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disruption of the nuclear gene encoding the 20.8-kda subunit of nadh: ubiquinone reductase of neurospora mitochondria.the nuclear gene coding for the 20.8-kda subunit of the membrane arm of respiratory chain nadh: ubiquinone reductase (complex i) from neurospora crassa, nuo-20.8, was localized on linkage group i of the fungal genome. a genomic dna fragment containing this gene was cloned and a duplication was created in a strain of n. crassa by transformation. to generate rip (repeat-induced point) mutations in the duplicated sequence, the transformant was crossed with another strain carrying an auxotrophic mar ...19968804391
evidences for adenine nucleotide binding in the subunits of neurospora mitochondrial processing peptidase.the mitochondrial processing peptidase (mpp) of neurospora crassa consists of two subunits termed alpha-and beta-mpp. here we present spectroscopic and chromatographic studies indicative of adenine nucleotide binding in the two mpp subunits. adp was identified as the cofactor of alpha-mpp and atp as the cofactor of beta-mpp. the nucleotides are not covalently bound and exert strong control on the conformational and functional properties of the subunits. the adp cofactor of alpha-mpp seems to be ...19968806742
catalytic mechanism of mitochondrial processing peptidase: fluorescence studies.processing of nuclear-encoded precursor proteins by mitochondrial processing peptidase (mpp) is an essential step for their sorting and function in mitochondria. we report spectroscopic studies on the catalytic mechanism of neurospora crassa mpp. it is a complex enzyme consisting of two different subunits termed alpha-and beta-mpp. following changes in the protein intrinsic fluorescence we register and characterize a complex formation between (i) the alpha- and the beta-subunit of mpp, (ii) the ...19968806741
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 g alpha i homologue gna-1 controls multiple differentiation pathways in neurospora crassa.heterotrimeric g proteins are components of principal signaling pathways in eukaryotes. in higher organisms, alpha subunits of g proteins have been divided into four families, gi, gs, gq, and g12. we previously identified a g alpha i homologue gna-1 in the filamentous fungus neurospora crassa. now we report that deletion of gna-1 leads to multiple phenotypes during the vegetative and sexual cycles in n. crassa. on solid medium, delta gna-1 strains have a slower rate of hyphal apical extension th ...19968856670
the product of the het-c heterokaryon incompatibility gene of neurospora crassa has characteristics of a glycine-rich cell wall protein.filamentous fungi are capable of hyphal fusion, but heterokaryon formation between different isolates is controlled by specific loci termed het loci. heterokaryotic cells formed between strains of different het genotype are rapidly destroyed or strongly inhibited in their growth. in neurospora crassa, at least 11 loci, including the mating type locus, affect the capacity to form a heterokaryon between different isolates. in this report, we describe the molecular characterization of the vegetativ ...19968844148
circular dichroism studies of the mitochondrial channel, vdac, from neurospora crassa.the protein that forms the voltage-gated channel vdac (or mitochondrial porin) has been purified from neurospora crassa. at room temperature and ph 7, the circular dichoism (cd) spectrum of vdac suspended in octyl beta-glucoside is similar to those of bacterial porins, consistent with a high beta-sheet content. when vdac is reconstituted into phospholipid liposomes at ph 7, a similar cd spectrum is obtained and the liposomes are rendered permeable to sucrose. heating vdac in octyl beta-glucoside ...19968842216
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
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
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
isolation and characterization of a copper containing protein from blue cell walls of neurospora crassa.culturing neurospora crassa in presence of toxic amounts of copper (0.63 mm) resulted in blue coloured mycelia and cell walls. significant amounts (approximately 45%) of total mycelial copper were associated with cell wall isolates under conditions of copper toxicity. hence, such blue cell walls were analysed to identify specific ligands involved in copper binding. while decuprification of the blue cell walls with 8-hydroxy quinoline (8 hq) did not alter their copper binding abilities, similar t ...19968979504
