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sequence similarities within the family of dihydrolipoamide acyltransferases and discovery of a previously unidentified fungal enzyme.a composite protein sequence database was searched for amino acid sequences similar to the c-terminal domain of the dihydrolipoamide acetyltransferase subunit (e2p) of the pyruvate dehydrogenase complex of escherichia coli. nine sequences with extensive similarity were found, of which eight were e2 subunits. the other was for a putative mitochondrial ribosomal protein, mrp3, from neurospora crassa. alignment of the mrp3 and e2 sequences showed that the similarity extends through the entire mrp3 ...19911825611
the neurospora crassa carotenoid biosynthetic gene (albino 3) reveals highly conserved regions among prenyltransferases.in the filamentous fungus neurospora crassa the biosynthesis of carotenoids is regulated by blue light. here we report the characterization of the albino-3 (al-3) gene of n. crassa, which encodes the carotenoid biosynthetic enzyme geranylgeranyl-pyrophosphate synthetase. this is the first geranylgeranyl-pyrophosphate synthetase gene isolated. nucleotide sequence comparison of al-3 genomic and cdna clones revealed that the al-3 gene is not interrupted by introns. transcription of the al-3 gene ha ...19911826006
identification of the active-site lysine residues of two biosynthetic 3-dehydroquinases.the lysine residues involved in schiff-base formation at the active sites of both the 3-dehydroquinase component of the pentafunctional arom enzyme of neurospora crassa and of the monofunctional 3-dehydroquinase of escherichia coli were labelled by treatment with 3-dehydroquinate in the presence of nab3h4. radioactive peptides were isolated by h.p.l.c. following digestion with cnbr (and in one case after further digestion with trypsin). the sequence established for the n. crassa peptide was alqh ...19911826831
all internal promoter elements of neurospora crassa 5 s rrna and trna genes, including the a boxes, are functionally gene-specific.the internal control elements of neurospora crassa 5 s genes include an a box and a c box as in xenopus and saccharomyces cerevisiae, plus a novel element, the ribo box at position +18 to +34. the ribo box is also found in the 40 s rrna promoter and a ribosomal protein gene but is absent from trna genes in n. crassa. the 5 s a box diverges from the trna a box consensus at two positions. we tested whether replacement of the 5 s a box with a trnaleu a box sequence would increase 5 s gene transcrip ...19911827115
sulfate transport in neurospora crassa: regulation, turnover, and cellular localization of the cys-14 protein.uptake of inorganic sulfate in neurospora crassa is governed by the sulfur regulatory circuit and is under the control of positively and negatively acting regulatory genes. two genetically and biochemically distinct systems are responsible for the uptake of sulfate from the environment. one of these, sulfate permease ii, encoded by the cys-14 gene, functions primarily in mycelia. a defined region of the cys-14 protein was highly expressed in escherichia coli and purified. anti-cys-14 antibody wa ...19911827594
duplication of leader sequence for protein targeting to mitochondria leads to increased import efficiency.we describe a novel method for enhancing protein import into mitochondria, by tandemly duplicating the n-terminal cleavable leader peptide using a gene manipulation strategy. the import into isolated yeast mitochondria of passenger proteins (yeast mitochondrial atp synthase subunits 8 and 9 and some mutagenised derivatives) that show little or no import when endowed with one such leader (that of neurospora crassa mitochondrial atp synthase subunit 9) is remarkably improved when the leader is tan ...19911828039
heterologous expression and regulation of the neurospora crassa nit-4 pathway-specific regulatory gene for nitrate assimilation in aspergillus nidulans.the nira gene of aspergillus nidulans and the nit-4 gene of neurospora crassa appear to be equivalent pathway-specific regulatory genes which mediate nitrate induction of nitrate reductase and nitrite reductase (nr and nir) activities. we have transformed the nit-4 wild-type (wt) gene into the a. nidulans loss-of-function (pleiotropic negative) nira 1 mutant strain. the nit-4 gene was found to complement the nira 1 mutation, thus permitting the nira 1 mutant strain to grow on nitrate or nitrite ...19911829047
one hour in 1 ata oxygen enhances rat alveolar macrophage chemiluminescence and fungal cytotoxicity.the purpose of this study was to determine if 100% o2 would enhance rat pulmonary alveolar macrophage (pam) oxidative killing of conidia of the fungus neurospora crassa. first, we found that incubation in 100% o2 had no effect on conidia viability in the absence of pam. we obtained resident pam from nonpretreated anesthetized male sprague-dawley rats by bronchoalveolar lavage. compared with similar air exposures we found that 1 h in vitro exposure of pam to 100% o2 (1.0 atmosphere absolute) incr ...19911829328
suppression of the cr-1 mutation in neurospora crassa.we have cloned a dna fragment, which hybridized with the adenylate cyclase gene (cyr1) of saccharomyces cerevisiae, from genomic dna libraries of neurospora crassa. the cr-1 mutation was able to be suppressed by introducing this dna fragment on a cosmid vector, judging from recovery of the adenylate cyclase activity and the abnormal morphology.19911829616
the beta-oxidation system in catalase-free microbodies of the filamentous fungus neurospora crassa. purification of a multifunctional protein possessing 2-enoyl-coa hydratase, l-3-hydroxyacyl-coa dehydrogenase, and 3-hydroxyacyl-coa epimerase activities.a trifunctional beta-oxidation protein, designated tfp, was purified to apparent homogeneity from oleate-induced mycelia of neurospora crassa. 2-enoyl-coa hydratase, l-3-hydroxyacyl-coa dehydrogenase, and 3-hydroxyacyl-coa epimerase activities copurified in constant ratios with this protein when crude extracts were subjected to cation-exchange, dye-ligand, and adsorption chromatography. trifunctionality was substantiated by coinciding enzyme activity ratios during the last two purification steps ...19911830048
the neurospora crassa cyt-20 gene encodes cytosolic and mitochondrial valyl-trna synthetases and may have a second function in addition to protein synthesis.the cyt-20-1 mutant of neurospora crassa is a temperature-sensitive, cytochrome b- and aa3-deficient strain that is severely deficient in both mitochondrial and cytosolic protein synthesis (r.a. collins, h. bertrand, r.j. lapolla, and a.m. lambowitz, mol. gen. genet. 177:73-84, 1979). we cloned the cyt-20+ gene by complementation of the cyt-20-1 mutation and found that it contains a 1,093-amino-acid open reading frame (orf) that encodes both the cytosolic and mitochondrial valyl-trna synthetases ...19911830127
nadh:ubiquinone oxidoreductase from bovine mitochondria. cdna sequence of a 19 kda cysteine-rich subunit.the sequence of a 19 kda subunit of nadh:ubiquinone oxidoreductase (complex i) from bovine heart mitochondria has been determined by a new strategy based on the polymerase chain reaction. the subunits of the enzyme were resolved in a polyacrylamide gel by two-dimensional isoelectric focusing and electrophoresis under denaturing conditions, transferred to a poly(vinylidene difluoride) membrane, and the n-terminal sequence was determined on the stained 19 kda protein up to residue 27. this informa ...19911830204
