical via restriction ...19938096840
the neurospora uvs-2 gene encodes a protein which has homology to yeast rad18, with unique zinc finger motifs.a clone containing the dna repair gene uvs-2 of neurospora crassa was identified from a neurospora genomic dna library using the sib-selection method. transformants were screened for resistance to methyl methane sulfonate (mms). a dna fragment that complements the uvs-2 mutation was subcloned by monitoring its ability to transform the uvs-2 mutant to mms resistance. deletion analysis of the cloned dna indicated that the size of the uvs-2 gene is approximately 1.6 kb. the identity of the uvs-2 ge ...19938097557
a series of yeast/escherichia coli lambda expression vectors designed for directional cloning of cdnas and cre/lox-mediated plasmid excision.a series of saccharomyces cerevisiae/escherichia coli lambda/plasmid expression vectors have been constructed which allow easy excision of the plasmid sequences from lambda. features of six are described, and two designated lambda pg15 and lambda ad5, are characterized in detail. transcription of cloned sequences is controlled by the alternative promoters, adh2, pgk, gal10 and sv40 early, and by the cyc1 transcriptional terminator. unique ecori and xhoi restriction sites in the intervening polyl ...19938154182
functional cooperation of mitochondrial protein import receptors in yeast.we have identified a 20 kda yeast mitochondrial outer membrane protein (termed mas20) which appears to function as a protein import receptor. we cloned, sequenced and physically mapped the mas20 gene and found that the protein is homologous to the mom19 import receptor from neurospora crassa. mas20 and mom19 contain the sequence motif f-x-k-a-l-x-v/l, which is repeated several times with minor variations in the mas70/mom72 receptors. to determine how mas20 functions together with the previously ...19938223428
higher plant mitochondria encode an homologue of the nuclear-encoded 30-kda subunit of bovine mitochondrial complex i.we describe the structure and expression of a wheat mitochondrial gene, which codes for a subunit of mitochondrial nadh dehydrogenase. the deduced protein sequence has 70% similarity to the 30-kda subunit of bovine mitochondrial complex i and 65% similarity to the 31-kda subunit of neurospora crassa complex i, components of the iron-sulfur-protein fraction, both nuclear-encoded proteins. we named this wheat mitochondrial gene as nad9. the wheat nad9 gene is transcribed in a single mrna of 0.9 kb ...19938223639
identification of the heat shock protein of neurospora crassa corresponding to the stress-inducible peroxidase.heat shock and other stress treatments, resulting in thermotolerance in neurospora crassa cells, stimulate the induction of a peroxidase at a high level. the putative gene encoding this heat shock-induced peroxidase (hspp) has been cloned, using a cdna clone of the manganese peroxidase of phanerochaete chrysosporium, as a probe. northern blot analysis of total rna from heat-shocked cell showed the stress-dependent accumulation of a approximately 10 kb transcript. the identity of the hsp, corresp ...19938240345
de novo methylation of repeated sequences in coprinus cinereus.we have examined the stability of duplicated dna sequences in the sexual phase of the life cycle of the basidiomycete fungus, coprinus cinereus. we observed premeiotic de novo methylation in haploid nuclei containing either a triplication, a tandem duplication, or an ectopic duplication. methylation changes were not observed in unique sequences. repeated sequences underwent methylation changes during the dikaryotic stage. in one cross, 27% of the segregants exhibited methylation-directed gene in ...19938244000
the sugar beet mitochondrial genome contains an orf sharing sequence homology with the gene for the 30 kda subunit of bovine mitochondrial complex i.from a sugar beet mitochondrial dna library, we have isolated an open reading frame (orf192) showing extensive homology to the gene for the 30 kda subunit of the bovine mitochondrial complex i (nadh: ubiquinone reductase). the orf192 was found to be actively transcribed to give an rna of approximately 1.0 kb. we have designated this gene nad9. transcripts from the nad9 locus are edited by five c to u transitions, increasing similarity with the amino acid sequence of the corresponding bovine and ...19938246903
