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in vivo overproduction of the pentafunctional arom polypeptide in aspergillus nidulans affects metabolic flux in the quinate pathway.the shikimate pathway and the quinic acid utilisation (qut) pathway of aspergillus nidulans and other fungi share the two common metabolic intermediates, 3-dehydroquinic acid (dhq) and dehydroshikimic acid (dhs), which are interconverted by two isoenzymes, catabolic 3-dehydroquinase, (cdhqase) and biosynthetic dehydroquinase, (bdhqase). bdhqase is one of five consecutive enzymatic activities associated with the pentafunctional arom protein encoded by the complex arom locus, whereas cdhqase is en ...19911648168
a cdna encoding rabbit muscle protein phosphatase 1 alpha complements the aspergillus cell cycle mutation, bimg11.the bimg11 allele causes a conditional growth defect in the fungus aspergillus nidulans preventing both progression through mitosis and normal polar growth. previously, we have shown that the bimg11 mutation increases the phosphorylation of nuclear proteins and that the gene encodes a protein similar to mammalian type 1 protein phosphatase. assay of protein phosphatase activity in protein extracts of aspergillus demonstrates directly that type 1 phosphatase activity is greatly reduced in the mut ...19911655766
diverse proteins homologous to inositol monophosphatase.bovine inositol monophosphatase (imp) and several homologous proteins were found to share two sequence motifs with bovine inositol polyphosphate 1-phosphatase (ipp). these motifs may correspond to binding sites within imp and ipp for inositol phosphates or for lithium, since both substances are bound by these proteins. this suggests that the proteins homologous to imp, which have diverse biological roles but whose function is not clear, may act by enhancing the synthesis or degradation of phosph ...19911660408
two alpha-tubulin genes of aspergillus nidulans encode divergent proteins.we have isolated and analyzed the tuba and tubb alpha-tubulin genes of aspergillus nidulans. the nucleotide sequences of these genes predict polypeptides of 447 amino acids for tuba and 450 for tubb. the predicted amino acids sequences exhibit 28% divergence between the two polypeptides. this is the second known case of such high divergence between alpha-tubulins within the same species. the tubb gene is unique in that it codes for an extra glycine residue between what are usually the second and ...19911672037
is there methylmalonyl coa mutase in aspergillus nidulans?in most animal species and many prokaryotes, methylmalonyl coa mutase catalyzes isomerization between methylmalonyl coa and succinyl coa using adenosylcobalamin as a cofactor. we describe the absence of this enzyme in aspergillus nidulans based on the absence of enzyme activity in vitro and the failure to metabolize methylmalonate or grow in media containing this organic acid as the sole carbon source. these data contrast previous assumptions that propionate may be metabolized through propionyl ...19911676260
isolation and expression of the acetate-inducible isocitrate lyase gene (acu-3) from neurospora crassa: evidence for a second constitutive isozyme.heterologous hybridisation of the aspergillus nidulans structural gene for isocitrate lyase (acud) to a lambda genomic library of neurospora crassa identified a recombinant phage containing the hybridising sequence on an internal 9 kb ecori fragment. a restriction fragment length polymorphism (rflp) enabled the fragment to be assigned to linkage group v (lg v), the location of the acetate-inducible isocitrate lyase, acu-3 of neurospora. functional ectopic complementation by co-transformation of ...19911681413
sequence and expression of gln3, a positive nitrogen regulatory gene of saccharomyces cerevisiae encoding a protein with a putative zinc finger dna-binding domain.the gln3 gene of saccharomyces cerevisiae is required for the activation of transcription of a number of genes in response to the replacement of glutamine by glutamate as source of nitrogen. we cloned the gln3 gene and constructed null alleles by gene disruption. gln3 is not essential for growth, but increased copies of gln3 lead to a drastic decrease in growth rate. the complete nucleotide sequence of the gln3 gene was determined, revealing one open reading frame encoding a polypeptide of 730 a ...19911682800
