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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
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
characterization of neurospora cpc1, a bzip dna-binding protein that does not require aligned heptad leucines for dimerization.cpc1 is the transcriptional activator of amino acid biosynthetic genes of neurospora crassa. cpc1 function in vivo was abolished upon deletion of segments of cpc-1 corresponding to the presumed transcription activation domain, the dna-binding and dimerization domains, or a 52-residue connector segment of cpc1. a truncated cpc1 polypeptide containing only the carboxy-terminal 57-residue segment of cpc1 was sufficient to form homodimers that bound dna. however, deletion of the segment of cpc-1 cor ...19911824960
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
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
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
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
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
isolation and characterization of the adenylate cyclase structural gene of neurospora crassa.a single gene (nac) encoding an adenylate cyclase was cloned from the genomic dna library of neurospora crassa, using the dna fragment encoding the catalytic domain of adenylate cyclase of saccharomyces cerevisiae as a probe. the open reading frame of this gene (6900 base pairs) was interrupted three time by introns. the protein encoded consists of 2300 amino acids and has adenylate cyclase activity. n. crassa adenylate cyclase has a high degree of homology with the catalytic domains of yeast an ...19911680356
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
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
expression of the escherichia coli beta-glucuronidase gene in pseudocercosporella herpotrichoides.the plant-pathogenic fungus pseudocercosporella herpotrichoides has been successfully transformed by using two different positive selection systems in combination with the escherichia coli gusa gene. the selectable markers used in this study were the hygromycin b phosphotransferase gene (hph) from e. coli and the gene (bml) for beta-tubulin from a benomyl-resistant mutant of neurospora crassa. a lower transformation rate was obtained with the bml system than with the hph system. conversely, cotr ...19911746951
mutagenic potency and specificity of procarbazine in the ad-3 forward-mutation test in growing cultures of heterokaryon 12 of neurospora crassa.procarbazine (natulan) was tested for its mutagenic potency and specificity in the ad-3 forward-mutation test in heterokaryon 12 (h-12) of neurospora crassa. in these experiments, procarbazine was a weak mutagen when present in growing cultures but nonmutagenic when conidial suspensions (nongrowing conidia) were treated. a total of 208 ad-3 mutants recovered after exposure of growing cultures of h-12 to 1 mg of procarbazine/ml, and 2 ad-3 mutants of spontaneous origin, were characterized genetic ...19911824718
catalysis of protein folding by cyclophilins from different species.cyclophilins are a class of ubiquitous proteins with yet unknown function. they were originally discovered as the major binding proteins for the immunosuppressant cyclosporin a. the only known catalytic function of these proteins in vitro is the cis/trans isomerization of xaa-pro bonds in oligopeptides. this became clear after the discovery that bovine cyclophilin is identical with porcine prolyl isomerase. this enzyme accelerates slow, proline-limited steps in the refolding of several proteins. ...19911825312
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
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
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
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
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
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
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
[synthesis and antimicrobial activity of substituted fluorobenzyl benzylidenethiazolidinediones and imidazolidinediones].the synthesis of six benzylidene thiazolidine-diones and three benzylidene imidazolidine-diones is described. in order to investigate their antimicrobial activity, they are evaluated against micro-organism such as staphylococcus aureus, streptococcus feacalis, mycobacterium smegmatis and neurospora crassa.19911795213
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
