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isolation and characterization of a calmodulin-encoding cdna from the pathogenic fungus histoplasma capsulatum.we describe in this paper the isolation and complete sequence of a calmodulin (cam) encoding cdna from the dimorphic pathogenic fungus histoplasma capsulatum (genbank accession u12505). the deduced amino acid sequence was identical to the cam of aspergillus nidulans and had only one amino acid difference from the cam of neurospora crassa. saccharomyces cerevisiae cam, however, has only about 60% amino acid identity compared with h. capsulatum. these data further support the close relationship of ...19968803795
secondary and tertiary structure changes of reconstituted p-glycoprotein. a fourier transform attenuated total reflection infrared spectroscopy analysis.the structure of purified p-glycoprotein functionally reconstituted into liposomes was investigated by attenuated total reflection fourier transform infrared spectroscopy. a quantitative evaluation of the secondary structure and a kinetic of 2h/h exchange of the p-glycoprotein were performed both in the presence and in the absence of mgatp, mgatp-verapamil, and mgadp. this approach was previously shown to be a useful tool to detect tertiary structure changes resulting from the interaction betwee ...19968798727
functional dissection and site-directed mutagenesis of the structural gene for nad(p)h-nitrite reductase in neurospora crassa.neurospora crassa nad(p)h-nitrite reductase, encoded by the nit-6 gene, is a soluble, alpha2-type homodimeric protein composed of 127-kda polypeptide subunits. this multicenter oxidation-reduction enzyme utilizes either nadh or nadph as electron donor and possesses as prosthetic groups two iron-sulfur (fe4s4) clusters, two siroheme groups, and two fad molecules. the native activity of the enzyme is the nad(p)h-dependent reduction of nitrite to ammonia. in addition, n. crassa nitrite reductase di ...19968798648
bioelectrorheological model of the cell. vi. experimental verification of the rheological model of cytoplasmic membrane.the susceptibility of the neurospora crassa (slime) cellular membrane to electroporation and electrodestruction in an alternating electric field was further investigated. the results were analyzed according to the dynamic rheological model of the cytoplasmic membrane. based on an analytical description of membrane susceptibility to electroporation, the rheological parameters of the foregoing process in n. crassa cellular membrane were found: they closely resemble those previously determined for ...19968789120
determination of the neurospora crassa cys 3 sulfur regulatory protein consensus dna-binding site: amino-acid substitutions in the cys3 bzip domain that alter dna-binding specificity.cys3 is the positive-acting global regulatory protein involved in the sulfur control circuit in neurospora crassa and belongs to the family of bzip dna-binding proteins. here we report a characterization of native dna-binding sites recognized by cys3. dna footprinting experiments and systematic mutational analysis were used to define the consensus cys3-binding sequence, 5'-atgpupypupycat, a 10-bp palindrome. the sequence 5'-atgacgtcat acts as a strong binding site, and all single nucleotide chan ...19968781172
cloning and analysis of the alternative oxidase gene of neurospora crassa.mitochondria of neurospora crassa contain a cyanide-resistant alternative respiratory pathway in addition to the cytochrome pathway. the alternative oxidase is present only when electron flow through the cytochrome chain is restricted. both genomic and cdna copies for the alternative oxidase gene have been isolated and analyzed. the sequence of the predicted protein is homologous to that of other species. the mrna for the alternative oxidase is scarce in wild-type cultures grown under normal con ...19968770590
a uv-induced mutation in neurospora that affects translational regulation in response to arginine.the neurospora crassa arg-2 gene encodes the small subunit of arginine-specific carbamoyl phosphate synthetase. the levels of arg-2 mrna and mrna translation are negatively regulated by arginine. an upstream open reading frame (uorf) in the transcript's 5' region has been implicated in arginine-specific control. an arg-2-hph fusion gene encoding hygromycin phosphotransferase conferred arginine-regulated resistance to hygromycin when introduced into n. crassa. we used an arg-2-hph strain to selec ...19968770589
a stringent role of f-actin in the ascogonial differentiation of neurospora crassa fluffy mutant.cytochalasin d, an inhibitor of actin polymerization, interferes with ascogonial differentiation in the fertile fluffy mutant of neurospora crassa. as the total level of actin and its mrna remain unchanged, this suggests that it is in its microfilamentous form (f-form) that actin is stringently required for female differentiation.19958768259
evidence for a novel class of microbial 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase in streptomyces coelicolor a3(2), streptomyces rimosus and neurospora crassa.the tryptophan-sensitive 3-deoxy-d-arabino-heptulosonate-7-phosphate (dahp) synthases from streptomyces coelicolor a3(2), streptomyces rimosus and neurospora crassa have been purified to homogeneity. all three enzymes have a subunit mr of 54,000. the s. coelicolor dahp synthase was physically and kinetically characterized and the n-terminal amino acid sequence was obtained. the n-terminal amino acid sequence could not be obtained for the enzymes from s. rimosus and n. crassa, their n-termini app ...19968760910
indications for the occurrence of nitric oxide synthases in fungi and plants and the involvement in photoconidiation of neurospora crassa.indications for the occurrence of nitric oxide synthases in dictyostelium, neurospora, phycomyces and the leguminous plant mucuna hassjoo as well as a physiological role of nitric oxide in neurospora crassa are demonstrated. an exogenous nitic oxide donor, sodium nitroprusside, inhibited light-stimulated conidiation in n. crassa. specific inhibitors of nitric oxide synthase, like the arginine derivatives ng -nitro-l-arginine (l-na) and ng-nitro-l-arginine-methyl ester (l-name), enhanced conidiat ...19968760579
analysis of the cyt-18 protein binding site at the junction of stacked helices in a group i intron rna by quantitative binding assays and in vitro selection.the neurospora crassa mitochondrial tyrosyl-trna synthetase (cyt-18 protein) functions in splicing group i introns by promoting the formation of the catalytically active structure of the intron rna. previous studies showed that cyt-18 binds with high affinity to the p4-p6 domain of the catalytic core and that there is some additional contribution to binding from the p3-p9 domain. here, quantitative binding assays with deletion derivatives of the n. crassa mitochondrial large rrna intron showed t ...19968760500
the localization of chitin synthase in membranous vesicles (chitosomes) in neurospora crassa.polyclonal anti-chitin synthase antibodies raised against the saccharomyces cerevisiae chs2 gene product were used to identify and localize chitin synthase in the filamentous ascomycete neurospora crassa. a single band of approximately 110 kda was observed in western blots of total protein extracts of n. crassa, probed with these antibodies. however, several additional bands were labelled when membrane fraction proteins (microsomes) were probed. histo-immunochemical localization of chitin syntha ...19968757723