characterization of the 3-dehydroquinase domain of the pentafunctional arom protein, and the quinate dehydrogenase from aspergillus nidulans, and the overproduction of the type ii 3-dehydroquinase from neurospora crassa.the arom protein of aspergillus nidulans is a multidomain pentafunctional polypeptide that is active as a dimer and catalyses steps 2-6 in the prechorismate section of the shikimate pathway. the three c-terminal domains (including the type i 3-dehydroquinase) of the arom protein are homologous with the qutr-encoded qutr protein that represses transcription of the eight genes comprising the quinic acid utilization (qut) gene cluster, and the two n-terminal domains are homologous with the quta-enc ...19938257437
higher-plant cofactor-independent phosphoglyceromutase: purification, molecular characterization and expression.cofactor-independent phosphoglyceromutase (pgm) was purified to homogeneity from developing castor seed endosperm. immunological characterization using monospecific antisera raised against this protein indicates that the enzyme is located in the cytosol and that there is no immunologically related polypeptide in the leucoplast from this tissue. isolation and sequence determination of full-length cdna clones for castor and tobacco pgm demonstrate that the protein is highly conserved in these plan ...19938260624
effect of site-directed mutagenesis of conserved aspartate and arginine residues upon farnesyl diphosphate synthase activity.all polyprenyl synthases catalyze the condensation of the allylic substrate, isopentenyl diphosphate, with a specific homoallylic diphosphate substrate. polyprenyl synthases from homo sapiens, ratus rattus, escherichia coli, saccharomyces cerevisiae, neurospora crassa, and erwinia herbicola contain two conserved "aspartate-rich domains" (ashby, m.n., and edwards, p.a. (1992) j. biol. chem. 267, 4128-4136). in order to determine the importance of these domains in catalysis, the conserved aspartat ...19938262934
pas10 is a tetratricopeptide-repeat protein that is essential for the import of most matrix proteins into peroxisomes of saccharomyces cerevisiae.pas mutants of saccharomyces cerevisiae are disturbed in peroxisome assembly (pas) and proliferation. here we report the characterization of the pas10 gene and its product (pas10) that is essential for the import of a large subset of proteins into the peroxisomal matrix. pas10, a protein of 69 kda, is a member of the tetratricopeptide repeat, or snap helix, protein family, characterized by several direct repeats of a degenerate 34-amino acid motif (sikorski, r. s., boguski, m. s., goebl, m. & hi ...19938265627
an escherichia coli gene showing a potential ancestral relationship to the genes for the mitochondrial import site proteins isp42 and mom38.an orf (orft) of 1911 base pairs, upstream of the hip operon in escherichia coli at map position 33.82 has been identified. the protein encoded by this sequence is predicted to have a molecular mass of 68,249 da and the carboxyterminal 276 residues shows 26.8% and 25.4% identity with the import site proteins isp42 and mom38 from the mitochondrial outer membrane of saccharomyces cerevisiae and neurospora crassa, respectively. these mitochondrial membrane proteins have been shown to be essential c ...19938274505
the positive-acting sulfur regulatory protein cys3 of neurospora crassa: nuclear localization, autogenous control, and regions required for transcriptional activation.the positive-acting global sulfur regulatory protein, cys3, of neurospora crassa turns on the expression of a family of unlinked structural genes that encode enzymes of sulfur catabolism. cys3 contains a leucine zipper and an adjacent basic region (b-zip), which together constitute a bipartite sequence-specific dna-binding domain. specific anti-cys3 antibodies detected a protein of the expected size in nuclear extracts of wild-type neurospora under conditions in which the sulfur circuit is activ ...19938316209
choline: its role in the growth of filamentous fungi and the regulation of mycelial morphology.choline is an essential metabolite for the growth of filamentous fungi. it occurs most notably as a component of the major membrane phospholipid, phosphatidyl choline (lecithin), and fulfills a major role in sulphate metabolism in the form of choline-o-sulphate in many species. choline is usually synthesised endogenously, but exogenous choline can also be taken up, either to compensate for metabolic deficiencies in choline-requiring mutants such as those of aspergillus nidulans and neurospora cr ...19938318261
identification of a g protein alpha subunit from neurospora crassa that is a member of the gi family.heterotrimeric g proteins, consisting of alpha, beta, and gamma subunits, are implicated in major signal transduction pathways controlling a diversity of functions in eukaryotic organisms. in the filamentous fungus neurospora crassa, g proteins are implicated in the regulation of several environmental responses. as a first step in studying the role of g proteins in these processes, we have cloned the genes for two alpha subunits, gna-1 and gna-2, from neurospora. the genes are located on differe ...19938325859