purification and characterization of cytochrome c oxidase from the insect trypanosomatid crithidia fasciculata.cytochrome c oxidase was purified from the mitochondrial lysate of the insect trypanosomatid crithidia fasciculata with the aid of a methyl hydrophobic interaction column in a rapid one-step procedure. the purified complex displayed all characteristics expected from a eukaryotic cytochrome c oxidase: the presence of cua in electron paramagnetic resonance analysis, a characteristic 605 nm peak in reduced-minus-oxidized optical spectroscopy, and the capacity to efficiently oxidize homologous, but ...19968844671
mutants with altered sensitivity to a calmodulin antagonist affect the circadian clock in neurospora crassa.two newly isolated mutant strains of neurospora crassa, cpz-1 and cpz-2, were hypersensitive to chlorpromazine with respect to mycelial growth but responded differently to the drug with respect to the circadian conidiation rhythm. in the wild type, chlorpromazine caused shortening of the period length of the conidiation rhythm. pulse treatment with the drug shifted the phase and inhibited light-induced phase shifting in neurospora. by contrast to the wild type, the cpz-2 strain was resistant to ...19968807291
isolation and analysis of the arg-13 gene of neurospora crassa.mutations in arg-13 result in slow growth in minimal medium and can suppress mutations in carbamyl phosphate synthase-aspartate carbamyl transferase within the pyrimidine pathway; the exact biochemical function of the gene product is unknown. to understand the role of arg-13 in arginine metabolism, cosmids rescuing growth in arg-13 mutants were cloned and mapped to the position of arg-13 on lg ir. northern analysis showed the arg-13 message to contain approximately 2100 nt, although a 1.4-kb gen ...19968807290
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
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
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
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
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
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
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
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
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
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
structural investigation of a cell-wall galactomannan from neurospora crassa and n. sitophila. 19968901273
eth-1, the neurospora crassa locus encoding s-adenosylmethionine synthetase: molecular cloning, sequence analysis and in vivo overexpression.intense biochemical and genetic research on the eth-1r mutant of neurospora crassa suggested that this locus might encode s-adenosylmethionine synthetase (s-adomet synthetase). we have used protoplast transformation and phenotypic rescue of a thermosensitive phenotype associated with the eth-1r mutation to clone the locus. nucleotide sequence analysis demonstrated that it encodes s-adomet synthetase. homology analyses of prokaryotic, fungal and higher eukaryotic s-adomet synthetase polypeptide s ...19968849888
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
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
repeat-induced gene silencing: common mechanisms in plants and fungi.one of the most surprising observations made in plant science in recent years is the inactivation of transgenes triggered by interactions between dna repeats. in plants, we can differentiate between transcriptional silencing, most likely reflecting a regulation at the dna level, and post-transcriptional silencing that affects steady state rna levels. in the filamentous fungi ascobolus immersus and neurospora crassa, we find two premeiotic silencing processes that are also based on the interactio ...19968868065
activator-independent gene expression in neurospora crassa.a transgenic position effect that causes activator-independent gene expression has been described previously for three neurospora crassa phosphate-repressible genes. we report analogous findings for two additional positively regulated genes, qa-2+ and ars-1+, indicating that such position effects are not limited to genes involved in phosphorus metabolism. in addition, we have characterized a number of mutants that display activator-independent gene expression. each of these mutants contains a ch ...19968852841
properties of a polynucleotide synthesized by strain 74a of neurospora crassa.a polynucleotide (or a fragment of rna) was purified to apparent homogeneity by hplc from mycelium of the wild strain 74a of the mould neurospora crassa, after growth on sucrose and in the presence of saturating amounts of inorganic phosphate (pi) for 72 hr at 30 degrees. the m(r) was ca 20,000 as determined by hplc at ph 6.8. polynucleotide synthesis ranged from 4.0 to 6.5 micrograms polynucleotide per mg dry mycelium in mycelium of the wild strain 74a and the various phosphorus regulatory and ...19968821430
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