relationship of the membrane atpase from halobacterium saccharovorum to vacuolar atpases.polyclonal antiserum against subunit a (67 kda) of the vacuolar atpase from neurospora crassa reacted with subunit i (87 kda) from a membrane atpase of the extremely halophilic archaebacterium halobacterium saccharovorum. the halobacterial atpase was inhibited by nitrate and n-ethylmaleimide; the extent of the latter inhibition was diminished in the presence of adenosine di- or triphosphates. 4-chloro-7-nitrobenzofurazan inhibited the halobacterial atpase also in a nucleotide-protectable manner; ...19911824911
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
isolation and sequence analysis of a beta-tubulin gene from aspergillus flavus and its use as a selectable marker.an altered beta-tubulin gene that confers resistance to benomyl [whose active ingredient is 2-(methoxycarbonylamino)benzimidazole (mbc)] was isolated from a dna library of aspergillus flavus and used as a selectable marker for transformation. the beta-tubulin gene was cloned into a plasmid vector containing the pyr-4 gene of neurospora crassa, and transformants were selected either for uracil prototrophy or mbc resistance. transformants selected for uracil prototrophy were of three phenotypic cl ...19902128007
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
paraquat toxicity and pyridine nucleotide coenzyme synthesis: a data correction.the decrease in pyridine nucleotide coenzymes which occurs during poisoning of escherichia coli by hyperbaric oxygen or paraquat is not due to impairment of nicotinatemononucleotide pyrophosphorylase (carboxylating) [ec 2.4.2.19] as was previously proposed (brown, o.r. et al. biochem. biophys. res. commun. 91:982-990; 1979). this was shown directly using extracts of e. coli, prepared after exposure to 1 mm paraquat or 4.2 atmospheres of oxygen. the enzyme also was not impaired in neurospora cras ...19902139629
two complex regions, including a tata sequence, are required for transcription by rna polymerase i in neurospora crassa.in order to define the rna polymerase i transcriptional apparatus and how it might interact with regulatory signals, the dna sequences necessary for 40s rrna transcription in neurospora crassa were determined. a systematic set of deletion, substitution and insertion mutations were assayed in a homologous in vitro system. the sequences required for transcription of the gene consist of two large domains (i and ii) from -113 to -37, and -29 to +4, respectively. complete deletion of either domain ab ...19902139932
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
molybdenum cofactor requirement for in vitro activation of apo-molybdoenzymes of escherichia coli.the apo-nitrate reductase precursor in an escherichia coli chlb mutant preparation obtained following growth in the presence of tungstate is activated by incubation with protein fa and a heat-treated preparation from an e. coli crude extract. we show that the requirement for heat-treated e. coli crude extract can be fulfilled by material obtained from either of two heat-denatured purified e. coli molybdoenzymes, namely nitrate reductase or trimethylamine n-oxide reductase. apo-trimethylamine n-o ...19902141097
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
[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
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
molecular cloning of subunits of complex i from neurospora crassa. primary structure and in vitro expression of a 22-kda polypeptide.a lambda gt11 cdna expression library was screened with antibodies directed against individual subunits of complex i from neurospora crassa. clones encoding cytoplasmically synthesized polypeptides with apparent molecular masses of 22, 29, 31, and 33 kda were isolated. northern blot analysis revealed that the corresponding genes are transcribed into mrna species of about 0.85, 0.95, 1.3, and 1.4 kilobases, respectively. further characterization of clones encoding the 22-kda subunit was performed ...19902142943
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
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
evidence for the presence of a proton pump of the vacuolar h(+)-atpase type in the ruffled borders of osteoclasts.microsomal membrane vesicles prepared either from chicken medullary bone or isolated osteoclasts were shown to have atp-dependent h(+)-transport activity. this activity was n-ethylmaleimide-sensitive but resistant to oligomycin and orthovanadate, suggesting a vacuolar-type atpase. furthermore, immunological cross-reactivity of 60- and 70-kd osteoclast membrane antigens with neurospora crassa vacuolar atpase was observed when analyzed by immunoblotting. same antibodies labeled only osteoclasts in ...19902144003