short dispersed repeats localized in spacer regions of chlamydomonas reinhardtii mitochondrial dna.in the mtdna of chlamydomonas reinhardtii, a unicellular green alga, we have identified a set of short repeated sequences up to 65 nucleotides long, each of which contains the palindromic consensus motif ctcgg(n4-14)ccgag. most of these repeated elements are localized in spacer regions that flank the transcribed coding regions of c. reinhardtii mtdna. these algal mitochondrial repeats have features reminiscent of short repeats in some fungal mtdnas, such as gc clusters in saccharomyces cerevisia ...19911831072
2-amino-n6-hydroxyadenine induces gene/point mutations and multiple-locus mutations, but not multilocus deletion mutations, in the ad-3 region of a two-component heterokaryon of neurospora crassa.the mutagenicity of 2-amino-n6-hydroxyadenine (aha) has been studied in neurospora crassa by treating a two-component heterokaryon (h-12) and recovering specific-locus mutations induced in the ad-3 region. this assay system permits the identification of ad-3a and/or ad-3b mutants resulting from gene/point mutations, multilocus deletion mutations, and multiple-locus mutations of various genotypes, involving one or both loci. genetic characterization of the ad-3 mutants recovered from experiments ...19911831243
cloning the mating types of the heterothallic fungus podospora anserina: developmental features of haploid transformants carrying both mating types.dnas that encode the mating-type functions (mat+ and mat-) of the filamentous fungus podospora anserina were cloned with the use of the mating-type a probe from neurospora crassa. cloning the full mat information was ascertained through gene replacement experiments. molecular and functional analyses of haploid transformants carrying both mating types lead to several striking conclusions. mat+ mat- strains are dual maters. however, the resident mat information is dominant to the mat information a ...19911831427
evolutionary conservation of a microbody targeting signal that targets proteins to peroxisomes, glyoxysomes, and glycosomes.peroxisomes, glyoxysomes, glycosomes, and hydrogenosomes have each been classified as microbodies, i.e., subcellular organelles with an electron-dense matrix that is bound by a single membrane. we investigated whether these organelles might share a common evolutionary origin by asking if targeting signals used for translocation of proteins into these microbodies are related. a peroxisomal targeting signal (pts) consisting of the cooh-terminal tripeptide serine-lysine-leucine-cooh has been identi ...19911831458
mutational analysis of the dna-binding domain of the cys3 regulatory protein of neurospora crassa.cys-3, the major sulfur regulatory gene of neurospora crassa, activates the expression of a set of unlinked structural genes which encode sulfur catabolic-related enzymes during conditions of sulfur limitation. the cys-3 gene encodes a regulatory protein of 236 amino acid residues with a leucine zipper and an upstream basic region (the b-zip region) which together may constitute a dna-binding domain. the b-zip region was expressed in escherichia coli to examine its dna-binding activity. the b-zi ...19911831537
primary structures of two subunits of nadh: ubiquinone reductase from neurospora crassa concerned with nadh-oxidation. relationship to a soluble nad-reducing hydrogenase of alcaligenes eutrophus.the primary structures of the nuclear-encoded 51 kda and 78 kda subunits of the respiratory chain nadh: ubiquinone reductase (complex i) from neurospora crassa mitochondria were determined by sequencing cdna and the n-terminus of the mature proteins. both subunits are related to the soluble nad-reducing hydrogenase of the bacterium alcaligenes eutrophus. sequence comparison between these subunits, the corresponding subunits of the bovine complex i and the bacterial nad-reducing hydrogenase furth ...19911832016
molecular organisation of the malate synthase genes of aspergillus nidulans and neurospora crassa.the sequencing and comparison of the genes encoding the glyoxylate bypass enzyme malate synthase of aspergillus nidulans (acue) and neurospora crassa (acu-9) are presented. the predicted amino acid sequences of the a. nidulans and n. crassa enzymes are 538 and 542 residues respectively and the proteins are 87% homologous. in fungi, the malate synthase proteins are located in glyoxysomes and the deduced acue and acu-9 proteins both contain a c-terminal s-k-l sequence, which has been implicated in ...19911832736
nadh:ubiquinone oxidoreductase from bovine heart mitochondria. cdna sequences of the import precursors of the nuclear-encoded 39 kda and 42 kda subunits.the 39 kda and 42 kda subunits of nadh:ubiquinone oxidoreductase from bovine heart mitochondria are nuclear-coded components of the hydrophobic protein fraction of the enzyme. their amino acid sequences have been deduced from the sequences of overlapping cdna clones. these clones were amplified from total bovine heart cdna by means of the polymerase chain reaction, with the use of complex mixtures of oligonucleotide primers based upon fragments of protein sequence determined at the n-terminals o ...19911832859
the initiation site for recombination cog is at the 3' end of the his-3 gene in neurospora crassa.recombination at his-3 in neurospora crassa is thought to be initiated through a site designated cog which lies in the his-3 to ad-3 interval of linkage group i. fragments of the his-3 gene were used to transform various his-3 mutant alleles to prototrophy in order to link the genetic map to the nucleotide sequence. it was established that cog is at the 3' end of his-3 and is therefore not the his-3 promoter. this suggests that cog may be dissimilar to a number of yeast recombinators which are a ...19911833619
antibodies against the 59 kda polypeptide of the n. crassa 8-10 nm filaments immunodetect a 59 kda polypeptide in specialized rat epithelial cells.p59nc is a polypeptide associated with bundles of cytoplasmic and nuclear filamentous structures of 8-10 nm of diameter in neurospora crassa cells. it is immunologically unrelated to both higher and lower eucaryotic tubulin and actin proteins and is detected weakly by the anti ifa monoclonal antibody. we analyze here the immunological relationship between p59nc and intermediate filament (if) mammalian proteins by using anti p59nc, anti keratin, anti vimentin and anti ifa antibodies. anti p59nc a ...19911833626
cloning and functional characterization of a eucaryotic dna photolyase gene from neurospora crassa.we cloned a genomic fragment of a photolyase gene from neurospora crassa by polymerase chain reaction using synthesized oligonucleotide primers designed from the most conserved amino acid sequences among photolyases of various organisms. using the cloned fragment as a hybridization probe we isolated a genomic fragment and cdna clones encoding the complete photolyase gene of this organism. the amino acid sequence of the photolyase deduced from the determined nucleotide sequence indicates a protei ...19911833725