nut1, a major nitrogen regulatory gene in magnaporthe grisea, is dispensable for pathogenicity.nut1, a gene homologous to the major nitrogen regulatory genes nit-2 of neurospora crassa and area of aspergillus nidulans, was isolated from the rice blast fungus, magnaporthe grisea. nut1 encodes a protein of 956 amino acid residues and, like nit-2 and area, has a single putative zinc finger dna-binding domain. functional equivalence of nut1 to area was demonstrated by introducing the nut1 gene by dna-mediated transformation into an area loss-of-function mutant of a. nidulans. the introduced n ...19968757395
downstream 28s gene sequences on the rna template affect the choice of primer and the accuracy of initiation by the r2 reverse transcriptase.r2 non-long terminal repeat retrotransposable elements insert at a unique site in the 28s rrna genes of insects. the protein encoded by the single open reading frame of r2 is capable of conducting the initial steps of its integration in vitro. the protein nicks the noncoding strand of the 28s target dna (the strand which serves as a template for rna synthesis) and uses the 3' hydroxyl group exposed by this nick to prime reverse transcription of the r2 rna template. this target-primed reverse tra ...19968756630
a reverse transcriptase activity in potato mitochondria.a reverse transcriptase activity has been detected in potato mitochondria using special rnas as templates: a bacterial rna coding for neomycin phosphotransferase (neo pa rna) and a neurospora crassa mitochondrial rna (184 nt rna). surprisingly, no exogenous primer addition was required. these rna templates share a primary and secondary structure similar to the t psi cg loop of trnas that could constitute the recognition site for the enzyme. reverse transcriptase activity was inhibited by ddttp, ...19968756599
the isoprenoid pathway: cloning and characterization of fungal fpps genes.farnesylpyrophosphate synthase (fpps) is a key enzyme in isoprenoid biosynthesis. several classes of essential metabolites, including sterols, quinones, carotenoids and gibberellins, are terpenoids with high biological activity. the structural gene for fpp synthase was isolated from two ascomycete fungi, neurospora crassa and gibberella fujikuroi. a comparative analysis of the nucleotide sequences of both fpps genes revealed the presence of introns at the same positions at the 5' end of the codi ...19968753652
the mus-8 gene of neurospora crassa encodes a structural and functional homolog of the rad6 protein of saccharomyces cerevisiae.we cloned a dna repair gene, mus-8, of neurospora crassa and sequenced the genomic dna and cdna. nucleotide-sequence analysis indicated that the mus-8 gene contains an open reading frame (orf) of 456 bp, interrupted by three small introns. the deduced amino-acid sequence showed that the mus-8 gene encodes a 17 kda protein which has 77.5% and 83.3% identity to the rad6 protein of saccharomyces cerevisiae and the rhp6(+) protein of schizosaccharomyces pombe, respectively. the rad6 protein is a ubi ...19968753651
p150glued, the largest subunit of the dynactin complex, is nonessential in neurospora but required for nuclear distribution.dynactin is a multisubunit complex that is required for cytoplasmic dynein, a minus-end-directed, microtubule-associated motor, to efficiently transport vesicles along microtubules in vitro. p150glued, the largest subunit of dynactin, has been identified in vertebrates and drosophila and recently has been shown to interact with cytoplasmic dynein intermediate chains in vitro. the mechanism by which dynactin facilitates cytoplasmic dynein-dependent vesicle transport is unknown. we have devised a ...19968744947
the vacuolar atpase of neurospora crassa is indispensable: inactivation of the vma-1 gene by repeat-induced point mutation.to analyze the phenotype of cells lacking the vacuolar atpase, we inactivated the vma-1 gene, which encodes the catalytic subunit of the enzyme. because preliminary experiments suggested the vma-1 gene was essential, we developed a method of simultaneously inactivating the gene and complementing it with a functional copy. we call this method repeat-induced point mutation (rip) & rescue. two strains, both of which contained an extra copy of the vma-1 gene, were mated. progeny that had inherited a ...19968722770
gene conversion alone accounts for more than 90% of recombination events at the am locus of neurospora crassa.we have used closely flanking molecular markers located approximately 4 kb distal and 6 kb proximal of the am locus to investigate the incidence of crossover events associated with the generation of prototrophic recombinants in a cross heteroallelic am1 am6. ninety-three percent of prototrophs were generated by events that did not recombine the molecular markers, indicating that simple conversion accounts for the formation of most prototrophs and that associated crossovers are much less frequent ...19968722768
a kinase-encoding gene from colletotrichum trifolii complements a colonial growth mutant of neurospora crassa.colletotrichum trifolii is a fungal pathogen which is responsible for anthracnose disease of alfalfa. to initiate research on molecular communication in this fungus, a kinase-encoding gene (tb3) and the corresponding cdna were cloned and sequenced. the deduced amino acid sequence of tb3 closely resembles that of a neurospora crassa serine/threonine protein kinase, cot1, required for hyphal elongation and branching. the c-terminal catalytic domains of tb3 and cot1 are highly conserved but the n-t ...19968709963
role of the n- and c-termini of porin in import into the outer membrane of neurospora mitochondria.the signals for targeting and assembly of porin, a protein of the mitochondrial outer membrane, have not been clearly defined. targeting information has been hypothesized to be contained in the n-terminus, which may form an amphipathic alpha-helix, and in the c-terminal portion of the protein. here, the role of the extreme n- and c-termini of porin from neurospora crassa in its import into the mitochondrial outer membrane was investigated. deletion mutants were constructed which lacked the n-ter ...19968706833
characterization of a vacuolar protease in neurospora crassa and the use of gene riping to generate protease-deficient strains.we have isolated a gene from neurospora crassa that appears to encode a pepstatin-sensitive protease found both in membranes and in soluble contents of vacuoles. the gene contains two introns and encodes a 396-residue protein with a molecular mass of 42,900 da. because of the similarity of the protein to proteinase a in saccharomyces cerevisiae the gene has been named pep-4. strains with mutations in the pep-4 gene were generated in vivo by the gene riping procedure described by selker and garre ...19968702999
primary structure of a ferredoxin-like iron-sulfur subunit of complex i from neurospora crassa.we have isolated cdna clones encoding an iron-sulfur polypeptide subunit of the mitochondrial complex i of neurospora crassa. the fungal cdna library was screened by hybridisation with an heterologous probe from paracoccus denitrificans. the dna sequence of relevant isolates was determined and revealed an open reading frame encoding a precursor protein of 219 amino acid residues. the gene product is a ferredoxin-like protein that contains two cysteine-rich motives that may each bind a tetranucle ...19968695631