nit-2, the major positive-acting nitrogen regulatory gene of neurospora crassa, encodes a sequence-specific dna-binding protein.the nit-2 major nitrogen regulatory gene of neurospora crassa turns on the expression of various unlinked structural genes that specify nitrogen-catabolic enzymes under nitrogen-limitation conditions. the nit-2 gene encodes a protein of 1036 amino acid residues with a single zinc finger and a downstream basic region that may make up a dna-binding domain. the zinc-finger domain of the nit2 protein was synthesized in two ways to examine its dna-binding activity with gel-band-mobility shift and dna ...19902142530
carotenoid desaturases from rhodobacter capsulatus and neurospora crassa are structurally and functionally conserved and contain domains homologous to flavoprotein disulfide oxidoreductases.the characteristic red color of some photosynthetic bacteria and the orange color of neurospora conidia is due to the presence of carotenoids, photoprotective pigments synthesized by plants, algae, bacteria, and fungi. generally, carotenoids are tetraterpenes in which absorption of visible light and photoprotection are mediated by a chain of conjugated double bonds, the chromophore, which is formed by successive desaturations of phytoene, a colorless precursor. the genes al-1 and crti mediate th ...19902144293
the neurospora crassa arg-2 locus. structure and expression of the gene encoding the small subunit of arginine-specific carbamoyl phosphate synthetase.we have characterized genomic and cdna clones for arg-2, the gene encoding the small subunit of the neurospora crassa arginine-specific carbamoyl phosphate synthetase (cps-a), and examined its transcriptional regulation. the polypeptide's predicted amino acid sequence (453 residues) is 56% and 36% identical with the sequences of the homologous polypeptides of saccharomyces cerevisiae and escherichia coli, respectively. the arg2 polypeptide has an additional amino-terminal domain with the hallmar ...19902141606
premeiotic disruption of the neurospora crassa malate synthase gene by native and divergent dnas.repeat-induced point mutation (rip) has been used to generate new mutations in the previously uncharacterised gene for malate synthase in neurospora crassa. molecular clones carrying the am (nadp-glutamate dehydrogenase) gene and the malate synthase gene from either n. crassa or aspergillus nidulans have been introduced into neurospora as ectopic duplicate copies by transformation, selecting for the am+ function in a deletion host. a number of meiotic progeny derived from these transformants wer ...19901979142
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
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
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
use of nuclear dna restriction fragment length polymorphisms to analyze the diversity of the aspergillus flavus group: a. flavus, a. parasiticus, and a. nomius.recombinant dna clones carrying high-copy or low-copy sequences from aspergillus nidulans and neurospora crassa were used to identify restriction fragment length polymorphisms (rflps) diagnostic for members of the a. flavus group: a. flavus, a. parasiticus, and a. nomius. these fungi were resolved into three distinct categories when they were grouped according to rflp patterns. subgroups within these categories were also evident. this limited rflp analysis of nuclear dna of members of the a. fla ...19901976300
x-ray-induced specific-locus mutations in the ad-3 region of two-component heterokaryons of neurospora crassa. v. irreparable mutants of genotype ad-3a ad-3b, ad-3a ad-3b nic-2, and ad-3b nic-2 result from multilocus deletion and an unexpectedly high frequency of multiple-locus mutations.more extensive complementation tests than those performed initially (webber and de serres, 1965) on a series of 832 x-ray-induced specific-locus mutations in the adenine-3 (ad-3) region of a two-component heterokaryon (h-12) of neurospora crassa (de serres, 1989a) showed that unexpectedly high frequencies of specific-locus mutations in the ad-3 region have additional, but separate, sites of recessive lethal (rlcl) damage in the immediately adjacent genetic regions. the frequencies of these x-ray ...19902138248
deletion analysis of domain independence in the trp1 gene product of neurospora crassa.the trifunctional trp1 gene from neurospora crassa (n-trp1) was subcloned into the yeast-escherichia coli shuttle vector yep13 and expressed in saccharomyces cerevisiae. the three activities of the n-trp1 gene product were detected in yeast mutants that lacked either n-(5'-phosphoribosyl) anthranilate (pra) isomerase or both the glutamine amidotransferase function of anthranilate synthase and indole-3-glycerol phosphate (ingp) synthase. the protein was detected on immunoblots only as the full le ...19902147978