cloning by differential screening of a xenopus cdna that encodes a kinesin-related protein.by differential screening of a xenopus egg cdna library, we selected nine clones (eg1 to eg9) corresponding to mrnas which are deadenylated and released from polysomes soon after fertilization. the sequence of one of these clones (eg5) revealed that the corresponding protein has the characteristic features of a kinesin-related protein. more specifically, eg5 was found to be nearly 30% identical to a kinesin-related protein encoded by bimc, a gene involved in nuclear division in aspergillus nidul ...19911710028
effect of citrate on radial growth and conidiation of the mould aspergillus nidulans.a mutation of the ctsa locus of aspergillus nidulans affects both the radial growth and conidiation of the mould when grown in the presence of citrate. the ctsa locus was allocated to linkage group iv but it recombines freely with inob2 and pyroa4 (which are also in linkage group iv). it is recessive in heterozygous diploids. a possible role for this gene in maintaining membrane integrity is discussed.199124425277
characterization of aspergillus nidulans mutants deficient in cell wall chitin or glucan.by screening for the osmotically remediable phenotype, mutations in two genes (orla and orlb) affecting the cell wall chitin content of aspergillus nidulans were identified. strains carrying temperature-sensitive alleles of these genes produce conidia which swell excessively and lyse when germinated at restrictive temperatures. growth under these conditions is remedied by osmotic stabilizers and by n-acetylglucosamine (glcnac). remediation by glcnac suggests that the mutations affect early steps ...19921729232
molecular characterization of the lam locus and sequences involved in regulation by the amdr protein of aspergillus nidulans.the lam locus of aspergillus nidulans consists of two divergently transcribed genes, lama and lamb, involved in the utilization of lactams such as 2-pyrrolidinone. both genes are under the control of the positive regulatory gene amdr and are subject to carbon and nitrogen metabolite repression. the lamb gene and the region between the two genes have been sequenced, and the start points of transcription have been determined. within the lam locus are two sequences with homology to elements, requir ...19921729609
an intragenic map of the brla locus of aspergillus nidulans.we have constructed an intragenic map for the aspergillus nidulans brla gene, mutants in which are distinguishable by visual criteria only. most of the leaky phenotype mutants map near the right (3') end. the gene shows distinct recombinational polarity consistent with recombination initiation at the promoter (centromere-proximal) end of the gene. brla12 and brla20 mutants gave abnormal dna restriction patterns consistent with the iii; viii and vi; viii translocations, respectively, determined b ...19921736092
tests for recombinagens in fungi.three types of mitotic recombination can be studied in aspergillus nidulans and saccharomyces cerevisiae: (1) the classical type of reciprocal mitotic crossing-over which can be detected when it occurs between non-sister chromatids at the four-strand stage followed by co-segregation of a crossing-over and a non-crossing-over chromatid in the subsequent mitotic division. consequently, mitotic crossing-over reflects cellular responses to primary genetic damage in the g2 phase of the cell cycle. (2 ...19921279386
immunogenic potential of aspergillus nidulans subcellular fractions and their polypeptide components.cell-free extracts of the ascomycetous fungus aspergillus nidulans were separated into three subcellular fractions: cell walls, total membranes and cytosol, and two different immunization protocols were used to raise antibodies against them in 12 new zealand rabbits. the immune response was followed over time by dot and western blot analyses to determine the immunogenic potential of each individual fraction and their polypeptide components. the igg fractions, purified from pools of the best sera ...19921291875
structure of the oxidized long-chain flavodoxin from anabaena 7120 at 2 a resolution.the structure of the long-chain flavodoxin from the photosynthetic cyanobacterium anabaena 7120 has been determined at 2 a resolution by the molecular replacement method using the atomic coordinates of the long-chain flavodoxin from anacystis nidulans. the structure of a third long-chain flavodoxin from chondrus crispus has recently been reported. crystals of oxidized a. 7120 flavodoxin belong to the monoclinic space group p2(1) with a = 48.0, b = 32.0, c = 51.6 a, and beta = 92 degrees, and one ...19921303762