meiotic transvection in fungi.the neurospora crassa asm-1+ (ascospore maturation 1) gene encodes an abundant nucleus-localized protein required for formation of female structures and for ascospore maturation. deletion mutants of asm-1+ are "ascus-dominant," i.e., when crossed to wild type, neither asm-1+ nor asm-1 delta spores mature. to explain this behavior, we considered three models: an effect of reduced dosage of the gene product, failure of internuclear communication, and failure of transvection (regulation dependent o ...19968689677
molecular cloning of the acr-2 gene which controls acriflavine sensitivity in neurospora crassa.the acr-2 gene of neurospora crassa was cloned by complementation of the wild-type strain by dna from an acriflavine-resistant strain, acr-2. the transcript of the acr-2 gene is 2.3 kb long and contains two leader open reading frames (orfs) that precede the acr-2 coding region and, if translated, they would generate sequences of 23 and 43 amino acid residues, respectively. the predicted acr-2 protein contains 595 amino acids that include a putative zn(ii)cys6 binuclear domain that is followed by ...19968679704
cloning of the gene encoding the mitochondrial adenine nucleotide carrier of schizosaccharomyces pombe by functional complementation in saccharomyces cerevisiae.we describe the isolation and sequencing of both cdna and genomic clones encoding the mitochondrial adp/atp carrier (anc) of schizosaccharomyces pombe (sp). the cdna clone was isolated from a cdna library of this fission yeast by complementation of a saccharomyces cerevisiae (sc) strain defective in adenine nucleotide carrier. the predicted amino acid (aa) sequence (322 aa) shared similarity with the known anc sequences. it is more closely related to neurospora crassa (nc) anc than to scanc1, 2, ...19968675018
transgene silencing of the al-1 gene in vegetative cells of neurospora is mediated by a cytoplasmic effector and does not depend on dna-dna interactions or dna methylation.the molecular mechanisms involved in transgene-induced gene silencing ('quelling') in neurospora crassa were investigated using the carotenoid biosynthetic gene albino-1 (al-1) as a visual marker. deletion derivatives of the al-1 gene showed that a transgene must contain at least approximately 132 bp of sequences homologous to the transcribed region of the native gene in order to induce quelling. transgenes containing only al-1 promoter sequences do not cause quelling. specific sequences are not ...19968670816
fungal membrane responses induced by plant defensins and thionins.treatment of hyphae of neurospora crassa with antifungal plant defensins, i.e. rs-afp2 and dm-amp1 isolated from radish and dahlia seed, respectively, induced a rapid k+ efflux, ca2+ uptake, and alkalinization of the incubation medium. the rs-afp2-induced alkalinization of the incubation medium could be inhibited with g-protein inhibitors. alpha-hordothionin, an antifungal thionin from barley seed, caused a sustained increased ca2+ uptake at subinhibitory concentrations but only a transient incr ...19968663029
the role of the n and c termini of recombinant neurospora mitochondrial porin in channel formation and voltage-dependent gating.to investigate the role of the n and c termini in channel function and voltage-dependent gating of mitochondrial porin, we expressed wild-type and mutant porins from neurospora crassa as his-tag fusion products in escherichia coli. large quantities of the proteins were purified by chromatography across a nickle-nitrilotriacetic acid-agarose column under denaturing conditions. the purified his-tagged wild-type protein could be functionally reconstituted in the presence of detergent and sterol and ...19968662769
characterization of the aspergillus parasiticus niad and niia gene cluster.the nitrate reductase gene (niad) and nitrite reductase gene (niia) of aspergillus parasiticus are clustered and are divergently transcribed from a 1.6-kb intergenic region (niad-niia). the deduced aminoacid sequence of the a. parasiticus nitrate reductase demonstrated a high degree of homology to those of other aspergillus species, as well as to leptosphaeria maculans, fusarium oxysporum, gibberella fujikuroi and neurospora crassa, particularly in the cofactor-binding domains for molybdenum, he ...19968662212
molecular cloning of thi-4, a gene necessary for the biosynthesis of thiamine in neurospora crassa.the thiamine-4 (thi-4) gene was cloned by functional complementation of a thi-4 mutant of neurospora crassa. the product of this gene is believed to be involved in the condensation of pyrimidine and thiazole precursors, which is necessary for the synthesis of thiamine. the thi-4 gene has ten introns which vary in length from 57 to 200 bp and the junction and internal (lariat) sequences are in good agreement with the consensus sequences for the splicing of introns in n. crassa. the thi-4 gene enc ...19968662211
over-expression of the nud1-coded endo-exonuclease in saccharomyces cerevisiae enhances dna recombination and repair.the nud1(=nuc2) gene of saccharomyces cerevisiae has been subcloned and over-expressed in multi-copy plasmids. enhanced expression of this nuclear endo-exonuclease gene was confirmed by northern hybridization (>10-fold increase), and increased enzymatic activity (2.4-fold increase) was demonstrated by direct immunological assay using antibody raised against the purified neurospora crassa endo-exonuclease. we found that increased expression of nud1 was associated with an increase in cell survival ...19968662209
physical and genetic map of the mitochondrial genome of cryphonectria parasitica ep155.in the chestnut-blight fungus, cryphonectria parasitica, a cytoplasmically transmissible (infectious) form of hypovirulence is associated with mitochondrial dna (mtdna) mutations that cause respiratory deficiencies. to facilitate the characterization of such mutations, a restriction map including the probable location of 13 genes was constructed for a relatively well-characterized virulent strain of the fungus, ep155. the physical map is based on the order of all fragments generated by cleavage ...19968662207
the regulatory protein nit4 that mediates nitrate induction in neurospora crassa contains a complex tripartite activation domain with a novel leucine-rich, acidic motif.expression of nit-3 and nit-6, the structural genes which encode nitrate reductase and nitrite reductase in neurospora crassa, requires the global-acting nit2 and the pathway specific nit4 regulatory proteins. nit4, which consists of 1090 amino-acid residues, possesses a cys6/zn2 zinc cluster dna-binding-domain. nit4 was dissected to localize transactivation domains by fusion of various segments of nit4 to the dna-binding domain of gal4 for in vivo analysis in yeast. three separate activation su ...19968662193
isolation and sequence analysis of the cdna encoding delta 1-pyrroline-5-carboxylate reductase from zalerion arboricola.a cdna encoding delta 1-pyrroline-5-carboxylate reductase (p5cr) was isolated from the pneumocandin (pmo)-producing fungus, zalerion arboricola (za), by complementation of a p5cr-deficient mutant (pro3) of saccharomyces cerevisiae (sc). the cloned cdna was placed under control of the sc galactokinase (gal1) promoter and restored p5cr activity to the pro3 mutant. sequence analysis revealed that the za p5cr-encoding cdna encodes an approx. 35 kda protein with substantial amino acid (aa) identity t ...19968654976
a receptor for the import of proteins into human mitochondria.we have characterised a 16.3-kda human protein that functions as a receptor for the import of preproteins into mitochondria. based on amino acid sequence alignments, the protein (hmas20p) is 41% similar to mas20p (20-kda mitochondrial assembly protein) from yeast saccharomyces cerevisiae and 38% similar to mom19 (19-kda mitochondrial outer-membrane protein) from neurospora crassa. hmas20p has a putative n-terminal transmembrane sequence of 29 amino acids and an acidic c-terminus. a 13-kda fragme ...19968654425