a neurospora crassa ribosomal protein gene, homologous to yeast cry1, contains sequences potentially coordinating its transcription with rrna genes.we have isolated and sequenced a neurospora crassa ribosomal protein gene (designated crp-2) strongly homologous to the rp59 gene (cry1) of yeast and the s14 ribosomal protein gene of mammals. the inferred sequence of the crp-2 protein is more homologous (83%) to the mammalian s14 sequence than to the yeast rp59 sequence (69%). the gene has three intervening sequences (ivss) two of which are offset 7 bp from the position of ivss in the mammalian genes. none correspond to the position of the ivs ...19901977135
tests for the genotoxicity of m-amsa, etoposide, teniposide and ellipticine in neurospora crassa.the antitumor agents m-amsa, etoposide, teniposide and ellipticine have been reported to be potent clastogens in mammalian cells but non- or weakly mutagenic in bacteria; these observations have been correlated to the interference of these chemicals with dna topoisomerase ii activity in the former, but not in the latter, organisms. the genotoxicity of these 4 agents was evaluated using ad-3 reverse- and forward-mutation tests in neurospora crassa. these agents (up to 0.8 mumole/plate) did not ca ...19902137196
mating type and mating strategies in neurospora.in the heterothallic species neurospora crassa, strains of opposite mating type, a and a, must interact to give the series of events resulting in fruiting body formation, meiosis, and the generation of dormant ascospores. the mating type of a strain is specified by the dna sequence it carries in the mating type region; strains that are otherwise isogenic can mate and produce ascospores. the dna of the a and a regions have completely dissimilar sequences. probing dna from strains of each mating t ...19902140508
deoxyhypusine/hypusine formation on a 21,000-dalton cellular protein in a neurospora crassa mutant in vivo and in vitro.hypusine formation on an 18,000-dalton cellular protein is a unique spermidine-dependent, post-translational modification that appears to be ubiquitous in mammalian cells. to determine whether this modification also exists in lower eukaryotes, we examined possible labeling in vitro and in vivo of cellular protein(s) by [3h]spermidine in a mutant strain of neurospora crassa (arge-12 ota aga) in which ornithine and polyamine synthesis could be nutritionally manipulated. because of poor uptake of p ...19902137713
site-directed mutagenesis of the 'zinc finger' dna-binding domain of the nitrogen-regulatory protein nit2 of neurospora.the major nitrogen-regulatory gene nit-2 of neurospora crassa activates the expression of numerous unlinked structural genes which specify nitrogen-catabolic enzymes during conditions of nitrogen limitation. the nit-2 gene encodes a regulatory protein of 1036 amino acid residues with a single 'zinc finger' and a downstream basic region, which together may constitute a dna-binding domain. the zinc finger domain of the nit2 protein was synthesized in vitro and also expressed as a fusion protein in ...19902150539
remnants of an ancient pathway to l-phenylalanine and l-tyrosine in enteric bacteria: evolutionary implications and biotechnological impact.the pathway construction for biosynthesis of aromatic amino acids in escherichia coli is atypical of the phylogenetic subdivision of gram-negative bacteria to which it belongs (r. a. jensen, mol. biol. evol. 2:92-108, 1985). related organisms possess second pathways to phenylalanine and tyrosine which depend upon the expression of a monofunctional chorismate mutase (cm-f) and cyclohexadienyl dehydratase (cdt). some enteric bacteria, unlike e. coli, possess either cm-f or cdt. these essentially c ...19902082822
primary structure of the non-transcribed spacer region and flanking sequences of the ribosomal dna of neurospora crassa and comparison with other organisms.the non-transcribed spacer (nts) region of the rdna of neurospora crassa contains the transcription regulatory sequences. we isolated a 3.4 kb ecori fragment from wild type n.crassa rdna and cloned in the plasmid pbr325 at the ecori site. the insert contains the entire nts region along with the flanking sequences. nucleotide sequencing of 3592 nt shows many interesting features like: the nts region is rich in g+c content (65% g+c); it contains the conserved rrna processing site 6 (with the nucle ...19902142594
neurospora endo-exonuclease is immunochemically related to the recc gene product of escherichia coli.immunochemical cross-reaction between the endo-exonuclease of neurospora crassa, an enzyme previously implicated in recombination and recombinational dna repair, and the recc-encoded polypeptide of escherichia coli has been detected by immunoblotting extracts of strains of e. coli having a deletion that includes the recbcd genes but carrying multicopy plasmids bearing all three of the recbcd genes or only one or two of these genes. it was predicted that homology would also be found at the amino ...19902152915