roles of the orla, tse, and bimg genes of aspergillus nidulans in chitin synthesis.strains of aspergillus nidulans carrying the orla1 or tse6 allele are deficient in cell wall chitin and undergo lysis at restrictive temperatures. the strains are remediable by osmotic stabilizers or by the presence of n-acetylglucosamine (glcnac) in the medium. the remediation by glcnac suggests that the lesion(s) in chitin synthesis resides in the amino sugar biosynthetic pathway prior to the synthesis of n-acetylglucosamine-6-phosphate. orla1 strains grown at permissive temperature exhibit an ...19921309526
differential flux through the quinate and shikimate pathways. implications for the channelling hypothesis.the qutc gene encoding dehydroshikimate dehydratase has been constitutively overexpressed in aspergillus nidulans from a range of 1-30-fold over the normal wild-type level. this overexpression leads to impaired growth in minimal medium which can be alleviated by the addition of aromatic amino acids to the medium. overexpression of the qutc gene in mutant strains lacking protocatechuic acid (pca) oxygenase leads to the build up of pca in the medium, which can be measured by a simple assay. measur ...19921318019
analysis of promoter activity by transformation of acremonium chrysogenum.promoter activity was examined in the beta-lactam-producing fungus, acremonium chrysogenum, by assessment of the properties of transformant isolates. transformation was achieved using plasmid constructs specifying hygromycin b resistance (hyr) linked to the promoter elements of gpda (the glucose-6-phosphate dehydrogenase-encoding gene of aspergillus nidulans), and pcbc [the gene encoding the isopenicillin n synthetase (ipns) enzyme of a. chrysogenum]. transformation frequency, hyr levels, and hy ...19921318244
overproduction in escherichia coli of the dehydroquinate synthase domain of the aspergillus nidulans pentafunctional arom protein.the pentafunctional arom protein of aspergillus nidulans is encoded by the complex aroma locus and catalyses steps 2-6 in the synthesis of chorismate, the common precursor for the aromatic amino acids and p-aminobenzoic acid. dna sequences encoding the 3-dehydroquinate synthase (dhq synthase) and 3-dehydroquinase domains of the arom protein have been amplified with the inclusion of a translational stop codon at the c-terminus by pcr technology. these amplified fragments of dna have been subclone ...19921320381
purification and characterization of a mammalian endo-exonuclease.an endo-exonuclease has been purified from cultured monkey (cv-1) cells. the enzyme which was purified to near homogeneity to be a 65 kda monomeric protein. the single-strand dnase activity is endonucleolytic and nonprocessive, whereas the double-strand dnase activity is exonucleolytic and processive. the enzyme was also found to have rnase activity using poly-ra as substrate. the ph optimum for ss-dnase is 8 and for ds-dnase it is 7.5. both dnase activities require a divalent metal ion (mg2+, m ...19921324480
mitochondrial dna sequence analysis of the cytochrome oxidase subunit i and ii genes, the atpase9 gene, the nadh dehydrogenase nd4l and nd5 gene complex, and the glutaminyl, methionyl and arginyl trna genes from trichophyton rubrum.in this paper, we present the nucleotide sequence of a 5248 bp-long region of the mitochondrial (mt) genome of the dermatophyte trichophyton rubrum. this region which represents about 1/4 of the total mt genome of this species reveals a compact organization of genes including: the glutaminyl trna, the methionyl trna, the cytochrome oxidase subunit i gene, the arginyl trna, the mitochondrial version of the atpase subunit 9 gene, the cytochrome oxidase subunit ii gene and a part of the nadh dehydr ...19921326416
transformation of gibberella fujikuroi: effect of the aspergillus nidulans ama1 sequence on frequency and integration.a stable and reproducible transformation selection system for gibberella fujikuroi protoplasts based on the aspergillus nidulans arg b gene, encoding ornithine transcarbamylase, has been developed. inclusion into the vector of the a. nidulans dna fragment (ama1), which permits plasmid autonomous replication in a. nidulans, a. niger and a. oryzae, appeared to permit autonomous replication of g. fujikuroi although the transformation frequency was increased by only two-fold. transformation was also ...19921327547