a tyrosyl-trna synthetase protein induces tertiary folding of the group i intron catalytic core.the neurospora crassa mitochondrial tyrosyl-trna synthetase (cyt-18 protein) functions in splicing group i introns. we have used chemical-structure mapping and footprinting to investigate the interaction of the cyt-18 protein with the n. crassa mitochondrial large subunit ribosomal rna (mt lsu) and nd1 introns, which are not detectably self-splicing in vitro. our results show that both these non-self-splicing introns form most of the short range pairings of the conserved group i intron secondary ...19968648621
genetic interactions among cytoplasmic dynein, dynactin, and nuclear distribution mutants of neurospora crassa.cytoplasmic dynein is a multisubunit, microtubule-associated, mechanochemical enzyme that has been identified as a retrograde transporter of various membranous organelles. dynactin, an additional multisubunit complex, is required for efficient dynein-mediated transport of vesicles in vitro. recently, we showed that three genes defined by a group of phenotypically identical mutants of the filamentous fungus neurospora crassa encode proteins that are apparent subunits of either cytoplasmic dynein ...19968643479
signal transduction in pneumocystis carinii: characterization of the genes (pcg1) encoding the alpha subunit of the g protein (pcg1) of pneumocystis carinii carinii and pneumocystis carinii ratti.pneumocystis carinii is a eukaryotic organism that causes pneumonia in immunocompromised hosts. the cell biology and life cycle of the organism are poorly understood primarily because of the lack of a continuous in vitro cultivation system. these limitations have prevented investigation of the organism's infectious cycle and hindered the rational development of new antimicrobial therapies and implementation of measures to prevent exposure to the organism or transmission. the interaction of p. ca ...19968641768
the sequence of a small subunit of cytochrome c oxidase from crithidia fasciculata which is homologous to mammalian subunit iv.the sequence of subunit 8 of cytochrome c oxidase from crithidia fasciculata was determined by sequencing cdna and n-terminus of the mature protein (mr = 15.7 kda). the (inferred) protein is homologous to mammalian cox iv and the corresponding cox subunits from yeast, neurospora crassa and dictyostelium discoideum, which is reflected in a very similar hydropathy profile. elements that are conserved in the c. fasciculata sequence include (i) an n-terminal (d/e)-(k/r)-x-k-(x2)-w-(x2)-(i/l) motif, ...19968641419
insights into the molecular basis of thermal stability from the structure determination of pyrococcus furiosus glutamate dehydrogenase.the structure determination of the glutamate dehydrogenase from the hyperthermophile pyrococcus furiosus has been completed at 2.2 a resolution. the structure has been compared with the glutamate dehydrogenases from the mesophiles clostridium symbiosum, escherichia coli and neurospora crassa. this comparison has revealed that the hyperthermophilic enzyme contains a striking series of networks of ion-pairs which are formed by regions of the protein which contain a high density of charged residues ...19968639325
role of an upstream open reading frame in mediating arginine-specific translational control in neurospora crassa.the neurospora crassa arg-2 transcript contains an upstream open reading frame (uorf) specifying a 24-residue leader peptide and is subject to a novel form of negative translational regulation in response to arginine. the role of the arg-2 uorf in arginine-specific negative regulation was investigated by using translational fusions of wild-type and mutant arg-2 sequences to the escherichia coli lacz reporter gene specifying beta-galactosidase. the wild-type uorf conferred arg-specific regulation ...19968636015
characterization of the biophysical and motility properties of kinesin from the fungus neurospora crassa.neurospora kinesin (nkin) is a distant relative of the family of conventional kinesins, members of which have been identified in various animal species. as in its animal counterparts, nkin most likely is an organelle motor. because it is a functional homologue of the kinesin heavy chain of higher eukaryotes, its biophysical and motility properties were compared with those of other conventional kinesins. purified nkin behaves as a homodimeric complex composed of two subunits of a 105-kda polypept ...19968631782
transcriptional analysis of the mta idiomorph of neurospora crassa identifies two genes in addition to mta-1.in neurospora crassa, mating and heterokaryon formation between opposite mating-types is controlled by a single locus with two alternate forms termed mt a and mt a. previously, an open reading frame (mt a-1) that confers mating identity and heterokaryon incompatibility was characterized in the 5.3 kb mt a idiomorph. in this study, we describe the structural and transcriptional characterization of two additional genes in the mt a idiomorph, mt a-2 and mt a-3. the 373 amino acid mt a-2 orf has 23% ...19968628238
chs-4, a class iv chitin synthase gene from neurospora crassa.in saccharomyces cerevisiae, most of the cellular chitin is produced by chitin synthase iii, which requires the product encoded by the csd2/cal1/dit101/kt12 gene. we have identified, isolated and structurally characterized as csd2/cal1/dit101/kt12 homologue in the filamentous ascomycete neurospora crassa and have used a "reverse genetics" approach to determine its role in vivo. the yeast gene was used as a heterologous probe for the isolation of a n. crassa gene(designated chs-4) encoding a poly ...19968628221
cloning and characterization of the neurospora crassa cyt-5 gene. a nuclear-coded mitochondrial rna polymerase with a polyglutamine repeat.the neurospora crassa mutants, cyt-5-1 and cyt-5-4 have a cytochrome b- and aa3-deficient phenotype, suggesting that they result from a deficiency in a nuclear-coded component of the mitochondrial gene expression apparatus (bertrand, h., nargang, f. e., colllins, r. a., and zagozeski, c. a. (1977) mol. gen. genet. 153,247-257). the complementing wild-type gene has been cloned and and shown to encode a protein with significant sequence similarity to saccharomyces cerevisiae mitochondrial rna poly ...19968626458
hyphal development in neurospora crassa: involvement of a two-component histidine kinase.two-component signal transduction systems are most often found in prokaryotic organisms where they are responsible for mediating the cellular responses to many environmental stimuli. these systems are composed of an autophosphorylating histidine kinase and a response regulator. we have found evidence for the existence of two-component histidine kinases in the eukaryotic filamentous fungus neurospora crassa based on screening with degenerate primers to conserved regions of these signaling protein ...19968622950
cloning and sequence of the ascobolus immersus s-adenosyl-l-methionine synthetase-encoding gene.the structural gene encoding s-adenosyl-l-methionine synthetase (sam-s) in the fungus ascobolus immersus has been cloned and sequenced. it contains a 1179-bp orf, interrupted by three introns, encoding a 393-amino-acid protein (42 978 da) that is 90% homologous to the sam-s of the filamentous fungus neurospora crassa, indicating that these fungi are closely related species.19968621082