[synthesis and antifungal activity of chlorobenzyl benzylidene thiazolidinediones and substituted of imidazolidinediones].the synthesis of six benzylidene thiazolidinediones and four benzylidene imidazolidinediones is described. in order to investigate their antifungal activity, they are evaluated against microorganism such as candida albicans, neurospora crassa, staphylococcus aureus and escherichia coli.19911834006
sequence of the nuclear atp synthase subunit 9 gene of podospora anserina: lack of similarity to the mitochondrial genome.the nuclear gene coding for the mitochondrial subunit 9 of the f0f1-atp synthase complex was isolated from a genomic library of podospora anserina. nucleotide sequencing revealed an open reading frame capable to code for 144 amino acids including an amino-terminal pre-sequence of 63 amino acid residues for mitochondrial import of the pre-proteolipid. the p. anserina proteolipid shows extensive sequence identity with the corresponding gene products of the related filamentous fungi neurospora cras ...19911834355
expression of meiotic drive elements spore killer-2 and spore killer-3 in asci of neurospora tetrasperma.it was shown previously that when a chromosomal spore killer factor is heterozygous in neurospora species with eight-spored asci, the four sensitive ascospores in each ascus die and the four survivors are all killers. sk-2k and sk-3k are nonrecombining haplotypes that segregate with the centromere of linkage group iii. no killing occurs when either one of these killers is homozygous, but each is sensitive to killing by the other in crosses of sk-2k x sk-3k. in the present study, sk-2k and sk-3k ...19911834522
utilization of the specific-locus assay in the ad-3 region of two-component heterokaryons of neurospora for risk assessment of environmental chemicals.the utilization of the specific-locus assay in the ad-3 region of two-component heterokaryons of neurospora crassa is compared with that of other eukaryotic assay systems for the evaluation of the mutagenic effects of environmental chemicals. in contrast to other in vitro specific-locus assays, the neurospora assay can detect mutations not only at the ad-3a and ad-3b loci but also recessive lethal mutations elsewhere in the genome. mutational damage in this system can be characterized readily by ...19911834935
qualitative differences in the spectra of genetic damage in 2-aminopurine-induced ad-3 mutants between nucleotide excision-repair-proficient and -deficient strains of neurospora crassa.the mutagenic effects of 2-aminopurine (2ap) have been compared in the adenine-3 (ad-3) region of two-component heterokaryons of neurospora crassa: nucleotide excision repair-proficient (uvs-2+/uvs-2+) heterokaryon 12 (h-12) and nucleotide excision repair-deficient (uvs-2/uvs-2) heterokaryon 59 (h-59). this forward-mutation, morphological and biochemical, specific-locus assay system permits the recovery of ad-3a and/or ad-3b mutants in 3 major classes: gene/point mutations, multilocus deletion m ...19911834936
characterization of two beta-tubulin genes from geotrichum candidum.the beta-tubulin genes g beta 1 and g beta 2 from the phytopathogenic hemiascomycete geotrichum candidum were found to be highly diverged in amino acid sequence from those of other filamentous fungi. g beta 1 and g beta 2 were also divergent from each other, with the coding regions sharing only 66% nucleotide sequence homology and 64% amino acid identity. however, the proteins shared 82% similarity and only 25 of the 161 non-identical amino acid substitutions were non-conservative. the organizat ...19911836049
chitin synthase 1 plays a major role in cell wall biogenesis in neurospora crassa.in filamentous fungi, chitin is a structural component of morphologically distinct structures assembled during various phases of growth and development. to investigate the role of chitin synthase in cell wall biogenesis in neurospora crassa, we cloned a chitin synthase structural gene and examined the consequences of its inactivation. using degenerate oligonucleotide mixtures designed on the basis of conserved sequences of the saccharomyces cerevisiae chs1 and chs2 polypeptides, a dna fragment e ...19911836444
coordinate expression of ribosomal protein genes in neurospora crassa and identification of conserved upstream sequences.the relative levels of rrnas and ribosomal proteins are coordinately regulated by growth rate and carbon nutrition in neurospora crassa. however, little is known about the mechanisms involved. to investigate the transcriptional regulation of ribosomal protein genes in n. crassa, we cloned and sequenced a ribosomal protein gene (crp-3). the inferred crp-3 protein sequence shares 89% and 83% homology at its n-terminus with the yeast rp51 and the human s17 ribosomal proteins respectively. the crp-3 ...19911836561
molecular cloning, characterization and analysis of the regulation of the aro2 gene, encoding chorismate synthase, of saccharomyces cerevisiae.we describe here the cloning, characterization and analysis of the regulation of the aro2 gene of saccharomyces cerevisiae, the first reported study of a eukaryotic gene encoding chorismate synthase (e.c. 4.6.1.4). the gene contains an orf of 1128 bp, encoding a protein with a calculated molecular mass of 40.8 kda. aro2 is regulated under the 'general control system' for amino acid biosynthesis by the transcriptional activator gcn4 which binds in vitro at three sites within the aro2 promoter. th ...19911837329
an electroporation-based system for high-efficiency transformation of germinated conidia of filamentous fungi.a rapid and efficient electroporation procedure has been developed for transformation of germinating conidia of filamentous fungi. pretreatment of conidial preparations with a cell wall weakening agent, such as beta-glucuronidase, was found to be essential for successful transformation. using the qa-2+ gene of neurospora crassa, encoding the catabolic dehydroquinase, as a selectable marker with a double-mutant host strain, auxotrophic for aromatic amino acids, integration of the plasmid was obse ...19911838030
cloning of a saccharomyces cerevisiae gene encoding a protein homologous to allantoicase of neurospora crassa. 19911839481
nit-4, a pathway-specific regulatory gene of neurospora crassa, encodes a protein with a putative binuclear zinc dna-binding domain.nit-4, a pathway-specific regulatory gene in the nitrogen circuit of neurospora crassa, is required for the expression of nit-3 and nit-6, the structural genes which encode nitrate and nitrite reductase, respectively. the complete nucleotide sequence of the nit-4 gene has been determined. the predicted nit4 protein contains 1,090 amino acids and appears to possess a single zn(ii)2cys6 binuclear-type zinc finger, which may mediate dna binding. site-directed mutagenesis studies demonstrated that c ...19911840634
nucleotide and derived amino acid sequences of the major porin of comamonas acidovorans and comparison of porin primary structures.the dna sequence of the gene which codes for the major outer membrane porin (omp32) of comamonas acidovorans has been determined. the structural gene encodes a precursor consisting of 351 amino acid residues with a signal peptide of 19 amino acid residues. comparisons with amino acid sequences of outer membrane proteins and porins from several other members of the class proteobacteria and of the chlamydia trachomatis porin and the neurospora crassa mitochondrial porin revealed a motif of eight r ...19911848840