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
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
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
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
short communication: purification and properties of acid phosphatase (ec 3.1.3.2) secreted by strain 74a of the mould neurospora crassa.both pi-repressible acid phosphatases, iib (mycelial) and iic (extracellular), synthesized by neurospora crassa and purified to apparent homogeneity by 7.5% page, are monomers, are inhibited by 2 mm zncl2 and are non-specifically stimulated by salts. however, the iic form is activated by p-nitrophenylphosphate (in a negative co-operativity effect with a k 0.5 of 2.5 mm) whereas form iib shows michaelis kinetics, with a k m of 0.5 mm. thus, since both enzymatic forms may be expressed by the same ...199624415104
crystallization and preliminary crystallographic analysis of 3-carboxy-cis,cis-muconate lactonizing enzyme from neurospora crassa.crystals of 3-carboxy-cis,cis-muconate lactonizing enzyme (cmle; e.c. 5.5.1.5) from neurospora crassa that diffract to high resolution have been obtained. the crystals belong to the orthorhombic space group p2(1)2(1)2(1) with unit-cell dimensions a = 92.1, b = 159.7, c = 236.6 a (at 103 k) and diffract at most to 2 a resolution. the asymmetric unit of the crystals appears to contain two tetrameric cmle molecules making up a total of 328 kda per asymmetric unit. both cross-linking with glutaralde ...199615299752
repeat-induced point mutations of hsp80 gene of neurospora crassa: methylation of duplicated dna sequences in the vegetative state.the process of repeat-induced point mutations (rip) was used to disrupt the gene encoding the 80-kda heat-inducible protein of neurospora crassa. germinated conidia of the wild-type recipient strain were electrotransformed with a plasmid containing a 7-kb fragment harbouring the complete hsp80 gene sequence. some of the transformants with a duplication of hsp80 gene sequence showed extensive methylation of these sequences even in vegetatively growing cells. the presence of an extra gene copy in ...19969035689
disruption of the nad(+)-specific glutamate dehydrogenase gene of neurospora crassa by means of the rip (repeat-induced point mutations) process.the structural gene for the catabolite-repressed, substrate-induced nad(+)-specific glutamate dehydrogenase (gdh-1) of neurospora crassa was disrupted using the process of repeat-induced point mutation (rip). plasmids containing incomplete copies of the gene, along with selectable markers, were introduced into germinated conidia by electroporation. the sexual progeny of a transformant containing an ectopically integrated copy of a plasmid, harbouring the 5' flanking region and a part of the codi ...19969035688
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
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
genetic and enzymatic analysis of a glycerol kinase deficient mutant in neurospora crassa.genetic analysis showed that the glycerol non-utilizing isolate gly-u(234) of neurospora crassa is derived by mutation in a nuclear gene situated in the right arm of linkage group i, about 2.2 cross-over units distal to ad-9 and 11 units proximal to nit-1. enzymatic testings using a radiochemical method indicate that the mutant is deficient for the enzyme glycerol kinase. the radiochemical testings further indicate that the mutation has inactivated an inducible glycerol kinase, while a low resid ...1996177854
transport of cytoplasmically synthesized proteins into the mitochondria in a cell free system from neurospora crassa.synthesis and transport of mitochondrial proteins were followed in a cell-free homogenate of neurospora crassa in which mitochondrial translation was inhibited. proteins synthesized on cytoplasmic ribosomes are transferred into the mitochondrial fraction. the relative amounts of proteins which are transferred in vitro are comparable to those transferred in whole cells. cycloheximide and puromycin inhibit the synthesis of mitochondrial proteins but not their transfer into mitochondria. the transf ...1996145943
enzymes of deoxythymidine triphosphate biosynthesis in neurospora crassa mitochondria.intact mitochondria of neurospora crassa incorporate deoxythymidine 5'-monophosphate (dtmp) into deoxyribonucleic acid but not the label from (methyl-3h) deoxythymidine. mitochondrial homogenates contain deoxythymidylate kinase (ec 2.7.4.9), deoxycytidylate aminohydrolase (dcmp deaminase) (ec 3.5.4.12), and thymidylate synthetase (ec 2.1.1b), but not thymidine kinase (ec 2.7.1.21) activity. dtmp kinase is loosely bound to the mitochondrial membrane and is solubilized by 0.4 m kcl in mitochondria ...1996163227
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