an additional copy of the adenylate cyclase-encoding gene relieves developmental defects produced by a mutation in a vegetative incompatibility-controlling gene in podospora anserina.to identify cellular functions involved in vegetative incompatibility in filamentous fungi, we have initiated the cloning of podospora anserina (pa) mod genes. these genes interfere with the lethal reaction triggered by interaction between incompatible het genes. a gene (pa ac) has been cloned by complementation of developmental defects caused by a mutation in the mod-d gene. this gene encodes a protein of 2145 amino acids (aa)that exhibits strong similarities with many adenylate cyclases (ac). ...19968621071
transient expression of a mitochondrial precursor protein. a new approach to study mitochondrial protein import in cells of higher eukaryotes.in order to study mitochondrial protein import in the context of whole cell metabolism, we have used the transfection technique based on semliki forest virus (sfv) to express a mitochondrial precursor protein within bhk21 cells and human fibroblasts. recombinant sfv particles mediate a highly efficient, transient transfection of higher eukaryotic cells. the mitochondrial precursor protein used is a fusion protein consisting of the mitochondrial targeting sequence of neurospora crassa atpase subu ...19968620886
purification, kinetic characterization and involvement of tryptophan residue at the nadph binding site of xylose reductase from neurospora crassa.xylose reductase (xr) from neurospora crassa was purified to homogeneity and was found to be specific to nadph (nicotinamide adenine dinucleotide phosphate). the purified enzyme showed m(r) of 60 and 29 kda by gel filtration and sds-page indicating the presence of two subunits. the kinetic mechanism of xylose reductase is 'iso-ordered bi bi'. inactivation of xr by n-bromosuccinimide (nbs) was found to be biphasic with second-order rate constants of 2.5 x 10(2) and 80 m-1s-1 for the fast (kf) and ...19968620033
molecular cloning and functional expression of human deoxyhypusine synthase cdna based on expressed sequence tag information.deoxyhypusine synthase is an nad(+)-dependent enzyme that catalyses the formation of a deoxyhypusine residue on the eukaryotic initiation factor 5a (eif-5a) precursor by transferring an aminobutyl moiety from spermidine to the epsilon-amino group of a unique lysine residue. we have recently cloned and characterized the neurospora crassa deoxyhypusine synthase cdna using a reverse genetics approach. a genbank search showed that a stretch of the deduced amino acid sequence (96 amino acids) of neur ...19968615810
characterization of a uv endonuclease gene from the fission yeast schizosaccharomyces pombe and its bacterial homolog.from the fission yeast schizosaccharomyces pombe, a cdna fragment was isolated, which confers uv resistance on repair deficient escherichia coli host cells. the cloned cdna encodes a protein of 68,815 da, which has a 36.6% identity of amino acid sequence with the previously identified 74 kda uv endonuclease of the filamentous fungus neurospora crassa. analysis of several truncated gene constructs shows that only the c-terminal two thirds region, which has 54% identity of amino acid sequence with ...19968614629
white collar-1, a central regulator of blue light responses in neurospora, is a zinc finger protein.the neurospora crassa blind mutant white collar-1 (wc-1) is pleiotropically defective in all blue light-induced phenomena, establishing a role for the wc-1 gene product in the signal transduction pathway. we report the cloning of the wc-1 gene isolated by chromosome walking and mutant complementation. the elucidation of the wc-1 gene product provides a key piece of the blue light signal transduction puzzle. the wc-1 gene encodes a 125 kda protein whose encoded motifs include a single class four, ...19968612589
isolation of the vma-6 gene encoding a 41 kda subunit of the neurospora crassa vacuolar atpase, and an adjoining gene encoding a ribosome-associated protein.the vma-6 gene, encoding a membrane-associated subunit of the vacuolar h+-atpase from neurospora crassa, was cloned and sequenced. the gene contains three small introns and encodes a protein of 41 005 da. when compared with homologous polypeptides from other species, the n. crassa protein contains a unique glycine-rich region. three conserved cysteine residues, previously unrecognized, have been identified. an unrelated gene encoding a protein of 31 701 da was found 2.1 kb downstream of vma-6. t ...19968611592
distribution of thiols in microorganisms: mycothiol is a major thiol in most actinomycetes.mycothiol [2-(n-acetylcysteinyl)amido-2-deoxy-alpha-d-glucopyranosyl- (1-->1)-myo-inositol] (msh) has recently been identified as a major thiol in a number of actinomycetes (s. sakuda, z.-y. zhou, and y. yamada, biosci. biotech. biochem. 58:1347-1348, 1994; h. s. c. spies and d. j. steenkamp, eur. j. biochem. 224:203-213, 1994; and g. l. newton, c. a. bewley, t. j. dwyer, r. horn, y. aharonowitz, g. cohen, j. davies, d. j. faulkner, and r. c. fahey, eur. j. biochem. 230:821-825, 1995). since thi ...19968606174
gel mobility shift scanning of the acetate-inducible promoters from neurospora crassa reveals a common co-inducible dna-binding protein.the promoter regions of four acetate-inducible genes of neurospora crassa, acu-3, acu-5, acu-8 and acu-9, have been sequenced. using a scanning gel mobility shift assay particular dna regions in each promoter have been shown specifically to bind partially purified protein extracted from acetate-induced mycelia. the protein-binding regions so defined have common sequence motifs, elements of which are similar to those required for acetate induction in aspergillus nidulans.19968602159
nucleotide sequence and expression of the gene encoding nadp+- dependent glutamate dehydrogenase (gdha) from agaricus bisporus.the gene encoding nadp+-dependent glutamate dehydrogenase (gdha) was isolated from an agaricus bisporus recombinant phage lambda library. the deduced amino acid sequence would specify a 457-amino acid protein that is highly homologous in sequence to those derived from previously isolated and characterized genes coding for microbial nadp+-gdh. the open reading frame is interrupted by six introns. none of the introns is located at either one of the positions of the two introns conserved in the cor ...19968602149
nutritional and growth control of ribosomal protein mrna and rrna in neurospora crassa.the effects of changing growth rates on the levels of 40s pre-rrna and two r-protein mrnas were examined to gain insight into the coordinate transcriptional regulation of ribosomal genes in the ascomycete fungus neurospora crassa. growth rates were varied either by altering carbon nutritional conditions, or by subjecting the isolates to inositol-limiting conditions. during carbon up- or down-shifts, r-protein mrna levels were stoichiometrically coordinated. changes in 40s pre-rrna levels paralle ...19968600464
mutational analysis reveals dispensability of the n-terminal region of the aspergillus transcription factor mediating nitrogen metabolite repression.mutational analysis has enabled identification and localization of an upstream exon of the area gene of aspergillus nidulans mediating nitrogen metabolite repression. a mutation in the initiation codon and frameshift mutations, which revert by restoration of the reading frame, established the coding role of the exon and mutations affecting intron splicing in conjunction with dna sequencing of reverse transcriptase polymerase chain reaction (rt-pcr) products localized the coding region intron. th ...19958596437