the iron-sulfur clusters in the two related forms of mitochondrial nadh: ubiquinone oxidoreductase made by neurospora crassa.two related forms of the respiratory-chain complex, nadh: ubiquinone oxidoreductase (complex i) are synthesized in the mitochondria of neurospora crassa. normally growing cells make a large, piericidin-a-sensitive form, which consists of some 23 different nuclear- and 6-7 mitochondrially encoded subunits. cells grown in the presence of chloramphenicol make a small, piericidin-a-insensitive form which consists of only approximately 13 nuclear-encoded subunits. the subunits of the small form are e ...19911849820
cysteinyl-trna synthetase is a direct descendant of the first aminoacyl-trna synthetase.the gene encoding the cysteinyl-trna synthetase of e. coli was cloned from an e. coli genomic library made in lambda 2761, a lambda vector which can integrate and which carries a chloramphenicol resistance gene. a thermosensitive cyss mutant of e. coli was lysogenised and chloramphenicol-resistant colonies able to grow at 42 degrees c were selected to isolate phages containing the wild-type cyss gene. the sequence of the gene was determined. it codes for a 461 amino-acid protein and includes the ...19911864365
phylogenetic analysis of five medically important candida species as deduced on the basis of small ribosomal subunit rna sequences.the classification of species belonging to the genus candida berkhout is problematic. therefore, we have determined the small ribosomal subunit rna (srrna) sequences of the type strains of three human pathogenic candida species; candida krusei, c. lusitaniae and c. tropicalis. the srrna sequences were aligned with published eukaryotic srrna sequences and evolutionary trees were inferred using a matrix optimization method. an evolutionary tree comprising all available eukaryotic srrna sequences, ...19911865186
cloning, sequencing and expression of the schwanniomyces occidentalis nadp-dependent glutamate dehydrogenase gene.the cloned nadp-specific glutamate dehydrogenase (gdh) genes of aspergillus nidulans (gdha) and neurospora crassa (am) have been shown to hybridize under reduced stringency conditions to genomic sequences of the yeast schwanniomyces occidentalis. using 5' and 3' gene-specific probes, a unique 5.1 kb bcli restriction fragment that encompasses the entire schwanniomyces sequence has been identified. a recombinant clone bearing the unique bcli fragment has been isolated from a pool of enriched clone ...19911934128
analysis of the altered mrna stability (ams) gene from escherichia coli. nucleotide sequence, transcriptional analysis, and homology of its product to mrp3, a mitochondrial ribosomal protein from neurospora crassa.the product of the altered mrna stability (ams) gene of escherichia coli is involved in decay of mrna. the complete nucleotide sequence of a 4-kilobase bamhi restriction fragment containing the ams coding sequence was determined. transcription of the ams gene was analyzed by high resolution s1 mapping. a promoter was found with a homology score of 58% 361 nucleotides upstream from the start codon of ams. the ams structural gene consists of an open reading frame of 2,445 nucleotides. the protein ...19911704367
voltage gating of the mitochondrial outer membrane channel vdac is regulated by a very conserved protein.soluble protein preparations obtained from the mitochondrial fractions of three very different organisms, neurospora crassa, rat, and potato, were discovered to greatly enhance the voltage sensitivity of the mitochondrial outer membrane channel, vdac. the active ingredient, referred to as the vdac modulator, increased the rate of voltage-dependent channel closure by approximately 10-fold. the modulator from one species increased the closing rate of vdac channels from all three species. the activ ...19911705100
localization of actin and characterization of its isoforms in the hyphae of neurospora crassa.the actin of neurospora crassa wild type strain st. lawrence has been purified, characterized and localized. a fungal 43 kda protein was isolated by affinity chromatography on dnase i-sepharose. this protein was identified as actin on immunoblots when an anti-actin monoclonal antibody raised against chicken gizzards was used as a probe. after two-dimensional gel electrophoresis three actin isoforms were detected. the distribution of actin in hyphae was examined by fitc-phalloidin staining of for ...19911706289
incipient mitochondrial evolution in yeasts. ii. the complete sequence of the gene coding for cytochrome b in saccharomyces douglasii reveals the presence of both new and conserved introns and discloses major differences in the fixation of mutations in evolution.we have determined the complete sequence of the mitochondrial gene coding for cytochrome b in saccharomyces douglasii. the gene is 6310 base-pairs long and is interrupted by four introns. the first one (1311 base-pairs) belongs to the group id of secondary structure, contains a fragment open reading frame with a characteristic giy ... yig motif, is absent from saccharomyces cerevisiae and is inserted in the same site in which introns 1 and 2 are inserted in neurospora crassa and podospora anseri ...19911708831
core i protein of bovine ubiquinol-cytochrome-c reductase; an additional member of the mitochondrial-protein-processing family. cloning of bovine core i and core ii cdnas and primary structure of the proteins.core i and core ii proteins are the largest nuclear-encoded subunits of the mitochondrial ubiquinol-cytochrome-c reductase (bc1 complex) lacking redox prosthetic groups. cdna clones of the two bovine core proteins have been isolated by the screening of lambda zap cdna libraries either with an oligonucleotide probe based on the sequence of an internal peptide or with a polymerase-chain-reaction-amplified fragment. the core i precursor protein consists of 362 amino acids with a 34-amino-acid prese ...19911712295
insulin-induced stimulation of protein phosphorylation in neurospora crassa cells.1) insulin stimulated the phosphorylation of at least 14 discrete proteins in neurospora crassa cells. specific proteins were phosphorylated at serine, threonine, and tyrosine residues, as determined by phosphoamino acid analysis of discrete spots on two-dimensional gels. 2) insulin stimulated the phosphorylation by [gamma-32p]atp of at least six discrete proteins in solubilized n. crassa membrane preparations at serine and tyrosine residues. 3) a phosphotyrosine-containing protein of 38 kda, pi ...19911717334
a nuclear gene with many introns encoding ammonium-inducible chloroplastic nadp-specific glutamate dehydrogenase(s) in chlorella sorokiniana.chlorella sorokiniana possesses ammonium-inducible, chloroplastic, nadp-specific glutamate dehydrogenase (nadp-gdh) homo- or heterohexamers composed of alpha- and/or beta-subunits which were previously shown to derive from precursor protein(s) of identical size. from the present studies, data are consistent with these two subunits being encoded by a single nuclear gene. the nadp-gdh gene is greater than 7 kb in length due to the presence of at least 21 introns, an unusually large number for a eu ...19911718478