calcium and microorganisms.this review followed from experiments suggesting that some fungi do not require calcium. it was found that many studies of a calcium requirement in microorganisms had assumed specificity for chelation agents such as egta and a23187, which the reagents did not possess. early studies still cited today often preceded the recognition that microorganisms required manganese and zinc. as a result of both of these misunderstandings, there was rarely any attempt to replace calcium by other important trac ...19938338620
heterologous expression of mating-type genes in filamentous fungi.podospora anserina and neurospora crassa, two filamentous heterothallic ascomycetes, have a single mating-type locus with two alternate forms called mat+ and mat- and a and a, respectively. mating type controls entry into the sexual cycle, events subsequent to fertilization, and, in n. crassa, prevents the formation of mixed mating-type heterokaryons. the mating types of these two organisms display similarity in their dna structure and in the encoded polypeptides involved in fertilization. here ...19938341677
functional identification of al-3 from neurospora crassa as the gene for geranylgeranyl pyrophosphate synthase by complementation with crt genes, in vitro characterization of the gene product and mutant analysis.in this work the neurospora crassa al-3 gene function was determined. geranylgeranyl pyrophosphate (ggpp) synthase activity was measured in al-2 fgsc 313 and al-3 rp100 fgsc 2082 mutant strains by in vitro synthesis methods. this experiment showed that al-3 rp100 mutant expresses a reduced ggpp synthase activity. the mutated al-3 gene was cloned and sequenced; a single missense mutation was found changing serine into asparagine. genetic complementation was performed by escherichia coli transform ...19938350190
cloning of the gene for glutamate decarboxylase and its expression during conidiation in neurospora crassa.neurospora crassa glutamate decarboxylase (gad) is produced during conidiation and stored in dormant conidia. polyclonal antibody was generated to gad that had been purified to homogeneity. the anti-gad antibody was specific for n. crassa gad and inhibited gad activity. the level of gad protein decreased during conidial germination, indicating that gad was degraded during this phase of development. the anti-gad antibody was used to isolate a cdna clone of gad from a lambda zap cdna expression li ...19938352739
is an activator protein-2-like transcription factor involved in regulating gene expression during nitrogen limitation in fungi?the upstream sequences of all published lignin peroxidase and manganese peroxidase genomic clones from phanerochaete chrysosporium were analyzed. this analysis revealed the presence of putative activator protein-2 (ap-2) recognition sequences in 11 of 15 lignin peroxidase genes. the lignin peroxidase clone glg6 and the manganese peroxidase gene (mnp-1) have two copies of putative ap-2 sequence in the upstream region. interestingly, the lignin peroxidase gene vlg4 of another white rot fungus, tra ...19938357266
functional domains of the transcriptional activator nuc-1 in neurospora crassa.the nuc-1 regulatory protein directly controls the transcription of these genes and how the activity enzymes in neurospora crassa. to understand how nuc-1 regulates the transcription of these genes and how the activity of nuc-1 is modulated by other regulatory proteins, two putative functional domains of nuc-1 were analysed: the dna-binding domain and the regulatory domain. the dna-binding activity of nuc-1 has not been directly demonstrated; however, results of deletion analysis, sequence analy ...19938359693
purification and characterization of the assimilatory nitrate reductase of azotobacter vinelandii.1. a soluble reduced methyl viologen-dependent assimilatory nitrate reductase from azotobacter vinelandii strain uw136 grown aerobically on nitrate was purified to homogeneity by the criteria of nitrate reductase activity staining, and coincidence of a coomassie blue-staining protein band on polyacrylamide gels run under non-denaturing conditions. the specific activity was 3 mumol of no2- formed/min per mg of protein. 2. gel filtration on superose-12 and sds/page showed that the enzyme had an m( ...19938380991