cys-3, the positive-acting sulfur regulatory gene of neurospora crassa, encodes a sequence-specific dna-binding protein.cys-3, the positive-acting master sulfur regulatory gene of neurospora crassa, turns on the expression of an entire set of unlinked structural genes which encode sulfur-catabolic enzymes. cys-3 encodes a protein of 236 amino acid residues and contains a potential bipartite dna-binding domain which consists of a leucine zipper and an adjacent highly basic region. gel band mobility shift and dna footprint experiments were used to demonstrate that the cys3 protein, expressed in escherichia coli, bi ...19902142156
aberrant mitochondrial processing of chimaeric import precursors containing subunits 8 and 9 of yeast mitochondrial atp synthase.a set of chimaeric precursors which contain the same leader sequences but different passenger proteins has been analyzed for the site of protease cleavage following import into yeast mitochondria. each precursor comprises the leader of neurospora crassa subunit 9 of mitochondrial atp synthase fused to subunit 8 or 9 of the corresponding yeast enzyme. precursors containing the first five residues of mature n. crassa subunit 9 interposed between the leader and the yeast passenger protein were clea ...19902151020
regulation of inorganic sulfate activation in filamentous fungi. allosteric inhibition of atp sulfurylase by 3'-phosphoadenosine-5'-phosphosulfate.atp sulfurylases from penicillium chrysogenum, penicillium duponti, aspergillus nidulans, and neurospora crassa are strongly inhibited by 3'-phosphoadenosine-5'-phosphosulfate (paps), the product of the second (adenosine-5'-phosphosulfate kinase-catalyzed) reaction in the two-step activation of inorganic sulfate. the v versus [paps] plots are sigmoidal. at physiological concentrations of mgatp (0.17-3 mm) and so4(2-) (0.4-10 mm), the [i]0.5 for paps inhibition of the p. chrysogenum enzyme is 35- ...19902162344
nucleotide sequence and analysis of nmr, a negative-acting regulatory gene in the nitrogen circuit of neurospora crassa.in neurospora the expression of a set of unlinked structural genes, which allows utilization of various nitrogen-containing compounds, is controlled by the positive-acting nit-2 gene and the negative-acting nmr gene. the nucleotide sequence of the nmr gene has been determined and a long open reading frame which encodes a putative protein of 54854 daltons has been identified. a full-length cdna clone was obtained and its the sequence revealed that the nmr gene contains no introns. the transcripti ...19902146484
import into mitochondria of precursors containing hydrophobic passenger proteins: pretreatment of precursors with urea inhibits import.we have studied the import into isolated yeast mitochondria of three hydrophobic passenger proteins attached to the n-terminal cleavable presequence of mitochondrial atpase subunit 9 from neurospora crassa. one natural precursor (pn9) contained n. crassa subunit 9; two chimaeric precursors, n9l/y8-1 and n9l/y9-2, respectively contained yeast mitochondrial atpase subunits 8 and 9. in the absence of urea, pn9 and n9l/y8-1 are imported efficiently but n9l/y9-2 is not imported. after pretreatment of ...19902168755
molecular analysis of nuc-1+, a gene controlling phosphorus acquisition in neurospora crassa.in response to phosphorus starvation, neurospora crassa makes several enzymes that are undetectable or barely detectable in phosphate-sufficient cultures. the nuc-1+ gene, whose product regulates the synthesis of these enzymes, was cloned and sequenced. the nuc-1+ gene encodes a protein of 824 amino acids with a predicted molecular weight of 87,429. the amino acid sequence shows homology with two yeast proteins whose functions are analogous to that of the nuc-1 protein. two nuc-1+ transcripts of ...19902146493
spatial and biological characterisation of the complete quinic acid utilisation gene cluster in aspergillus nidulans.heterologous probing of restriction digests of chromosomal dna from aspergillus nidulans with radioactively labelled probes encoding dehydroshikimate dehydratase (qa-4) and a repressor gene (qa1-s) from neurospora crassa revealed a pattern of hybridisation inconsistent with an equivalent single copy of each gene in a. nidulans. screening of size-selected and total genome a. nidulans dna libraries allowed the isolation of four unique classes of sequence, two of which hybridised to the qa-4 probe, ...19902175387