functional molecular aspects of the nadh dehydrogenases of plant mitochondria.there are multiple routes of nad(p)h oxidation associated with the inner membrane of plant mitochondria. these are the phosphorylating nadh dehydrogenase, otherwise known as complex i, and at least four other nonphosphorylating nad(p)h dehydrogenases. complex i has been isolated from beetroot, broad bean, and potato mitochondria. it has at least 32 polypeptides associated with it, contains fmn as its prosthetic group, and the purified enzyme is sensitive to inhibition by rotenone. in terms of su ...19958595975
mapping translocation breakpoints by orthogonal field agarose-gel electrophoresis.orthogonal field agarose-gel electrophoresis (ofage) of chromosomes from translocation-bearing and normal neurospora crassa strains was utilized, first, to recover cosmids from a translocated region, and second, to map translocation breakpoints. surprisingly, the right breakpoints in two independently derived, interstitial translocations, t(ii-->iii) ar18 and t(ii-->vi)p2869, are within about 5.6 kbp of each other suggesting that this region of linkage group (lg) ii may be fragile or otherwise s ...19968595678
the orientation of gene maps by recombination of flanking markers for the am locus of neurospora crassa.fincham (1967), smyth (1973 b) and rambosek and kinsey (1983) have each generated fine-structure maps of the am gene of neurospora crassa. each map had a consistent linear order of alleles but the assignment of an orientation with respect to other linkage group-v loci differed. fincham found the end marked by the am6 allele to be at the distal end of the locus, smyth found am6 to be at the proximal end while the data of rambosek and kinsey did not suggest an orientation. smyth s orientation has ...19958595655
sequence repeat-induced disruption of the major heat-inducible hsp70 gene of neurospora crassa.the process of repeat-induced point mutation (rip) was used to disrupt hsps-1, the gene encoding the major heat-inducible member of the hsp70 family of neurospora crassa. a plasmid dna, containing an incomplete copy of hsps-1 and the selectable marker qa-2+, was introduced into germinated conidia. the sexual progeny of transformants with ectopically integrated hsps-1 dna was examined for rip by southern-blot analysis of mboi- and sau3a-digested genomic dna. progeny strains, showing rip, were tes ...19958595654
different respiratory-defective phenotypes of neurospora crassa and saccharomyces cerevisiae after inactivation of the gene encoding the mitochondrial acyl carrier protein.the nuclear genes (acp-1, acp1) encoding the mitochondrial acyl carrier protein were disrupted in neurospora crassa and saccharomyces cerevisiae. in n. crassa acp-1 is a peripheral subunit of the respiratory nadh : ubiquinone oxidoreductase (complex i). s. cerevisiae lacks complex i and its acp1 appears to be located in the mitochondrial matrix. the loss of acp-1 in n. crassa causes two biochemical lesions. firstly, the peripheral part of complex i is not assembled, and the membrane part is not ...19958595652
ubiquinol-cytochrome-c oxidoreductase of neurospora crassa. 19958592472
generation and characterization of nadh: ubiquinone oxidoreductase mutants in neurospora crassa. 19958592454
mitochondrial receptor complex from neurospora crassa and saccharomyces cerevisiae. 19958592451
expression of the neurospora crassa metallothionein gene in escherichia coli and its effect on heavy-metal uptake.the gene coding for the neurospora crassa metallothionein protein was chemically synthesized and cloned into the fusion expression vectors pmal-c and pmal-p. cell-fractionation experiments demonstrated the proper localization of the pmal-c and pmal-p- expressed proteins to the cytosol and periplasm of the bacteria respectively. control bacteria as well as the recombinant bacteria producing the metallothionein protein were incubated with solutions of 109cd at concentrations of 0.2 microm, 1 micro ...19958590662
the chromosomal region which includes the recombinator cog in neurospora crassa is highly polymorphic.the st lawrence st74-or23-iva and lindegren y8743 strains of neurospora crassa have a different provenance from wild collections and dissimilar cog alleles; that in lindegren, cogla (previously designated cog+), is a more efficient recombinator than cogs74a and cogea (previously cog), the alleles in st lawrence and emerson a respectively. restriction fragment length polymorphisms (rflps) and sequence polymorphisms (sps) were used to map the difference between cogla and cogs74a to a region that e ...19958590467
the novel nuclear gene dss-1 of saccharomyces cerevisiae is necessary for mitochondrial biogenesis.a previously unknown nuclear gene dss-1 from saccharomyces cerevisiae was cloned and sequenced. the gene was isolated as a multicopy suppressor of a disruption of the suv-3 gene coding for a dead/h box protein involved in processing and turnover of mitochondrial transcripts. the dss-1 gene codes for a 970 amino-acid protein of molecular weight 111 kda and is necessary for mitochondrial biogenesis. amino-acid sequence analysis indicates the presence of motifs characteristic for escherichia coli r ...19958590460
the neurospora organelle motor: a distant relative of conventional kinesin with unconventional properties.the "conventional" kinesins comprise a conserved family of molecular motors for organelle transport that have been identified in various animal species. organelle motors from other phyla have not yet been analyzed at the molecular level. here we report the identification, biochemical and immunological characterization, and molecular cloning of a cytoplasmic motor in a "lower" eukaryote, the ascomycete fungus neurospora crassa. this motor, termed nkin (for neurospora kinesin), exhibits several un ...19958589459
conidial alkaline phosphatase from neurospora crassa.an alkaline phosphatase was purified from conidia of a neurospora crassa wild type strain. the m(r) of the purified native enzyme was estimated as ca 145,000 and 110,000 by gel filtration, in the presence and absence of magnesium ions, respectively. a single polypeptide band of m(r) 36,000 was detected by sds-page, suggesting that the native enzyme was a tetramer of apparently identical subunits. conidial alkaline phosphatase was an acidic protein (pl = 4.0 +/- 0.1), with 40% carbohydrate conten ...19968588877
alterations in growth rate associated with a normally persisting circadian rhythm during spaceflight.biological clocks time many physiological parameters with periodicities close to 24 h; those which persist in the absence of environmental cues are circadian. an earlier shuttle experiment (sts-9) examined circadian pacemaker function and growth rate of neurospora crassa and demonstrated damped rhythm amplitudes, increased variability in period lengths and altered growth rates.19958588798
the roles of ca2+ and plasma membrane ion channels in hyphal tip growth of neurospora crassa.growing hyphae of the ascomycete fungus neurospora crassa contained a tip-high gradient of cytoplasmic ca2+, which was absent in non-growing hyphae and was insensitive to gd3+ in the medium. patch clamp recordings in the cell-attached mode, from the plasma membrane of these hyphae, showed two types of channel activities; spontaneous and stretch activated. the spontaneous channels were identified as inward k+ channels based on inhibition by tetraethylammonium. the stretch activated channels had i ...19958586653