the sta2 and mel1 genes of saccharomyces cerevisiae are idiomorphic.the sta2 (glucoamylase) gene of saccharomyces cerevisiae has been mapped close to the end of the left arm of chromosome ii. meiotic analysis of a cross between a haploid strain containing sta2, and another strain carrying the melibiase gene mel1 (which is known to be at the end of the left arm of chromosome ii) produced parental ditype tetrads only. since there is no significant dna sequence similarity between the sta2 and mel1 genes, or their respective flanking regions, we conclude that these ...19938381338
interchangeable associations of calcineurin regulatory subunit isoforms with mammalian and fungal catalytic subunits.two mammalian genes for the ca(2+)-binding regulatory subunit of the calmodulin-dependent protein phosphatase (calcineurin) were identified recently, suggesting unique associations with tissue-specific catalytic subunits. the murine brain (beta 1) and testis (beta 2) isoforms of the regulatory subunit were expressed with poly-histidine carboxyl termini and purified by ni(2+)-chelate chromatography, each exhibiting high affinity ca2+ binding on nitrocellulose overlays. using chromatographic metho ...19938384215
mitochondrial cardiolipin in diverse eukaryotes. comparison of biosynthetic reactions and molecular acyl species.cardiolipin, a unique dimeric phospholipid of bacteria and mitochondria, can be synthesized by two alternative pathways discovered in rat and escherichia coli, respectively. in mitochondrial preparations from fungi (saccharomyces cerevisiae, neurospora crassa), higher plants (phaseolus aureus), molluscs (mytilus edulis) and mammals (rat liver, bovine adrenal gland), cardiolipin was synthesized from cdp-diacylglycerol and phosphatidylglycerol, suggesting a common eukaryotic mechanism of cardiolip ...19938385010
multiple isomers of phosphatidyl inositol monophosphate and inositol bis- and trisphosphates from filamentous fungi.the range of inositol phosphates and inositol phospholipids present in three filamentous fungi, neurospora crassa, fusarium graminearum and phanerochaete chrysosporium has been investigated by hplc analysis. the profiles obtained demonstrate that two isomers of phosphatidyl inositol monophosphate are present, and that an apparent complexity in the number of isomers of inositol bis- and trisphosphates is found in filamentous fungi that has not been observed in animal or plant cells.19938394259
glycogen-bound protein in lower eukaryote and prokaryote.the proteoglycogen fraction of neurospora crassa was purified and subjected to radioiodination with [125i]iodide. amylolysis of the polysaccharide moiety led to the isolation of a labelled 31 kda-protein. the nh2-terminal amino acid sequence of 10 residues of the 31 kda-protein was determined. a 31 kda-protein was also bound to glycogen in escherichia coli. proteoglycogen has not been heretofore found in any primitive unicellular organism.19938401303
complete coding sequence, intron/exon organization, and chromosomal location of the gene for the core i protein of human ubiquinol-cytochrome c reductase.core i protein is a nuclear-encoded component of the ubiquinol-cytochrome c reductase complex of the mitochondrial respiratory chain. we have located the gene for the human core i protein in the p21 region of chromosome 3, just upstream of the col7a1 gene which encodes type vii collagen. the core i gene, which has been sequenced in its entirety, is comprised of 10,417 base pairs, from the major transcription start site to the polyadenylation signal, and contains 13 exons. the predicted polypepti ...19938407948
a tobacco cdna clone encoding a gata-1 zinc finger protein homologous to regulators of nitrogen metabolism in fungi.in higher plants, the expression of the nitrate assimilation pathway is highly regulated. although the molecular mechanisms involved in this regulation are currently being elucidated, very little is known about the trans-acting factors that allow expression of the nitrate and nitrite reductase genes which code for the first enzymes in the pathway. in the fungus neurospora crassa, nit-2, the major nitrogen regulatory gene, activates the expression of unlinked structural genes that specify nitroge ...19938413186
molecular cloning of the smaller subunit(p52) of rat liver mitochondrial processing protease.a cdna encoding the smaller subunit (p52) of mitochondrial processing protease was isolated from a rat liver cdna library, using cdna fragment for yeast mas1 as the probe. the deduced amino acid sequence is highly homologous to those of pep from neurospora crassa and mas1 from saccharomyces cerevisiae. after in vitro transcription and translation, the precursor peptide was imported into isolated rat liver mitochondria and processed to its mature form.19938422255