molecular structure of a gene, vma1, encoding the catalytic subunit of h(+)-translocating adenosine triphosphatase from vacuolar membranes of saccharomyces cerevisiae.subunit a of the vacuolar membrane h(+)-translocating adenosine triphosphatase of the yeast saccharomyces cerevisiae contains a catalytic site for atp hydrolysis. n-terminal sequences of six tryptic peptides of the subunit were determined. based on the peptide sequence information, a 39-base oligonucleotide probe was synthesized, and the gene encoding the subunit (vma1) was isolated from a genomic dna library by hybridization. the nucleotide sequence of the gene predicts a polypeptide of 1,071 a ...19902139027
the general mitochondrial matrix processing protease from rat liver: structural characterization of the catalytic subunit.a critical step in the import of nuclear-encoded precursor proteins into mitochondria involves proteolytic cleavage of their amino-terminal leader peptides by processing proteases found in the mitochondrial matrix. we report here the characterization of the general matrix processing protease from rat liver mitochondria. the final enzyme preparation consisted of two polypeptides, a catalytically active 55-kda subunit and a 52-kda one. to deduce the complete primary structure of the 55-kda subunit ...19902236012
[genetic engineering in filamentous fungi: cloning of the invertase gene from neurospora crassa].the invertase wild type gene of n. crassa was cloned into the yrp7 yeast vector. this recombinant plasmid was selected by functional complementation of an invertaseless mutant strain of s. cerevisiae. the isolated recombinant plasmid (named pnc2) carries a 7.6 kb bamhi dna fragment from n. crassa. the cloned dna hybridized with the n. crassa genomic dna and transformed an invertase mutant of n. crassa inv- to inv+. transformation of n. crassa inv- to inv+ seems to take at least two different int ...19902151941
mycenon, a new metabolite from a mycena species ta 87202 (basidiomycetes) as an inhibitor of isocitrate lyase.mycenon (c11h5cl3o3), a new inhibitor of isocitrate lyase (ec 4.1.3.1) was isolated from the culture broth of a basidiomycete, mycena sp. mycenon is a novel chlorinated benzoquinone derivative which is also active against bacteria and fungi. malate synthase (ec 4.1.3.2) the second key enzyme of the glyoxylate cycle was not affected by mycenon. isocitrate lyase preparations from plants, bacteria and fungi were sensitive. the following ki-values for mycenon have been determined: ricinus communis, ...19902258323
expression in saccharomyces cerevisiae of a gene associated with cytoplasmic male sterility from maize: respiratory dysfunction and uncoupling of yeast mitochondria.we asked whether the mitochondrial t-urf13 gene, associated with the male sterility phenotype of t cytoplasm in maize, can be expressed in saccharomyces cerevisiae and whether this expression can mimic the effects observed in maize. we introduced the universal code equivalent of the t-urf13 gene into the s. cerevisiae nucleus by transformation and directed its translation product into mitochondria by means of a fusion with the targeting presequence from neurospora crassa at-pase subunit 9. we sh ...19902259341
an evolutionary comparison of acinetobacter calcoaceticus trpf with trpf genes of several organisms.the deduced amino acid sequence of acinetobacter calcoaceticus n-(5'-phosphoribosyl) anthranilate isomerase (prai), which is coded by trpf, was compared with trpf of caulobacter crescentus, escherichia coli, bacillus subtilis, saccharomyces cerevisiae, neurospora crassa, and aspergillus nidulans. sixty percent of identical or similar amino acids were located in alpha/beta tim (triose-phosphate isomerase) barrels and in residues important in substrate binding and catalysis. in addition, the analy ...19902299982
cloning of a saccharomyces cerevisiae gene encoding a protein homologous to allantoicase of neurospora crassa. 19911839481
the 49 k subunit of nadh: ubiquinone reductase (complex i) from neurospora crassa mitochondria: primary structure of the gene and the protein.the primary structure of the 49 k subunit of the respiratory chain nadh:ubiquinone reductase (complex i) from neurospora crassa was determined by sequencing cdna, genomic dna and the n-terminus of the mature protein. the sequence lengths correlate to a molecular mass of 54,002 daltons for the preprotein and 49,239 daltons for the mature protein. the presequence consists of 42 amino acids of typical composition for sequences which target nuclear-encoded proteins into mitochondria. the mature prot ...19902147127
endo-exonuclease of aspergillus nidulans.endo-exonuclease (ee) has been found in both active and inactive, but trypsin-activatable, forms in aspergillus nidulans. active ee was present mainly in nuclei, mitochondria, and vacuoles, while trypsin-activatable ee was mainly in the cytosol. the active form accounts for over 90% of the neutral deoxyribonuclease activity extracted from mycelia. a single strand (ss) dna-binding ee associated with a 28 kilodalton (kda) polypeptide was partially purified and characterized. it was found to closel ...19902161674