inducible overproduction of the aspergillus nidulans pentafunctional arom protein and the type-i and -ii 3-dehydroquinases from salmonella typhi and mycobacterium tuberculosis.the aroq gene of mycobacterium tuberculosis, encoding a type-ii 3-dehydroquinase, and the arod gene of salmonella typhi, encoding a type-i 3-dehydroquinase, have been highly overexpressed in escherichia coli using the powerful trc promoter contained within the expression vector pkk233-2. the m. tuberculosis type-ii 3-dehydroquinase has been purified in bulk from overproducing strains of e. coli to greater than 95% homogeneity. the protein is extremely heat-stable, is active as a homododecamer an ...19921329726
cell division in aspergillus.amenable to sophisticated genetic and molecular analysis, the simple filamentous fungus aspergillus nidulans has provided some novel insights into the mechanisms and regulation of cell division. mutational analysis has identified over fifty genes necessary for nuclear division, nuclear movement and cytokinesis. molecular and cellular analysis of these mutants has led to the discovery of novel components of the cytoskeleton as well as to clarifying the role of established cytoskeletal proteins. m ...19921336015
structure of the cochliobolus heterostrophus glyceraldehyde-3-phosphate dehydrogenase gene.a single gene (gpd1) encoding glyceraldehyde-3-phosphate dehydrogenase (gpd) was found in cochliobolus heterostrophus. homology with other fungal gpd-encoding genes was substantial at both the nucleotide and amino-acid levels. positions of four introns found in gpd1 were conserved in the corresponding aspergillus nidulans gpda gene (which is known to have three additional introns absent in gpd1). the size (approximately 1300 nucleotides) of the single gpd1 transcript was consistent with the leng ...19921339326
a second gene (quth) within the aspergillus nidulans-quinic-acid utilisation gene cluster encodes a protein with a putative zinc-cluster motif.a sequence of 3299 nt, contiguous with the previously sequenced quinate permease-encoding (qutd) gene and encompassing the dehydroshikimate dehydratase-encoding (qutc) gene, has been determined. northern-blot analysis detected (i) a quinate-inducible mrna of the expected size for the qutc gene, and (ii) a quinate-inducible mrna of 1.45 kb divergently transcribed away from qutc towards qutd. computer-aided sequence analysis identified an orf of 1047 nt corresponding to the qutc gene encoding dehy ...19921339361
calmodulin and cell cycle control.previous studies have indicated a role for the calcium receptor calmodulin in the control of eukaryotic cell proliferation. using a molecular genetic approach in the filamentous fungus aspergillus nidulans we have shown that cam is required for cell cycle progression at multiple points in the cell cycle. construction of an a nidulans strain conditional for calmodulin expression reveals that this protein is required during g1/s and for the initiation of mitosis. a lack of calmodulin results in ce ...19921343599
genotoxicity of fungi evaluated by sos microplate assay.by an introduction of sodium dodecylsulfate for cell lysis and immunomicroplate for mass assay, the modified sos microplate assay method was established and applied for the evaluation of genotoxicity of mycotoxins and fungal cultures. among 20 mycotoxins, the carcinogenic dihydrobisfuranoids such as aflatoxin b1, sterigmatocystin, and versicolorin a were positive in the presence of the activation system. while, the carcinogenic anthraquinones and lactones such as luteoskyrin, rugulosin, ochratox ...19921344897
isolation and characterization of the acetyl-coa synthetase from penicillium chrysogenum. involvement of this enzyme in the biosynthesis of penicillins.acetyl-coa synthetase (acs) of penicillium chrysogenum was purified to homogeneity (745-fold) from fungal cultures grown in a chemically defined medium containing acetate as the main carbon source. the enzyme showed maximal rate of catalysis when incubated in 50 mm hcl-tris buffer, ph 8.0, at 37 degrees c. under these conditions, acs showed hyperbolic behavior against acetate, coa, and atp; the km values calculated for these substrates were 6.8, 0.18, and 17 mm, respectively. acs recognized as s ...19921347531