insertional mutagenesis in neurospora crassa: cloning and molecular analysis of the preg+ gene controlling the activity of the transcriptional activator nuc-1.the transcriptional activator nuc-1 controls the transcription of the genes for phosphorus acquisition enzymes, and its activity is regulated by the negative regulatory factors, preg and pgov in this report, we describe the cloning and molecular analysis of the preg+ gene. in neurospora crassa, as in higher eukaryotes, transformation frequently results in nonhomologous integration of transforming dna. insertion of transforming dna into host genes mutates the gene and provides a molecular tag for ...19938436269
the mitochondrial genome of the entomopathogenic fungus beauveria bassiana: analysis of the ribosomal rna region.the 28.5-kbp mitochondrial (mt) genome from the entomopathogenic fungus beauveria bassiana was studied using restriction enzyme analysis, gene probe hybridization, and dna sequence comparisons. a detailed restriction enzyme map allowed cloning of the entire genome into a number of segments. hybridization of heterologous gene probes to the mtdna resulted in the identification of the large ribosomal rna (lrrna) and small ribosomal rna (srrna) genes. gene probes derived from several yeasts and fung ...19938439871
primary structure of the nuclear-encoded 10.5 kda subunit of complex i from neurospora crassa.we have isolated a cdna clone for the nuclear encoded 10.5 kda subunit of complex i from n. crassa. dna sequencing revealed an open reading frame corresponding to a polypeptide with 94 amino acids and a calculated molecular mass of 10531 da. the protein is synthesized without a cleavable mitochondrial targeting sequence. the n. crassa polypeptide is the fungal equivalent of subunit b8 of bovine complex i.19938448210
peptide splicing in the vacuolar atpase subunit a from candida tropicalis.subunit a of the vacuolar proton pump appears to be responsible for the atp hydrolysis which is coupled to the pumping of protons into a variety of intracellular acid compartments, including the fungal vacuole. we report here the cloning and sequence determination of the gene encoding subunit a from candida tropicalis. southern blot hybridization analysis indicates that there is a single gene which encodes this protein. the gene contains a single intron at the extreme 5'-end of the coding region ...19938463270
isolation, sequencing, and characterization of crp-5, a gene encoding a neurospora ribosomal protein.a neurospora crassa cytoplasmic ribosomal protein gene, crp-5, has been isolated and characterized. the cdna was isolated by a differential screening of a cdna library for glucose-inducible genes. the cdna was subsequently used to identify and isolate crp-5 genomic sequences. computer analysis of the dna sequences showed that they contain an open reading frame which encodes a protein homologous to the rat ribosomal protein s26. the crp-5 mrna levels are regulated in a carbon-source-dependent man ...19938467530
deletion mutations in the speed operon: spermidine is not essential for the growth of escherichia coli.null mutants of escherichia coli were constructed that cannot synthesize spermidine, because of deletions in the gene encoding s-adenosylmethionine decarboxylase. these mutants are still able to grow at near normal rates in purified media deficient in polyamines. these results in e. coli differ from recent findings that null mutants of saccharomyces cerevisiae and of neurospora crassa have an absolute growth requirement for spermidine.19938472951
cloning and analysis of the mating type genes from cochliobolus heterostrophus.cochliobolus heterostrophus, a heterothallic ascomycete, has a single mating type locus with two alternate forms called mat-1 and mat-2. mat-1 was cloned by complementing a mat-2 strain using a cosmid library from a mat-1 strain and screening for a homothallic transformant. the cosmid recovered from this transformant was able to re-transform a mat-2 strain to homothallism and mat identity was proven by restriction fragment length polymorphism and conventional genetic mapping. all homothallic tra ...19938479433
cloning and sequencing of the genes coding for the a and b subunits of vacuolar-type na(+)-atpase from enterococcus hirae. coexistence of vacuolar- and f0f1-type atpases in one bacterial cell.the eubacterium enterococcus hirae atcc 9790 possesses a h(+)-translocating atpase, and the deduced amino acid sequences of the genes coding for this enzyme have indicated that it is a typical f0f1-type atpase (shibata, c., ehara, t., tomura, k., igarashi, k., and kobayashi, h. (1992) j. bacteriol. 174, 6117-6124). we cloned the ntpa and ntpb genes coding for the a and b subunits, respectively, of na(+)-translocating atpase from the same bacterium, and the full amino acid sequences of the two su ...19938505293