neurospora crassa a mating-type region.the mating-type locus of the haploid filamentous fungus neurospora crassa is a regulatory region that controls entry into the sexual cycle and prevents formation of mixed mating-type heterokaryons in the vegetative phase. the locus consists of alternative sequences called a and a. the a mating-type dna sequence of neurospora crassa is composed of a region of 5301 base pairs that has little similarity to the sequence present at the mating-type locus in an a mating-type strain. however, the sequen ...19902142303
neurospora crassa a mating-type region.the a mating-type region of neurospora crassa controls several major events in both the sexual and asexual phases of the fungal life cycle. this 3235-base-pair dna segment is not homologous to the comparable genetic region of the a mating type. the unique a and a regions are bordered by nearly identical dna sequences. the a genetic region contains at least two functional segments. one segment encodes a perithecium maturation function that is dependent on the second segment for phenotypic express ...19902142304
mitochondrial dna sequence analysis of the cytochrome oxidase subunit ii gene from podospora anserina. a group ia intron with a putative alternative splice site.a 5 kb region of the 95 kb mitochondrial genome of podospora anserina race s has been mapped and sequenced (1 kb = 10(3) base-pairs). this dna region is continuous with the sequence for the nd4l and nd5 gene complex in the accompanying paper. we show that this sequence contains the gene for cytochrome oxidase subunit ii (coii). this gene is 4 kb in length and is interrupted by a subgroup ib intron (1267 base-pairs (bp) in length) and a subgroup ia intron (1992 bp in length). this group ia intron ...19902157023
simultaneous purification of three mitochondrial enzymes. acetylglutamate kinase, acetylglutamyl-phosphate reductase and carbamoyl-phosphate synthetase from neurospora crassa.the early enzymes of arginine biosynthesis in neurospora crassa are localized in the mitochondrion and catalyze the conversion of glutamate to citrulline. the final conversion of citrulline to arginine occurs via two enzymatic steps in the cytoplasm. we have devised a method for the isolation and purification of three of the mitochondrial arginine biosynthetic enzymes from a single extract. acetylglutamate kinase and acetylglutamyl-phosphate reductase (both products of the complex arg-6 locus) w ...19862420793
h+/atp stoichiometry of proton pumps from neurospora crassa and escherichia coli.a kinetic method has been used to measure the apparent stoichiometry of h+ ions translocated per atp split by membrane-bound [h+]-atpases. in this method, membrane vesicles are suspended in well-buffered medium, atp is added, and a fluorescent probe of delta ph (acridine orange) is used to detect the formation of a steady-state ph gradient. at the steady state, it is assumed that proton pumping in one direction is exactly balanced by the leak of protons in the opposite direction. the pump is the ...19862425739
heterozygous effects of x-ray-induced specific locus mutations in the ad-3 region of neurospora crassa: implications for human genetic risk assessment. 19902143295
gamma-ray-sensitive mutants of neurospora crassa with characteristics analogous to ataxia telangiectasia cell lines.well characterized gamma-ray sensitive mutants of the fungus neurospora crassa have been screened for characteristics analogous to those of cell lines derived from humans with the genetic disease, ataxia telangiectasia (at). two neurospora mutants, uvs-6 and mus-9, show the at cell line characteristics of gamma-ray and bleomycin sensitivity, and little or no repression of dna synthesis following treatment with these agents. normal human or neurospora cells show an extensive biphasic dna synthesi ...19872434849
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
purification of chorismate synthase from a cell culture of the higher plant corydalis sempervirens pers.chorismate synthase (ec 4.6.1.4) was purified from a cell suspension culture of corydalis sempervirens almost 1000-fold to near homogeneity. the subunit mr estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate was 41,900. the mr of the native enzyme was estimated to be 80,100 by gel filtration, suggesting a dimeric structure. antisera directed against the 41.9-kda protein also reacted with the native enzyme. further confirmation of the identity of the purified ...19902146922