nucleotide sequence of a gene from phanerochaete chrysosporium that shows homology to the faca gene of aspergillus nidulans.heterologous hybridisation was used to isolate a genomic dna sequence from phanerochaete chrysosporium using the faca (acetyl coa synthetase) gene from aspergillus nidulans as a probe. the cloned sequence hybridises to a 2.2 kb transcript in poly(a)+ rna prepared from mycelium grown on acetate as the sole carbon source. comparison of the dna sequence obtained with those of the a. nidulans faca and n. crassa acu5 genes reveals an orf that appears to be interrupted by five typical fungal introns. ...19921352996
the acu-1 gene of coprinus cinereus is a regulatory gene required for induction of acetate utilisation enzymes.we have isolated a gene from coprinus cinereus which cross-hybridises to the faca and acu-5 genes of aspergillus nidulans and neurospora crassa, respectively. these genes encode acetyl-coa synthetase, an enzyme which is inducible by acetate and required for growth on acetate as sole carbon source. we have designated the c. cinereus gene acs-1 and have used transformation to demonstrate its functional homology to the ascomycete genes by complementation of an n. crassa acu-5 mutation. the acs-1 ge ...19921354839
mitochondrial dna of schizophyllum commune: restriction map, genetic map, and mode of inheritance.mitochondrial dna (mtdna) found in the basidiomycete schizophyllum commune (strain 4-40) is a circular molecule 49.75 kbp in length. a physical map containing 61 restriction sites revealed no repeat structures. cloned genes from neurospora crassa, aspergillus nidulans, and saccharomyces cerevisiae were used in southern hybridizations to locate nine mitochondrial genes, including a possible pseudogene of atpase 9, on the restriction map. a probe from a functional atpase 9 gene identified homologo ...19921358467
cloning and disruption of a gene required for growth on acetate but not on ethanol: the acetyl-coenzyme a synthetase gene of saccharomyces cerevisiae.a dna fragment of saccharomyces cerevisiae with high homology to the acetyl-coenzyme a (acetyl-coa) synthetase genes of aspergillus nidulans and neurospora crassa has been cloned, sequenced and mapped to chromosome i. it contains an open reading frame of 2139 nucleotides, encoding a predicted gene product of 79.2 kda. in contrast to its ascomycete homologs, there are no introns in the coding sequence. the first atg codon of the open reading frame is in an unusual context for a translational star ...19921363452
clusters of genes for the biosynthesis of antibiotics: regulatory genes and overproduction of pharmaceuticals.in the last decade numerous genes involved in the biosynthesis of antibiotics, pigments, herbicides and other secondary metabolites have been cloned. the genes involved in the biosynthesis of penicillin, cephalosporin and cephamycins are organized in clusters as occurs also with the biosynthetic genes of other antibiotics and secondary metabolites (see review by martín and liras [65]). we have cloned genes involved in the biosynthesis of beta-lactam antibiotics from five different beta-lactam pr ...19921368054
mitotic gold in a mold: aspergillus genetics and the biology of mitosis.the analysis of fungal mutants has had an extraordinary impact on our understanding of the biochemistry and regulation of mitosis. in this article we review the contribution of work on the filamentous fungus aspergillus nidulans to the molecular genetics of mitosis.19921369734
l-lysine repression of penicillin biosynthesis and the expression of penicillin biosynthesis genes acva and ipna in aspergillus nidulans.the addition of 0.1 m l-lysine to the fermentation medium reduced the production of penicillin by about 50% in aspergillus nidulans. to analyse this effect at the molecular level, the expression of the penicillin biosynthesis genes acva and ipna, encoding delta-(l-alpha-aminoadipyl)-l-cysteinyl-d-valine synthetase and isopenicillin n synthetase, was studied by using translational fusions with different reporter genes (strain axb4a, acva-uida, ipna-lacz fusions; axb4b, acva-lacz, ipna-uida fusion ...19921369977