the beta subunit of the mitochondrial processing peptidase from rat liver: cloning and sequencing of a cdna and comparison with a proposed family of metallopeptidases.most nuclearly encoded mitochondrial proteins are synthesized with amino-terminal leader peptides that are removed by the mitochondrial processing peptidase (mpp) after translocation. earlier we reported cloning and sequencing of a cdna for the larger subunit (mpp alpha subunit) of this enzyme from rat liver mitochondria. we have now completed the cloning and sequencing of a cdna encoding the smaller subunit of the enzyme (mpp beta subunit) from the same source. the cdna consists of 1570 bp: 17 ...19938506385
developmental regulation of the gene for formate dehydrogenase in neurospora crassa.we have isolated and characterized a gene, fdh, from neurospora crassa which is developmentally regulated and which produces formate dehydrogenase activity when expressed in escherichia coli. the gene is closely linked (less than 0.6 kb apart) to the leu-5 gene encoding mitochondrial leucyl-trna synthetase; the two genes are transcribed convergently from opposite strands. the expression patterns of these genes differ: fdh mrna is found only during conidiation and early germination and is not det ...19938509325
pisatin resistance in dictyostelium discoideum and neurospora crassa: comparison of mutant phenotypes.the pea phytoalexin pisatin, at its inhibitory concentration, was shown to have two distinct inhibitory effects on amoebae of the cellular slime mould dictyostelium discoideum. one effect was cytolytic and was demonstrable even in non-growing cells whereas the second effect was observed only under conditions favourable to growth. pretreatment with a sublethal concentration of pisatin induced the amoebae to acquire resistance to both these effects. mutations in nysc that alter membrane sterols an ...19938126430
the role of the nad-dependent glutamate dehydrogenase in restoring growth on glucose of a saccharomyces cerevisiae phosphoglucose isomerase mutant.phosphoglucose isomerase pgi1-deletion mutants of saccharomyces cerevisiae cannot grow on glucose as the sole carbon source and are even inhibited by glucose. these growth defects could be suppressed by an over-expression on a multi-copy plasmid of the structural gene gdh2 coding for the nad-dependent glutamate dehydrogenase. gdh2 codes for a protein with 1092 amino acids which is located on chromosome xii and shows high sequence similarity to the neurospora crassa nad-glutamate dehydrogenase. s ...19937901008
the leu-1 gene of neurospora crassa: nucleotide and deduced amino acid sequence comparisons.the neurospora crassa leu-1 gene encodes beta-isopropylmalate dehydrogenase (ipmdh; ec 1.1.1.85), an enzyme in the leucine biosynthetic pathway. we determined the nucleotide sequence of the entire leu-1 gene and of four independent cdna clones. by comparing the genomic and cdna sequences, four introns were identified in the 5' portion of the gene and a single open reading frame was established. one of the introns is located within the 5'-noncoding region of the transcript. the deduced amino acid ...19938294021
genesis of eukaryotic transcriptional activator and repressor proteins by splitting a multidomain anabolic enzyme.the genes necessary for the correctly regulated catabolism of quinate in aspergillus nidulans and neurospora crassa are controlled at the level of transcription by a dna-binding activator protein and a repressor protein that directly interact with one another. the repressor protein is homologous throughout its length with the three c-terminal domains of a pentafunctional enzyme catalysing five consecutive steps in the related anabolic shikimate pathway. we now report that the activator protein i ...19938294040
the alkane-inducible candida maltosa ali1 gene product is an nadh:ubiquinone oxidoreductase subunit homologue.the ali1 gene product in candida maltosa was previously shown to be essential for n-alkane assimilation, possibly as a transcription factor [hwang et al., gene 106 (1991) 61-69]. we show that the predicted sequence is highly homologous to a subunit of respiratory complex i from another fungus, neurospora crassa, and from bos taurus. the predicted protein contains a motif conserved in this subunit from mitochondria, chloroplasts and bacteria. it also contains an n-terminal sequence that suggests ...19938299970