purification and further characterization of the second nitrate reductase of escherichia coli k12.two nitrate reductases, nitrate reductase a and nitrate reductase z, exist in escherichia coli. the nitrate reductase z enzyme has been purified from the membrane fraction of a strain which is deleted for the operon encoding the nitrate reductase a enzyme and which harbours a multicopy plasmid carrying the nitrate reductase z structural genes; it was purified 219 times with a yield of about 11%. it is an mr-230,000 complex containing 13 atoms iron and 12 atoms labile sulfur/molecule. the presenc ...19902139607
pneumocystis carinii: sequence from ribosomal rna implies a close relationship with fungi.pneumocystis carinii is the etiologic agent of a lethal pneumonia which occurs in patients with the acquired immune deficiency syndrome (aids) and other immunocompromised hosts. the basic biochemical and genetic characteristics of p. carinii are poorly understood and its taxonomic classification as a protozoan is uncertain. to address the taxonomic question, a method was developed for the extraction of total rna from p. carinii. denaturing agarose gel electrophoresis showed the two ribosomal rna ...19892470612
comparison and cross-species expression of the acetyl-coa synthetase genes of the ascomycete fungi, aspergillus nidulans and neurospora crassa.the genes encoding the acetate-inducible enzyme acetyl-coenzyme a synthetase from neurospora crassa and aspergillus nidulans (acu-5 and faca, respectively) have been cloned and their sequences compared. the predicted amino acid sequence of the aspergillus enzyme has 670 amino acid residues and that of the neurospora enzyme either 626 or 606 residues, depending upon which of the two possible initiation codons is used. the amino acid sequences following the second alternative aug show 86% homology ...19901972535
the beta-tubulin gene of epichloë typhina from perennial ryegrass (lolium perenne).epichloë typhina is a biotrophic fungal pathogen which causes choke disease of pooid grasses. the anamorphic state, acremonium typhinum, is placed in the section albo-lanosa along with related, mutualistic, seed-disseminated endophytes. as an initial study of gene structure and evolution in epichloë and related endophytes, the beta-tubulin gene, tub2, of the perennial ryegrass choke pathogen (etprg) was cloned and sequenced. the coding sequence and the predicted beta-tubulin amino acid sequence ...19902147581
neurospora crassa cdna clones coding for a new member of the ras protein family.a new member of the ras gene family was characterized from neurospora crassa cdna libraries. the clone designated nc-ras codes for a polypeptide containing 213 amino acids (mr 24,000). this polypeptide is 84% homologous to the h-ras-1 domain comprising the first 80 amino acids and 60% homologous to the next 84 residues. the nc-ras polypeptide contains all the well-known sequences involved in the interaction with gtp/gdp, the recognition of the y13-259 neutralizing antibody, the 'effector site' f ...19902146163
genetic transformation system for the aflatoxin-producing fungus aspergillus flavus.a heterologous transformation system was developed for aspergillus flavus with efficiencies greater than 20 stable transformants per micrograms of dna. protoplasts of uracil-requiring strains of the fungus were transformed with plasmid and cosmid vectors containing the pyr-4 gene of neurospora crassa. transformants were selected for their ability to grow and sporulate on medium lacking uracil. vector dna appeared to integrate randomly into the genome of a. flavus with a tendency for multiple, ta ...19892495764
atp-induced protyrosinase synthesis and carboxyl-terminal processing in neurospora crassa.the effects of 3'-5' cyclic amp and atp upon tyrosinase induction in neurospora crassa were examined. northern analysis of total cellular rna revealed rapid de novo synthesis of protyrosinase after addition of these substances to stationary-phase mycelia. the maturation of protyrosinase in crude extracts of mycelia was followed by western analysis. polyclonal rabbit antiserum directed against the denatured carboxyl-terminal extension of protyrosinase does recognize the proform and several interm ...19902150229
structure, exon pattern, and chromosome mapping of the gene for cytosolic copper-zinc superoxide dismutase (sod-1) from neurospora crassa.a 4.8-kilobase bamhi-hindiii fragment encoding the entire neurospora crassa cuzn superoxide dismutase gene (herein designated sod-1) was isolated from a genomic library using two 60-base deoxyoligonucleotide probes corresponding to the published n. crassa amino acid sequence. the nucleotide sequence of the gene encodes an amino acid sequence matching the published protein sequence at 152 of 153 positions. codon preference shows an unusually strong bias such that only 32 of the possible 61 codons ...19902146266
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