the induction of mitotic chromosome malsegregation in aspergillus nidulans. quantitative structure activity relationship (osar) analysis with chlorinated aliphatic hydrocarbons.the biological activity of 24 chlorinated aliphatic hydrocarbons has been studied in the mold aspergillus nidulans. the ability to induce chromosome malsegregation, lethality and mitotic growth arrest has been experimentally determined for each chemical. these data, together with those of 11 related compounds previously investigated, generated a data base which was used for quantitative structure-activity relationship (qsar) analysis. to this aim, both physico-chemical descriptors and electronic ...19921373821
cloning and properties of a cyanide hydratase gene from the phytopathogenic fungus gloeocercospora sorghi.the cht gene encoding cyanide hydratase (cht, ec 4.2.1.66), which detoxifies hcn and is thought to be important in fungal infection of cyanogenic plants, has been cloned from the phytopathogenic fungus gloeocercospora sorghi. the gene was isolated by screening an expression library of g. sorghi using a cht-specific antibody and using one of the positive cdna clones as a probe in southern hybridization to identify a 3.1 kb psti genomic fragment. this psti fragment expressed cht activity when tran ...19921382413
a mammalian dual specificity protein kinase, nek1, is related to the nima cell cycle regulator and highly expressed in meiotic germ cells.screening of mouse cdna expression libraries with antibodies to phosphotyrosine resulted in repeated isolation of cdnas that encode a novel mammalian protein kinase of 774 amino acids, termed nek1. nek1 contains an n-terminal protein kinase domain which is most similar (42% identity) to the catalytic domain of nima, a protein kinase which controls initiation of mitosis in aspergillus nidulans. in addition, both nek1 and nima have a long, basic c-terminal extension, and are therefore similar in o ...19921382974
reversible inactivation of a foreign gene, hph, during the asexual cycle in neurospora crassa transformants.a plasmid construct carrying the hygromycin phosphotransferase (hph) gene fused to the expression elements of the trpc gene of aspergillus nidulans was used to obtain hygromycin b (hyg)-resistant transformants of neurospora crassa. the plasmid does not have any homology with the n. crassa genome. here we demonstrate that most of the transformants arise from integration of the transforming dna into only one of the nuclei present in the protoplasts. furthermore, in most of the transformants the in ...19921383683
transformation of cochliobolus lunatus with put 720 changes the steroid hydroxylating ability of the fungus.the filamentous fungus cochliobolus lunatus, a known 11 beta-hydroxylator of steroids, was transformed to bleomycin resistance using the heterologous plasmid put 720. this plasmid contains the sh ble gene expressed under the control of the aspergillus nidulans gpd and trpc expression signals. the bleomycin-resistant colonies appeared with a frequency of six per microgram of dna. all colonies were real transformants and no "abortive" growth was observed. in all transformants tested the plasmid mo ...19921384995
an nadp(+)-dependent glycerol dehydrogenase in aspergillus nidulans is inducible by d-galacturonate.in aspergillus nidulans there is an nadp(+)-dependent glycerol dehydrogenase that is specifically induced on transfer to d-galacturonate medium. in contrast to the previously characterised constitutive nadp(+)-dependent glycerol dehydrogenase it has a much broader substrate specificity, having activity as an ethanol dehydrogenase, and is subject to carbon-catabolite repression. in addition to the two nadp(+)-dependent glycerol dehydrogenases, alcohol dehydrogenase i and ii are also present on tr ...19921394511
an upstream activating sequence from the aspergillus nidulans gpda gene.introduction of a previously identified promoter element of the aspergillus nidulans gpda gene (encoding glyceraldehyde-3-phosphate dehydrogenase), the so-called gpd box, into the upstream region of the highly regulated a. nidulans amds gene (encoding acetamidase), significantly increased (up to 30-fold) the expression of the lacz reporter gene fused to these expression signals. this increase was dependent on the orientation of the gpd box and on the site of introduction into the amds upstream r ...19921398125