pclk1 and pclt5--two linear mitochondrial plasmids from unrelated claviceps purpurea strains: a comparison.pclt5, a linear mitochondrial (mt) plasmid from claviceps purpurea, strain t5, was sequenced and compared to pclk1, a linear mt plasmid from an unrelated c. purpurea strain. both plasmids have terminal proteins (tps) at their inverted terminal repeats (tlr). the tlrs of both plasmids show short conserved sequences, which are probably involved in plasmid transcription and replication. the coding capacity of pclt5 and pclk1 is similar: there are two large orfs (orf1 and orf2) homologous to the dna ...19938302935
ageing of neurospora crassa. vi. cytochemical and cytological correlates of senescence in three model systems. 1993204837
ageing of neurospora crassa. v. lipid peroxidation and decay of respiratory enzymes in an inositol auxotroph. 1993204836
molecular cloning of a cdna encoding calmodulin from neurospora crassa.a full-length cdna encoding neurospora crassa calmodulin was isolated from a lambda zap ii cdna expression library. the open reading frame encodes a protein of 148 amino acid residues with a calculated m(r) of 16,865 da. using site-directed mutagenesis, the complete cdna was ligated into a trc promoter-regulated bacterial expression vector to allow expression of n. crassa calmodulin in e. coli. the expressed protein was found to be identical to the native protein on the basis of some of its bioc ...19938467912
the 12.3 kda subunit of complex i (respiratory-chain nadh dehydrogenase) from neurospora crassa: cdna cloning and chromosomal mapping of the gene.the 12.3 kda subunit of complex i (respiratory-chain nadh dehydrogenase) is a nuclear-coded protein of the hydrophobic fragment of the enzyme. we have isolated and sequenced a full-length cdna clone coding for this polypeptide. the deduced protein is 104 amino acid residues long with a molecular mass of 12305 da. this particular subunit of complex i lacks a cleavable mitochondrial targeting sequence. in agreement with its localization within complex i, we have found that this subunit behaves lik ...19938098209
molecular characterization of conventional and new repeat-induced mutants of nit-3, the structural gene that encodes nitrate reductase in neurospora crassa.nitrate reductase of neurospora crassa is a dimeric protein composed of two identical subunits, each possessing three separate domains, with flavin, heme, and molybdenum-containing cofactors. a number of mutants of nit-3, the structural gene that encodes neurospora nitrate reductase, have been characterized at the molecular level. amber nonsense mutants of nit-3 were found to possess a truncated protein detected by a specific antibody, whereas ssu-1-suppressed nonsense mutants showed restoration ...19938479443
differential effects of anticytoskeletal compounds on the localization and chemical patterns of actin in germinating conidia of neurospora crassa.anti-actin drugs, cytochalasins a and b, inhibited both normal single, and benomyl-induced multiple, germ tube outgrowth from conidia of neurospora crassa. actin was cytochemically found to be concentrated in each of the benomyl-induced germ tube tips. no significant quantitative changes either in total actin or its isoforms were measured in the inhibitor-treated germlings. while intact microtubules are required for normal, monopolar axiation of the germ tube, they appear not to be necessary for ...19938472908
reactive oxygen species associated with cell differentiation in neurospora crassa.the conidiation process of neurospora crassa is characterized by three morphogenetic events: adhesion of hyphae, development of aerial hyphae, and conidia formation. at the onset of all three events a spontaneous, low-level chemiluminescence was detected, indicating the formation of reactive oxygen species. hyperoxic conditions increased chemiluminescence and accelerated differentiation. hypoxic conditions abolished both chemiluminescence and differentiation. chemiluminescence was enhanced by lu ...19938458586
plasmid diversity in senescent and nonsenescent strains of neurospora.a sample of 171 natural isolates of neurospora crassa and neurospora intermedia was tested for senescence. of these, 28 strains senesced within the duration of the experiment. these senescent strains, together with a selection of nonsenescent strains, were examined for the presence of mitochondrial plasmids. this was done by digesting mitochondrial dna preparations with proteinase k, and running these samples on agarose gels. most of the strains examined, both senescent and nonsenescent, contain ...19938455555
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