oxidation of molybdopterin in sulfite oxidase by ferricyanide. effect on electron transfer activities.the attenuation of the sulfite:cytochrome c activity of sulfite oxidase upon treatment with ferricyanide was demonstrated to be the result of oxidation of the pterin ring of the molybdenum cofactor in the enzyme. oxidation of molybdopterin (mpt) was detected in several ways. ferricyanide treatment not only abolished the ability of sulfite oxidase to serve as a source of mpt to reconstitute the aponitrate reductase in extracts of the neurospora crassa mutant nit-1 but also eliminated the ability ...19912002036
identification of molybdopterin guanine dinucleotide in formate dehydrogenase from methanobacterium formicicum.the pterin cofactor in formate dehydrogenase isolated from methanobacterium formicium is identified as molybdopterin guanine dinucleotide. the pterin, stabilized as the alkylated, dicarboxamidomethyl derivative, is shown to have absorption and chromatographic properties identical to those of the previously characterized authentic compound. treatment with nucleotide pyrophosphatase produced the expected degradation products gmp and carboxyamidomethyl molybdopterin. the molybdopterin guanine dinuc ...19912037231
integrative transformation of the ascomycete podospora anserina: identification of the mating-type locus on chromosome vii of electrophoretically separated chromosomes.protoplasts of wild-type strain s and a long-lived extrachromosomal mutant (al2) of the ascomycete podospora anserina were transformed using a plasmid (pan7-1) which contains the hygromycin b phosphotransferase gene (hph) of escherichia coli under the control of aspergillus nidulans regulatory sequences. after optimizing the transformation procedure, transformation efficiencies of 15-21 transformants/micrograms plasmid dna were obtained. using a second selectable vector (pbt3), which contains th ...19911367277
purification and characterization of chorismate synthase from euglena gracilis: comparison with chorismate synthases of plant and microbial origin.chorismate synthase was purified 1200-fold from euglena gracilis. the molecular mass of the native enzyme is in the range of 110 to 138 kilodaltons as judged by gel filtration. the molecular mass of the subunit was determined to be 41.7 kilodaltons by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. purified chorismate synthase is associated with an nadph-dependent flavin mononucleotide reductase that provides in vivo the reduced flavin necessary for catalytic activity. in vitro, flavi ...199116668543
blue light induction of conidiation-specific genes in neurospora crassa.the con genes of neurospora crassa are preferentially expressed during a developmental process known as conidiation. we present evidence indicating that transcription of con-5 and con-10 is also stimulated by blue light. transcription of these genes was not photoinducible in wc-1 and wc-2 mutant strains. the response of con-5 and con-10 to blue light was similar to that of al-1 and al-2, genes involved in carotenoid biosynthesis, and bli-3 and bli-4, blue light inducible genes.19911837079
adp-glucose transport by the chloroplast adenylate translocator is linked to starch biosynthesis.in organello starch biosynthesis was studied using intact chloroplasts isolated from spinach leaves (spinacia oleracea). immunoblot analysis using a specific antiserum against the mitochondrial adenylate (adp/atp) translocator of neurospora crassa shows the presence of an adenylate translocator protein in the chloroplast envelope membranes, similar to that existing in mitochondria and amyloplasts from cultured cells of sycamore (acer pseudoplatanus). the double silicone oil layer-filtering centr ...199116668585
a preproinsulin-like pseudogene from neurospora crassa.a segment of dna was amplified from the neurospora crassa genome by the polymerase chain reaction using several oligonucleotides coding for highly conserved domains in proinsulin as primers and probe. a genomic clone corresponding to this segment was isolated and the nucleotide sequence was determined. the deduced amino acid sequence of a part of this segment bears remarkable resemblance to preproinsulin, but lacks several requirements for transcription or translation and must therefore be consi ...19911838993
efficient dna pairing in a neurospora mutant defective in chromosome pairing.a neurospora crassa mutation, mei-2, affecting recombination and pairing of homologous chromosomes during meiosis, was characterized for its effect on repeat-induced point mutation (rip). we found that rip, which depends on recognition of dna sequence homology, is not inhibited by mei-2, suggesting that the defect in chromosome pairing of this mutant is not due to a defect in dna pairing and that dna pairing is not dependent on chromosome pairing.19911836526
mei-2, a mutagen-sensitive mutant of neurospora defective in chromosome pairing and meiotic recombination.a neurospora crassa mutation, mei-2, affecting meiosis and mutagen sensitivity, was characterized for its effect on meiotic recombination and chromosome pairing. results from homozygous mei-2 crosses involving distant markers on the same chromosome demonstrated a drastic reduction in meiotic recombination. however, mitotic recombination continued to occur. cytological observations indicated that pairing of homologous chromosomes in zygotene was greatly reduced or absent, resulting in aberrant se ...19911836525
regulation of laccase biosynthesis in the plant-pathogenic fungus cryphonectria parasitica by double-stranded rna.transmissible hypovirulence of the chestnut blight fungus, cryphonectria parasitica, is associated with cytoplasmic double-stranded-rna (dsrna) viruses. the fungal laccase has attracted interest because its activity is reduced in hypovirulent dsrna-containing strains. a laccase cdna clone was isolated by screening a cdna expression library with antibodies against the purified extracellular laccase. the amino acid sequence deduced from part of the cdna clone revealed high homology to other fungal ...19911744058
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
identification of a sterol mutant of neurospora crassa deficient in delta 14,15-reductase activity.a mutant (erg-3) of neurospora crassa resistant to the polyene antibiotic nystatin was compared with its sensitive, wild-type parent to detect differences in sterol composition using gas chromatography-mass spectrometry. the major sterol in wild-type mycelia, comprising 80% of the total, was ergosterol. the major sterols in mutant mycelia, comprising 86% of the total, were delta 8,14-sterols. it is proposed that the nystatin-resistant strain is unable to synthesize ergosterol because it lacks de ...19911838392
photoregulation of the albino-3 gene in neurospora crassa.in this paper we describe the light-regulated expression of albino-3 (al-3), a carotenoid biosynthetic gene of neurospora crassa, in the wild-type strain. our results suggest that the al-3 gene expression is regulated by the transcriptional activation of the gene and the low stability of its mrna. the activation of the al-3 gene does not require protein synthesis to occur. the kinetic analysis of the al-3 mrna reveals that the gene is transiently expressed even in continuous light, suggesting th ...19911837560