chromosome rearrangements that involve the nucleolus organizer region in neurospora.in approximately 3% of neurospora crassa rearrangements, part of a chromosome arm becomes attached to the nucleolus organizer region (nor) at one end of chromosome 2 (linkage group v). investigations with one inversion and nine translocations of this type are reported here. they appear genetically to be nonreciprocal and terminal. when a rearrangement is heterozygous, about one-third of viable progeny are segmental aneuploids with the translocated segment present in two copies, one in normal pos ...19958582636
inactivation of a single-2a phosphoprotein phosphatase is lethal in neurospora crassa.a pcr approach, employing the use of degenerate oligonucleotide mixtures, was used to isolate pph-1, a type-2a protein phosphatase (catalytic subunit)-encoding gene, from neurospora crassa. the isolated single copy gene is 1327 nucleotides in length, contains four putative introns and encodes a 310 amino-acid polypeptide. pph-1 is located between pdx-1 and col-4 on the right arm of n.crassa linkage group iv. pph-1 transcript levels are highest during the first hours of conidial germination. fail ...19958575020
a suppressor mutation which suppresses adenylyl cyclase mutations in neurospora crassa.a spontaneous suppressor mutant, hah, which suppressed the colonial growth of adenylyl cyclase mutants (cr-1) was isolated. the morphology of cr-1 hah was filamentous, but slightly different from that of wild type on solid medium. the hah strain formed many high aerial hyphae, but did not form any conidia. the expression levels of an adenylyl cyclase gene, nac, in both hah and cr-1 hah were much higher than those in wild type or in cr-1. the level of camp in cr-1 was very low but returned to clo ...19958574907
osmotic effects on the polyamine pathway of neurospora crassa.in bacteria, mammals, and certain plants, the induction of the polyamine synthetic enzyme, ornithine decarboxylase (odc), and the accumulation of its product, putrescine, follows osmotic manipulations of cells. in at least some of these cases, this response is indispensable for survival. we wished to determine whether the polyamine pathway of neurospora crassa was regulated in response to hyper- or hypoosmotic conditions. unlike odc of most other classes of organisms, the n. crassa enzyme and th ...19958574906
the molecular nature of mutations in the mt a-1 gene of the neurospora crassa a idiomorph and their relation to mating-type function.the 293-amino acid mt a-1 orf of the a mating-type idiomorph of neurospora crassa is multifunctional. it confers a mating identity and is responsible for heterokaryon incompatibility. the goal of this study was to dissect the functional regions of mt a-1. new mutants of mt a-1 selected for loss of the incompatibility function were obtained. one new mutant, a(m)99, was partially fertile as a maternal parent. this is the first time that fertility and incompatibility functions have been separated f ...19968569681
structural analysis of mitochondrial dna molecules from fungi and plants using moving pictures and pulsed-field gel electrophoresis.the size and structure of mitochondrial dna (mtdna) molecules was investigated by conventional and pulsed-field gel electrophoresis (pfge) and by analyzing moving pictures during electrophoresis of individual fluorescently labelled mtdna molecules. little or no mtdna that migrated into the gel was found in circular form for fungi (schizosaccharomyces pombe, saccharomyces cerevisiae and neurospora crassa) or plants (brassica hirta, tobacco, voodoo lily and maize). most mtdna migrated as a smear o ...19968568898
cytogenetics of an intrachromosomal transposition in neurospora.knowledge of intrachromosomal transpositions has until now been primarily cytological and has been limited to drosophila and to humans, in both of which segmental shifts can be recognized by altered banding patterns. there has been little genetic information. in this study, we describe the genetic and cytogenetic properties of a transposition in neurospora crassa. in tp(ir-->il)t54m94, a 20 map unit segment of linkage group i has been excised from its normal position and inserted near the centro ...19958565702
distinct cis-acting elements mediate clock, light, and developmental regulation of the neurospora crassa eas (ccg-2) gene.the neurospora crassa eas (ccg-2) gene, which encodes a fungal hydrophobin, is transcriptionally regulated by the circadian clock. in addition, eas (ccg-2) is positively regulated by light and transcripts accumulate during asexual development. to sort out the basis of this complex regulation, deletion analyses of the eas (ccg-2) promoter were carried out to localize the cis-acting elements mediating clock, light, and developmental control. the primary sequence determinants of a positive activati ...19968552078
functional analysis by site-directed mutagenesis of individual amino acid residues in the flavin domain of neurospora crassa nitrate reductase.nitrate reductase of neurospora crassa is a complex multi-redox protein composed of two identical subunits, each of which contains three distinct domains, an amino-terminal domain that contains a molybdopterin cofactor, a central heme-containing domain, and a carboxy-terminal domain which binds a flavin and a pyridine nucleotide cofactor. the flavin domain of nitrate reductase appears to have structural and functional similarity to ferredoxin nadph reductase (fnr). using the crystal structure of ...19958552051
identification and expression of the neurospora crassa mei-3 gene which encodes a protein homologous to rad51 of saccharomyces cerevisiae.the mei-3 gene of neurospora crassa encodes a homolog of the escherichia coli reca and saccharomyces cerevisiae rad51 proteins, which are required for recombination and repair of dna double-strand breaks. to determine the molecular function of mei3 protein, anti-mei3 antibody was prepared and used in western blot analysis. the antibody cross-reacted only with crude extracts prepared from perithecia, the fruiting bodies of neurospora. the molecular weight of the mei3 protein was estimated to be 3 ...19958552049
site-directed mutagenesis of the cysteine residues in the neurospora crassa plasma membrane h(+)-atpase.a high-yield yeast expression system for site-directed mutagenesis of the neurospora crassa plasma membrane h(+)-atpase has recently been reported (mahanty, s. k., rao, u. s., nicholas, r. a., and scarborough, g. a. (1994) j. biol. chem. 269, 17705-17712). using this system, each of the eight cysteine residues in the atpase was changed to a serine or an alanine residue, producing strains c148s and c148a, c376s and c376a, c409s and c409a, c472s and c472a, c532s and c532a, c545s and c545a, c840s a ...19968550588
effects of temperature shifts on the activities of neurospora crassa glycogen synthase, glycogen phosphorylase and trehalose-6-phosphate synthase.conidiospore germlings of neurospora crassa submitted to a heat shock at 45 degrees c accumulate trehalose and degrade glycogen. the opposite occurs upon reincubation at a physiologic temperature (30 degrees c). these observations suggest a temperature-dependent mechanism for the preferential synthesis of one or the other sugar reserve. here we show that concomitant with these shifts of temperature, occurred reversible changes in the activities of glycogen synthase and phosphorylase. glycogen sy ...19968549797