identification of the native nit2 major nitrogen regulatory protein in nuclear extracts of neurospora crassa.the nit-2 gene of neurospora crassa encodes the major nitrogen regulatory protein which acts in a positive fashion to activate the expression of many different structural genes during conditions of nitrogen limitation. an e. coli-expressed nit2/beta-gal fusion protein binds specifically to dna in vitro by recognizing gata core elements. nuclear extracts prepared from a wild-type n. crassa strain contain a protein factor which displays all of the properties expected for the native nit2 protein. t ...19968901135
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
covalent attachment of fad derivatives to a fusion protein consisting of 6-hydroxy-d-nicotine oxidase and a mitochondrial presequence. folding, enzyme activity, and import of the modified protein into yeast mitochondria.autoflavinylation of 6-hydroxy-d-nicotine oxidase (6-hdno) was successfully employed to modify the protein covalently with fad derivatives. the model compounds n6-(2-aminoethyl)-fad and n6-(6-carboxyhexyl)-fad were spontaneously bound to a fusion protein consisting of the mitochondrial targeting sequence of neurospora crassa f0-atpase subunit 9 (su9) attached to 6-hdno. when translated in the rabbit reticulocyte lysate, su9-6-hdno was in the trypsin-sensitive apoenzyme form; when translated in t ...19968810280
a transposon insertion in the arabidopsis ssr16 gene causes an embryo-defective lethal mutation.the ssr16 gene of arabidopsis has been identified as a gene encoding a ribosomal protein s16 homolog through analysis of a transposon insertion mutation. the insertion mutation is lethal, arresting embryonic development at approximately the transition from the globular to the heart stage of embryonic development. co-segregation of the mutant phenotype with the transposon-borne drug-resistance marker and loss of the inserted transposon concomitant with phenotypic reversion provided evidence that ...19968811862
the isolation of a dol-p-man synthase from ustilago maydis that functions in saccharomyces cerevisiae.genomic dnas from several fungi were screened for a homologous sequence to saccharomyces cerevisiae dpm1, an essential gene which encodes dolichyl phosphoryl mannose synthase. the fungi examined included aspergillus nidulans, neurospora crassa, schizophyllum commune and ustilago maydis. only u. maydis gave a significant signal after southern hybridization using dpm1 as a probe. the ustilago homolog was subsequently cloned and sequenced. the predicted protein of 294 amino acids has 60% identity t ...19968813763
genetic variability among isolates and sexual offspring of the plant pathogenic fungus calonectria morganii on the basis of random amplification of polymorphic dna (rapd) and restriction fragment length polymorphism (rflp)thirty-two strains of the phytopathogenic mold cylindrocladium scoparium (perfect state calonectria morganii) isolated from ericaceous hosts and two specimens from the atcc were examined by random amplification of polymorphic dna (rapd) and restriction fragment length polymorphism (rflp). five oligonucleotides were chosen as primers for differentiation of the isolates. rapd patterns of the atcc strains differed significantly from those of the field isolates. diversity among field isolates was lo ...19968824171
a tyrosyl-trna synthetase suppresses structural defects in the two major helical domains of the group i intron catalytic core.the neurospora crassa mitochondrial tyrosyl-trna synthetase, the cyt-18 protein, functions in splicing group i introns by promoting the formation of the catalytically active structure of the intron rna. the group i intron catalytic core is thought to consist of two extended helical domains, one formed by coaxial stacking of p5, p4, p6, and p6a (p4-p6 domain) and the other consisting of p8, p3, p7, and p9 (p3-p9 domain). to investigate how cyt-18 stabilizes the active rna structure, we used an es ...19968831782
isolation of laccase gene-specific sequences from white rot and brown rot fungi by pcr.degenerate primers corresponding to the consensus sequences of the copper-binding regions in the n-terminal domains of known basidiomycete laccases were used to isolate laccase gene-specific sequences from strains representing nine genera of wood rot fungi. all except three gave the expected pcr product of about 200 bp. computer searches of the databases identified the sequence of each of the pcr products analyzed as a laccase gene sequence, suggesting the specificity of the primers. pcr product ...19968837429