developmental expression of genes involved in conidiation and amino acid biosynthesis in neurospora crassa.the levels of transcripts for neurospora crassa genes concerned with cellular and metabolic functions changed dramatically at different stages of asexual development. transcripts for some conidiation-related (con) genes were present at high levels in conidiating cultures and in dormant conidia, but were absent or reduced during mycelial growth. levels of some con transcripts increased transiently during conidial germination, while others disappeared. transcripts for amino acid biosynthetic enzym ...19911834495
the induction and repair of (6-4) photoproducts in neurospora crassa.the (6-4) photoproduct lesion found in dna after uv irradiation is repaired by germinating neurospora crassa conidia. wild-type neurospora removes 80% of the (6-4) photoproduct in approximately 20 min and maximal repair is accomplished by 30 min with approximately 89% of the original lesions removed. mutagen-sensitive neurospora mutants belonging to the established excision repair epistasis group, uvs-2, are not defective in the removal of cyclobutane pyrimidine dimers. furthermore, we find thes ...19911719392
an apparent rare-codon effect on the rate of translation of a neurospora gene.as the result of two mutually compensating frameshift mutations, three successive codons with third-position a were generated in the neurospora crassa am (nadp-specific glutamate dehydrogenase: gdh) gene. these codons do not occur at all elsewhere in the gene and only infrequently in other highly expressed neurospora genes. the double-frameshift strain produces only 25 to 35% of the normal level of gdh, whether measured as enzyme activity or as immunoprecipitable protein, but its level of gdh mr ...19911834852
electron microscopic analysis of the peripheral and membrane parts of mitochondrial nadh dehydrogenase (complex i).two related forms of the respiratory chain nadh dehydrogenase (nadh:ubiquinone reductase or complex i) are synthesized in the mitochondria of neurospora crassa. normally growing cells make a large form that consists of 25 subunits encoded by nuclear dna and six to seven subunits encoded by mitochondrial dna. cells grown in the presence of chloramphenicol, however, make a smaller form comprising only 13 subunits, all encoded by nuclear dna. when the large enzyme is dissected by chaotropic agents ...19911834851
stability of chitin synthetase in cell-free preparations of a wild-type strain and a 'slime' variant of neurospora crassa.chitin synthetase activity in cell-free preparations from a wild-type strain and a 'slime' variant of neurospora crassa was monitored over many days in samples stored at 0 degrees c. total activity in whole-cell-free extracts and low-speed supernatants from both organisms was very unstable, losing more than 90% of the initial activity on storage at 0 degrees c for 96 h. chitin synthetase detection was not masked by chitinase activity present in the preparations. gel-filtration chromatography of ...19911838089
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
microcycle conidiation and its genetic basis in neurospora crassa.some wild isolates of neurospora show microcycle conidiation in liquid culture under continuous agitation. macroconidia from agar-grown mycelial cultures germinated in liquid and the germlings spontaneously produced conidia with no intervening mycelial phase. three types of microcycle conidiation were seen among progeny of n. crassa vickramam a x n. crassa a wild-type: (1) multinucleate blastoconidia produced by apical budding and septation, (2) multinucleate arthroconidia produced by holothalli ...19911836224
generation of new mutants of nmr, the negative-acting nitrogen regulatory gene of neurospora crassa, by repeat induced mutation.the repeat induced point mutation (rip) phenomenon has been used to generate new mutants of nmr, the negative nitrogen regulatory gene in neurospora crassa. the wild-type nmr gene was cotransformed along with the hygromycin b resistance gene into wild-type cells by selecting for hygromycin b resistance. following purification of primary transformants using microconidia, many chlorate-sensitive progeny were obtained from crosses to wild-type. detailed analyses of some of the progeny revealed that ...19911834354
the acyl-carrier protein in neurospora crassa mitochondria is a subunit of nadh:ubiquinone reductase (complex i).we determined the primary structure of a 9.6-kda subunit of the respiratory chain nadh:ubiquinone reductase (complex i) from neurospora crassa mitochondria and found a close relationship between this subunit and the bacterial or chloroplast acyl-carrier protein. the degree of sequence identity amounts to 80% in a region of 19 residues around the serine to which the phosphopantetheine is bound. the n-terminal presequence of the subunit has the characteristic features of a mitochondrial import seq ...19911832379
identification of the membrane-embedded regions of the neurospora crassa plasma membrane h(+)-atpase.reconstituted proteoliposomes containing functional neurospora crassa plasma membrane h(+)-atpase molecules oriented predominantly with their cytoplasmic surface exposed were treated with trypsin and then subjected to sepharose cl-6b column chromatography to remove the liberated peptides. the peptides remaining associated with the liposomes were then separated from the phospholipid by sephadex lh-60 column chromatography and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. ...19911830591
sequence and structure of mtr, an amino acid transport gene of neurospora crassa.the gene product of the mtr locus of neurospora crassa is required for the transport of neutral aliphatic and aromatic amino acids via the n system. we have previously cloned three cosmids containing neurospora dna that complement the mtr-6(r) mutant allele. the cloned dnas were tightly linked to restriction fragment length polymorphisms that flank the mtr locus. a 2.9-kbp fragment from one cosmid was subcloned and found to complement the mtr-6(r) allele. here we report the sequence of the fragm ...19911838345
rip (repeat induced point mutation) as a tool in the analysis of p-450 and sterol biosynthesis in neurospora crassa. 19911838342
pectinase production by neurospora crassa: purification and biochemical characterization of extracellular polygalacturonase activity.the production of pectinase was studied in neurospora crassa, using the hyperproducer mutant exo-1, which synthesized and secreted five to six times more enzyme than the wild-type. polygalacturonase, pectin lyase and pectate lyase were induced by pectin, and this induction was glucose-repressible. polygalacturonase was induced by galactose four times more efficiently than by pectin; in contrast the activity of lyases was not affected by galactose. the inducing effect of galactose on polygalactur ...19911835496
a novel phenotype of an excision-repair mutant in neurospora crassa: mutagen sensitivity of the mus-18 mutant is specific to uv.a uv-sensitive mutant has been isolated from uv-mutagenized conidia of neurospora crassa. the mutation responsible for the lesion was mapped in linkage group vl, proximal to the nucleolus organizer region. we designated the mutant mus-18. the sensitivity of the mus-18 mutant to uv-irradiation was not particularly high, being less than twice that of the wild-type strain. however, the frequency of mutations at the ad-3 loci induced by uv was extremely high even at low doses, under conditions where ...19911832207