resolution of chromosomes iii and vi of aspergillus nidulans by pulsed-field gel electrophoresis shows that the penicillin biosynthetic pathway genes pcbab, pcbc, and pende are clustered on chromosome vi (3.0 megabases).an improved electrophoretic molecular karyotype of aspergillus nidulans atcc 28901 has been obtained by contour-clamped electric field gel electrophoresis, which separates seven chromosomal bands and allows resolution of chromosomes iii and vi. the three genes of the penicillin biosynthetic pathway, pcbab, pcbc, and pende, encoding alpha-aminoadipyl-cysteinyl-valine synthetase, isopenicillin n synthase, and isopenicillin n acyltransferase, respectively, are clustered together on a chromosome of ...19921400258
specific binding sites for the activator protein, alcr, in the alca promoter of the ethanol regulon of aspergillus nidulans.alcr is the specific activator of the aspergillus nidulans ethanol-utilization pathway, mediating the induction of its own transcription and that of the structural genes alca and alda, encoding respectively, alcohol dehydrogenase i and aldehyde dehydrogenase. alcr is a dna binding protein in which 6 cysteines are coordinated in a zinc binuclear cluster. this domain was fused to glutathione-s-transferase (gst) and isolated as a gst-alcr(7-58*) fusion protein from escherichia coli. mobility shift ...19921400424
development of a transformation system for the thermophilic fungus talaromyces sp. cl240 based on the use of phleomycin resistance as a dominant selectable marker.a transformation system for the thermophilic cellulolytic fungus talaromyces sp. cl240 has been developed, using the phleomycin resistance gene from streptoalloteichus hindustanus (sh ble) as a dominant selectable marker. the plasmids (pan8-1 and put720) carrying the sh ble gene under the control of the aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter, allowed selection of phleomycin-resistant transformants. a new promoter sequence cloned from chromosomal dna of trich ...19921406595
cmap: contig mapping and analysis package, a relational database for chromosome reconstruction.in the contig mapping and analysis package, cmap, we provide a foundation for reverse genetics by organizing information about dna fragments obtained from an organism's genome into a physical map. the user can store information about a particular segment of dna. this information can be both descriptive, such as any genes contained in a particular dna fragment, or experimental, such as hybridization profiles or restriction digest patterns for comparison with other fragments. the package can then ...19921422880
amino acid alterations in the bena (beta-tubulin) gene of aspergillus nidulans that confer benomyl resistance.we report the cloning and sequencing of 18 mutant alleles of the bena, beta-tubulin gene of aspergillus nidulans that confer resistance to the benzimidazole antifungal, antimicrotubule compounds benomyl, carbendazim, nocodazole, and thiabendazole. in 12 cases, amino acid 6 was changed from histidine to tyrosine or leucine. in four cases, amino acid 198 was changed from glutamic acid to aspartic acid, glutamine, or lysine. in two cases, amino acid 200 was altered from phenylalanine to tyrosine. t ...19921423663
expression of a bacterial aspartase gene in aspergillus nidulans: an efficient system for selecting multicopy transformants.the escherichia coli aspartase gene aspa has been expressed in the fungus aspergillus nidulans using the powerful constitutive gpda promoter and trpc terminator, both from a. nidulans. multiple, but not single, copies of aspa overcome nutritional deficiencies resulting from the loss of catabolic nad-linked glutamate dehydrogenase. they also circumvent certain nutritional deficiencies resulting from loss of the positive-acting regulatory gene product mediating nitrogen metabolite repression. both ...19921423725
transformation of acremonium coenophialum, a protective fungal symbiont of the grass festuca arundinacea.acremonium coenophialum is a mutualistic mycosymbiont and natural agent of biological protection of the widely distributed grass festuca arundinacea (tall fescue). an electroporative transformation system was developed for a. coenophialum. segments of dna 5' to the beta-tubulin gene (tub2) of the closely related ascomycete epichloë typhina, fused to the escherichia coli hph gene encoding hygromycin b phosphotransferase, conferred hygromycin resistance when introduced into a. coenophialum by elec ...19921423727