development of a metallothionein based heavy metal biosorbent.the potential utility of a recombinant e. coli expressing the neurospora crassa metallothionein gene (ncp) as a heavy metal biosorbent was investigated. it was shown that the ncp was capable of efficiently removing low levels of several metals (including cadmium, lead, and mercury) from solutions. the reusability of the ncp was demonstrated through 5 cycles of metal binding, stripping with dilute acid, and regeneration of the binding sites with out any adverse effect on the metal binding activit ...19968843348
in vivo calcineurin crystals formed using the baculovirus expression system.calcineurin is a heterodimeric phosphatase involved in the signal transduction of antigen-activated t cells. coexpression of its two subunits, the regulatory subunit from human and the catalytic subunit from neurospora crassa in cultured insect cells using the baculovirus expression system results in the formation of very large crystals in the cytoplasm. the crystals are formed initially in vesicles, but their subsequent growth appears to be uninhibited and continues without the need of an enclo ...19968859891
nuva, an aspergillus nidulans gene involved in dna repair and recombination, is a homologue of saccharomyces cerevisiae rad18 and neurospora crassa uvs-2.a 40 kb genomic clone and 2.3 kb ecori subclone that rescued the dna repair and recombination defects of the aspergillus nidulans nuva11 mutant were isolated and the subclone sequenced. the subclone hybridized to a cosmid in a chromosome-specific library confirming the assignment of nuva to linkage group iv and indicating its closeness to bimd. amplification by pcr clarified the relative positions of nuva and bimd. a region identified within the subclone, encoding a c3hc4 zinc finger motif, was ...19968868425
m-glycogenin, the protein moiety of neurospora crassa proteoglycogen, is an auto- and transglucosylating enzyme.neurospora crassa proteoglycogen was purified and its protein moiety, m-glycogenin, was released by amylolytic treatment. the released protein was capable of autoglucosylation from udp-glucose forming glucosyl-alpha 1,4-glucosyl linkage. the kinetics of autoglucosylation suggested an intramolecular mechanism of reaction. m-glycogenin was also able to glucosylate dodecyl-beta-maltoside and autoglucosylate, simultaneously and independently. both auto- and transglucosylation reactions were dependen ...19968886029
characterization of rco-1 of neurospora crassa, a pleiotropic gene affecting growth and development that encodes a homolog of tup1 of saccharomyces cerevisiae.the filamentous fungus neurospora crassa undergoes a well-defined developmental program, conidiation, that culminates in the production of numerous asexual spores, conidia. several cloned genes, including con-10, are expressed during conidiation but not during mycelial growth. using a previously described selection strategy, we isolated mutants that express con-10 during mycelial growth. selection was based on expression of an integrated dna fragment containing the con-10 promoter-regulatory reg ...19968887652
evolution of the frequency (frq) clock locus in ascomycete fungi.the frequency (frq) locus of neurospora crassa plays a key role in the organization of circadian rhythms. similar timing systems have been found in nearly all eukaryotes as well as some prokaryotes; thus, frq may be an excellent gene with which to conduct evolutionary studies. to investigate, we used the cloned frq locus from ascomycete fungi representing two classical taxonomic classes and three orders to examine two open questions in ascomycete evolution. class pyrenomycetidae is represented b ...19968896376
functional in vivo studies of the neurospora crassa cys-14 gene upstream region: importance of cys3-binding sites for regulated expression.sulphate transport in neurospora crassa is achieved by two distinct sulphate permeases, i and ii, encoded by the cys-13 and cys-14 genes, respectively. the synthesis of both sulphate permeases is subject to sulphur repression and requires the global positive-acting regulatory protein cys3, cys3, a bzip dna binding protein, regulates cys-14 expression at the transcriptional level and binds in vitro specifically to three dna-recognition sites, a, b, and c, in the cys-14 upstream region. in vivo fu ...19968899713
asm-1+, a neurospora crassa gene related to transcriptional regulators of fungal development.this report describes the identification, cloning, and molecular analysis of asm-1+ (ascospore maturation 1), the neurospora crassa homologue of the aspergillus nidulans stua (stunted a) gene. the asm-1+ gene is constitutively transcribed and encodes an abundant, nucleus-localized 68.5-kd protein. the protein product of asm-1+ (asm-1), contains a potential dna-binding motif present in related proteins from a. nidulans (stua), candida albicans (efgtf-1), and saccharomyces cerevisiae (phd1 and sok ...19968913744
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