primary structure of the nuclear-encoded 18.3 kda subunit of nadh: ubiquinone reductase (complex i) from neurospora crassa mitochondria.the primary structure of the nuclear-encoded 18.3 kda subunit of the respiratory chain nadh: ubiquinone reductase (complex i) from neurospora crassa was determined by sequencing cdna and the n-terminus of the protein. the cdna contains an open reading frame for a protein of 206 amino acids. the mature protein consists of 173 amino acids and has a molar mass of 18,341 da. the precursor protein includes a characteristic mitochondrial import sequence with a typical matrix peptidase processing site.19911830490
primary structure of the nuclear-encoded 29.9 kda subunit of nadh: ubiquinone reductase from neurospora crassa mitochondria.we isolated and sequenced cdna for the 29.9 kda subunit of mitochondrial nadh: ubiquinone reductase (complex i) from a neurospora crassa library in the lambda gt11 expression vector. the n-terminus of the mature protein was determined by edman-degradation. the cdna contains an open reading frame encoding a preprotein of 273 amino acids. the presequence of the transit protein essential for mitochondrial import is eight residues long. northern-blot analysis shows, that the level of the correspondi ...19911830489
inhibition of neurospora crassa cytosolic chitinase by allosamidin.a cytosolic chitinase (20 kda by sds-page) was partially purified from neurospora crassa. linear kinetics for enzyme activity were obtained using the substrate [3h]-labelled regenerated chitin, the preparation yielding an apparent km of 0.965 mg ml-1 and a vmax of 3.83 micrograms glcnac min-1 (mg protein)-1. the enzyme was highly sensitive to allosamidin, an inhibitor of insect chitinase, exhibiting an ic50 of 1.6 microm. unlike other chitinases that are inhibited by allosamidin, the mode of inh ...19911834520
beta-oxidation system of the filamentous fungus neurospora crassa. structural characterization of the trifunctional protein.treatment of the trifunctional protein from neurospora crassa with various proteases produced almost identical patterns of proteolytic fragments. to study the structural features of the protein in more detail limited proteolysis with trypsin was carried out. polyclonal antibodies were raised against three different tryptic fragments. with the help of immunological methods and amino-terminal sequence analysis we were able to monitor the sequential cleavage steps during proteolysis. two major frag ...19911830049
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
the respiratory response to heat shock in neurospora crassa.a sharp decrease in oxygen uptake occurred in neurospora crassa cells that were transferred from 30 degrees c to 45 degrees c, and the respiration that resumed later at 45 degrees c was cyanide-insensitive. energization of mitochondria, measured in vivo with fluorescence microscopy and a carbocyanine dye, also declined sharply in cells at 45 degrees c. electron microscopy showed no changes in mitochondrial complexity; however, the cytoplasm of heat-shocked cells was deficient in glycogen granule ...19911833266
patch clamping vdac in liposomes containing whole mitochondrial membranes.whole mitochondrial membranes isolated from neurospora crassa were reconstituted into liposomes and patch clamped. clear activity characteristic of the mitochondrial channel vdac was found, namely: open state conductance of 650 ps (in 150 mm kcl, 1 mm cacl2, 20 mm hepes, ph 7.2), voltage-dependent closure at both positive and negative potentials, change in conductance upon channel closure of about 450 ps in response to negative and positive potentials, and increased voltage dependence in the pre ...19911723104
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
gtp is required for the integration of a fragment of the neurospora crassa h(+)-atpase into homologous microsomal vesicles.the integration of a fragment of the neurospora crassa plasma membrane h(+)-atpase was examined to determine if insertion of the fragment into homologous microsomal vesicles is obligatorily dependent on a nucleoside triphosphate. rna transcripts that encoded the amino terminal 344 amino acids of the neurospora crassa plasma membrane h(+)-atpase(pma(344)+) were translated in a n. crassa in vitro system. the pma(344)+ integrated post-translationally into homologous microsomal vesicles independent ...19911711898
the cdna sequence and expression of an ubiquitin-tail gene fusion in neurospora crassa.the genome of neurospora crassa contains at least one natural fusion gene encoding a single ubiquitin (ubi) unit with a 78-amino acid c-terminal extension. the predominantly basic tail sequence corresponds to a highly conserved ribosomal protein identified in other organisms. the 0.7-kb ubi fusion transcript is mainly expressed in germinating conidia and other stages of active cell replication. under starvation conditions attained by nutrient depletion, or after polyamine depletion, the ubi fusi ...19911650731
nit-3, the structural gene of nitrate reductase in neurospora crassa: nucleotide sequence and regulation of mrna synthesis and turnover.the nit-3 gene of the filamentous fungus neurospora crassa encodes the enzyme nitrate reductase, which catalyzes the first reductive step in the highly regulated nitrate assimilatory pathway. the nucleotide sequence of nit-3 was determined and translates to a protein of 982 amino acid residues with a molecular weight of approximately 108 kda. comparison of the deduced nit-3 protein sequence with the nitrate reductase protein sequences of other fungi and higher plants revealed that a significant ...19911829499
effects of heat shock on the level of trehalose and glycogen, and on the induction of thermotolerance in neurospora crassa.neurospora crassa conidiospore germlings exposed to a heat shock (30-45 c) rapidly accumulated trehalose and degraded glycogen, even in the presence of cycloheximide. this phenomenon was also rapidly reversible upon return of the cells at 30 degrees c. trehalose accumulation at 45 degrees c demanded an exogenous source of carbon and either glucose or glycerol fulfilled such requirement. experiments with the cyclic amp-deficient cr-1 mutant suggested that the effects of temperature shifts on treh ...19911645296
isolation of uridine auxotrophs from trichoderma reesei and efficient transformation with the cloned ura3 and ura5 genes.uridine auxotrophs of the filamentous fungus trichoderma reesei have been selected using a positive screening procedure with 5-fluoro orotate. mutants deficient for the orotidine-5'-phosphate decarboxylase gene (ura3 mutants) and for the orotate phosphoribosyl transferase gene (ura5 mutants) have been characterized. the homologous ura3 and ura5 genes have been isolated and used to transform the auxotrophic mutants. transformation efficiency with these homologous systems is very high (greater tha ...19911913875
regulation of laccase synthesis in induced neurospora crassa cultures.rapidly growing cultures of n. crassa do not produce laccase. exposure of this fungus to different inducing agents leads to a de novo biosynthesis of extracellular laccase in vegetative cultures. in this study the induction of laccase after addition of cycloheximide and d-phenylalanine is reported. de novo synthesis of laccase mrna was followed over 96 h after induction. a fast appearance of the message, as well as its presence over a rather long period, indicates a regulation on a transcription ...19911833078