detection of additional restriction fragment length polymorphisms among the weakly virulent (nonaggressive) and highly virulent (aggressive) isolates of leptosphaeria maculans.isolates of leptosphaeria maculans were analyzed for their genetic relatedness based on dna restriction fragment length polymorphisms (rflps), employing as southern hybridization probes a combination of heat shock responsive genes (hsp70 and hsp80 from neurospora crassa), the cutinase gene of magnaporthe grisea, and cloned genomic dna sequences from a virulent strain. southern hybridization analysis revealed a high frequency of dna polymorphism. restriction fragments generated by each enzyme-pro ...19958542555
effects of temperature shifts on the metabolism of trehalose in neurospora crassa wild type and a trehalase-deficient (tre) mutant. evidence against the participation of periplasmic trehalase in the catabolism of intracellular trehalose.the effects of temperature shifts on the metabolism of trehalose in neurospora crassa were studied in conidiospore germlings of a wild type strain, and of a mutant (tre), deficient in the activity of periplasmic trehalase. when the temperature of the medium was raised from 30 degrees c to 45 degrees c both strains accumulated trehalose, either in media supplemented with glucose or with glycerol as carbon sources. the profiles of glycolysis metabolites suggested that at 45 degrees c glycolysis wa ...19958541310
species-specific and mating type-specific dna regions adjacent to mating type idiomorphs in the genus neurospora.mating type idiomorphs control mating and subsequent sexual development in neurospora crassa and were previously shown to be well conserved in other neurospora species. the centromere-proximal flanks of the a and a idiomorphs, but not the distal flanks from representative heterothallic, pseudohomothallic, and homothallic neurospora species contain apparent species-specific and/or mating type-specific sequences adjacent to the well-conserved idiomorphs. the variable flank is bordered by regions t ...19958536961
mutations affecting the biosynthesis of s-adenosylmethionine cause reduction of dna methylation in neurospora crassa.a temperature-sensitive methionine auxotroph of neurospora crassa was found in a collection of conditional mutants and shown to be deficient in dna methylation when grown under semipermissive conditions. the defective gene was identified as met-3, which encodes cystathionine-gamma-synthase. we explored the possibility that the methylation defect results from deficiency of s-adenosylmethionine (sam), the presumptive methyl group donor. methionine starvation of mutants from each of nine complement ...19958532524
isolation and characterization of the catalytic subunit of protein phosphatase 2a from neurospora crassa.the catalytic subunit of protein phosphatase 2a (pp2ac) was purified from neurospora crassa extract by (nh4)2so4-ethanol precipitation followed by deae-sephacel, heparin-sepharose, and monoq chromatography steps about 900-fold to a specific activity of 1200 u/g with a 2% yield. the apparent m(r) of pp2ac was estimated to be 35 kda by gel filtration and 33 kda by sds polyacrylamide gel electrophoresis. half maximal inhibition of pp2ac was achieved at 0.3 nm okadaic acid, 0.1 nm microcystin-lr, 56 ...19958529028
phylogenetic analysis of ten black yeast species using nuclear small subunit rrna gene sequences.the nuclear small subunit rrna genes of authentic strains of the black yeasts exophiala dermatitidis, wangiella dermatitidis, sarcinomyces phaemuriformis, capronia mansonii, nadsoniella nigra var. hesuelica, phaeoannellomyces elegans, phaeococcomyces exophialae, exophiala jeanselmei var. jeanselmei and e. castellanii were amplified by pcr and directly sequenced. a putative secondary structure of the nuclear small subunit rrna of exophiala dermatitidis was predicted from the sequence data. alignm ...19958526477
developmental regulation of the gene for formate dehydrogenase in neurospora crassa.we have isolated and characterized a gene, fdh, from neurospora crassa which is developmentally regulated and which produces formate dehydrogenase activity when expressed in escherichia coli. the gene is closely linked (less than 0.6 kb apart) to the leu-5 gene encoding mitochondrial leucyl-trna synthetase; the two genes are transcribed convergently from opposite strands. the expression patterns of these genes differ: fdh mrna is found only during conidiation and early germination and is not det ...19938509325
the beta subunit of the mitochondrial processing peptidase from rat liver: cloning and sequencing of a cdna and comparison with a proposed family of metallopeptidases.most nuclearly encoded mitochondrial proteins are synthesized with amino-terminal leader peptides that are removed by the mitochondrial processing peptidase (mpp) after translocation. earlier we reported cloning and sequencing of a cdna for the larger subunit (mpp alpha subunit) of this enzyme from rat liver mitochondria. we have now completed the cloning and sequencing of a cdna encoding the smaller subunit of the enzyme (mpp beta subunit) from the same source. the cdna consists of 1570 bp: 17 ...19938506385
cloning and sequencing of the genes coding for the a and b subunits of vacuolar-type na(+)-atpase from enterococcus hirae. coexistence of vacuolar- and f0f1-type atpases in one bacterial cell.the eubacterium enterococcus hirae atcc 9790 possesses a h(+)-translocating atpase, and the deduced amino acid sequences of the genes coding for this enzyme have indicated that it is a typical f0f1-type atpase (shibata, c., ehara, t., tomura, k., igarashi, k., and kobayashi, h. (1992) j. bacteriol. 174, 6117-6124). we cloned the ntpa and ntpb genes coding for the a and b subunits, respectively, of na(+)-translocating atpase from the same bacterium, and the full amino acid sequences of the two su ...19938505293
molecular characterization of conventional and new repeat-induced mutants of nit-3, the structural gene that encodes nitrate reductase in neurospora crassa.nitrate reductase of neurospora crassa is a dimeric protein composed of two identical subunits, each possessing three separate domains, with flavin, heme, and molybdenum-containing cofactors. a number of mutants of nit-3, the structural gene that encodes neurospora nitrate reductase, have been characterized at the molecular level. amber nonsense mutants of nit-3 were found to possess a truncated protein detected by a specific antibody, whereas ssu-1-suppressed nonsense mutants showed restoration ...19938479443
cloning and analysis of the mating type genes from cochliobolus heterostrophus.cochliobolus heterostrophus, a heterothallic ascomycete, has a single mating type locus with two alternate forms called mat-1 and mat-2. mat-1 was cloned by complementing a mat-2 strain using a cosmid library from a mat-1 strain and screening for a homothallic transformant. the cosmid recovered from this transformant was able to re-transform a mat-2 strain to homothallism and mat identity was proven by restriction fragment length polymorphism and conventional genetic mapping. all homothallic tra ...19938479433
deletion mutations in the speed operon: spermidine is not essential for the growth of escherichia coli.null mutants of escherichia coli were constructed that cannot synthesize spermidine, because of deletions in the gene encoding s-adenosylmethionine decarboxylase. these mutants are still able to grow at near normal rates in purified media deficient in polyamines. these results in e. coli differ from recent findings that null mutants of saccharomyces cerevisiae and of neurospora crassa have an absolute growth requirement for spermidine.19938472951
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