determination of electric parameters of cell membranes by a dielectrophoresis method.marszalek, p., j. j. zielinsky, and m. fikus (1989. bioelectrochem. bioenerg. 22:289-298) have described a novel design for measuring the complete dielectrophoretic spectrum of a single cell. from the analysis of the dielectrophoretic spectrum, the membrane conductivity, sigma membr, and the membrane dielectric permittivity, epsilon membr, of the cell may be determined according to the theory of dielectrophoresis described by sauer, f. a. (1985. interactions between electromagnetic field and cel ...19911831052
photostimulation of conidiation in mutants of neurospora crassa.various mutants of neurospora crassa were screened for light-stimulated conidiation which is a blue light effect and, at least in strain albino-band, is mediated by the flavoprotein nitrate reductase (nr). nr- mutants showed practically no photoconidiation under standard conditions. however, in fusion products of nit-1 (diaphorase activity present, terminal activity missing) plus nit-3 (terminal activity present, diaphorase activity missing), nr activities and photoconidiation were partially res ...19911830899
loss of nad(p)-reducing power and glutathione disulfide excretion at the start of induction of aerial growth in neurospora crassa.when exponentially growing hyphae of neurospora crassa in aerated liquid cultures are filtered and the resulting mycelial mat is exposed to air, aerial hyphae develop and synchronous conidiation is obtained. the hyphae in direct contact with air adhere to each other within minutes and form aerial hyphae during the following 12 h; the hyphae which are not in direct contact with air do not adhere to each other and do not form aerial hyphae. previous data indicated that oxidative stress was generat ...19911827113
an inducible gene expression system for neurospora crassa.neurospora crassa acetyl coa synthetase is highly induced when the growing mycelium is transferred from sucrose- to acetate-based medium. the inducible promoter of this gene has been isolated and used to control the expression of glutamate dehydrogenase. transformants containing this expression cassette show gdh levels up to 25 times higher than the nontransformed host strain. this expression cassette will form the basis of a system of heterologous gene expression.19911369451
regulation of biosynthesis of l-amino acid oxidase by neurospora crassa.l-amino acid oxidase was purified from liquid cultures of neurospora crassa induced by l-phenylalanine, d-phenylalanine, atp and cycloheximide. although the four enzyme species isolated were found to differ in size and electrophoretic mobility, they were the product of a single gene as demonstrated by genomic southern analysis. northern analysis of total cellular rna showed a rapid increase of l-amino acid oxidase mrna in response to the different inducing agents studied. these data suggest that ...19911829425
metalloselenonein, the selenium analogue of metallothionein: synthesis and characterization of its complex with copper ions.we used an automated peptide synthesizer to produce a peptide, metalloselenonein, that contains selenocysteine residues substituted for all cysteine residues in neurospora crassa copper metallothionein. metalloselenonein binds 3 mol of cu(i) per mol. this adduct shows a broad absorption band between 230 and 400 nm and a fluorescence band at 395 nm, which can be attributed to copper-selenolate coordination. the circular dichroism spectrum of the copper-metalloselenonein complex shows a positive b ...19911826562
ca2+ calmodulin-dependent protein kinase activity in the ascomycetes neurospora crassa.deae-cellulose column chromatography of neurospora crassa soluble mycelial extracts leads to the resolution of three major protein kinase activity peaks designated pki, pkii, and pkiii. pkii activity is stimulated by ca2+ and neurospora or brain calmodulin. maximal stimulation was observed at 2 microm-free ca2+ and 1 microgram/ml of the modulator. the stimulatory effect of the ca(2+)-calmodulin complex was blocked by egta and by some calmodulin antagonists such as phenothiazine drugs or compound ...19911652680
regulation of molybdenum cofactor species in the green alga chlamydomonas reinhardtii.molybdenum cofactor (moco) of molybdoenzymes is constitutively produced in cells of the green alga chlamydomonas reinhardtii grown in ammonium media, under which conditions certain molybdoenzymes are not synthesized. in soluble form, moco was found to be present in several forms: (i) as a low mr free species; (ii) bound to a moco-carrier protein of about 50 kda that could release moco to directly reconstitute in vitro nitrate reductase activity in the nit-1 mutant of neurospora crassa, but not t ...19911826614
nucleotide and dna uptake by neurospora crassa: involvement of an uptake stimulating protein.the basal and dusf (dna-uptake-stimulating factor, described previously by schablik and szabó (1981) fems microbiol. lett. 10, 395-397) stimulated uptake of [3h]dna and radioactive nucleotides by neurospora crassa (fgsc 1118, slime) cell-wall-less strain was studied. the uptake of [3h]dna by the cells is a saturable and time-dependent process. the ph and temperature optimum for [3h]dna uptake are ph 7 and 27 degrees c, respectively. both basal and dusf-stimulated uptake of [3h]dna are inhibited ...19911826455
specificity of repeat-induced point mutation (rip) in neurospora: sensitivity of non-neurospora sequences, a natural diverged tandem duplication, and unique dna adjacent to a duplicated region.the process designated rip (repeat-induced point mutation) alters duplicated dna sequences in the sexual cycle of neurospora crassa. we tested whether non-neurospora sequences are susceptible to rip, explored the basis for the observed immunity to this process of a diverged tandem duplication that probably arose by a natural duplication followed by rip (the neurospora zeta-eta region), and investigated whether rip extends at all into unique sequences bordering a duplicated region. bacterial sequ ...19911827630
recurrence of repeat-induced point mutation (rip) in neurospora crassa.duplicate dna sequences in the genome of neurospora crassa can be detected and mutated in the sexual phase of the life cycle by a process termed rip (repeat-induced point mutation). rip occurs in the haploid nuclei of fertilized, premeiotic cells before fusion of the parental nuclei. both copies of duplications of gene-sized sequences are affected in the first generation at frequencies of approximately 50-100%. we investigated the extent to which sequences altered by rip remain susceptible to th ...19911827629
surface topography and molecular stoichiometry of the mitochondrial channel, vdac, in crystalline arrays.the mitochondrial outer membrane contains a protein, called vdac, that forms large aqueous pores. in neurospora crassa outer membranes, vdac forms two-dimensional crystalline arrays whose size and frequency can be greatly augmented by lipase treatment of these membranes (c. mannella, science 224, 165, 1984). fourier filtration and surface reconstruction of freeze-dried/shadowed (45 degrees) arrays produced detailed images of two populations of crystals, whose lattices are mirror images of each o ...19911725124
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