purification and characterization of two extracellular beta-glucosidases from aspergillus nidulans.two beta-glucosidases, p-i and p-ii, were purified from the culture filtrate of aspergillus nidulans. the m(r) values of p-i and p-ii were about 125,000 and 50,000, respectively. the isoelectric point, optimal ph and temperature, michaelis constants for several substrates and inhibition constants for glucose and glucono-delta-lactone of each enzyme were determined. we conclude that the high affinity toward cellobiose and low inhibition by glucose of these enzymes may offer significant advantages ...19921426998
the use of simulated annealing in chromosome reconstruction experiments based on binary scoring.we present a method of combinatorial optimization, simulated annealing, to order clones in a library with respect to their position along a chromosome. this ordering method relies on scoring each clone for the presence or absence of specific target sequences, thereby assigning a digital signature to each clone. specifically, we consider the hybridization of oligonucleotide probes to a clone to constitute the signature. in that the degree of clonal overlap is reflected in the similarity of their ...19921427046
selective expression of a major allergen and cytotoxin, asp f i, in aspergillus fumigatus. implications for the immunopathogenesis of aspergillus-related diseases.asp f i is a major 18-kda aspergillus fumigatus allergen and a member of the mitogillin family of cytotoxins. the nucleotide sequence of the asp f i gene was determined by sequencing polymerase chain reaction products amplified from a. fumigatus spore dna. the entire 678-bp dna includes an 81-bp leader sequence, preceding the n-terminal alanine codon, a 52-bp intron, and a 444-bp open reading frame, encoding a 149-amino acid protein (m(r) 16,899), which is 99% homologous to mitogillin from asper ...19921431110
analysis of the site of action of the amdr product for regulation of the amds gene of aspergillus nidulans.the amdr gene of aspergillus nidulans regulates a number of structural genes in response to omega amino acid inducers. the site of action of the amdr product on expression of the amds gene was investigated by studying the effects of changes in the 5' region of amds, generated in vitro, on the induction, and on responses of an amds-lacz fusion gene to an amdrc allele. a sequence was identified that is sufficient for amdr regulation and that shows identity with sequences involved in amdr regulatio ...19921435733
prediction of transmembrane topology of f0 proteins from escherichia coli f1f0 atp synthase using variational and hydrophobic moment analyses.the a subunit, a membrane protein from the e. coli f1f0 atp synthase has been examined by fourier analysis of hydrophobicity and of amino-acid residue variation. the amino-acid sequences of homologous subunits from vibrio alginolyticus, saccharomyces cerevisiae, neurospora crassa, aspergillus nidulans, schizosaccharomyces pombe and candida parapsilosis were used in the variability analysis. by fourier analysis of sequence variation, two transmembrane helices are predicted to have one face in con ...19921445940
structural and functional analysis of the amdr regulatory gene of aspergillus oryzae.we have isolated the aspergillus oryzae homologue of the amdr regulatory gene of aspergillus nidulans by cross hybridization. sequence analysis and functional studies have shown that the amdr genes are highly conserved and functionally interchangeable between the two species. the homology between the two genes extends throughout most of the coding sequences, including sequences encoding the dna-binding domain and putative activation domains. two regions of nonconserved sequence were also identif ...19921452021
an inducible expression system for the production of human lactoferrin in aspergillus nidulans.the production and secretion of human lactoferrin (hlf) in aspergillus nidulans is described. the hlf cdna was expressed under the control of the strong ethanol-inducible alcohol dehydrogenase (alca) promoter. recombinant hlf (re-hlf) is produced at levels up to 5 micrograms/ml. approximately 30% of the re-hlf produced in this system is secreted into the growth medium. the re-hlf is indistinguishable from native hlf with respect to size and immunoreactivity. furthermore, re-hlf is functional by ...19921452033
cloning and molecular characterization of the glyceraldehyde-3-phosphate dehydrogenase-encoding gene and cdna from the plant pathogenic fungus glomerella cingulata.the glyceraldehyde-3-phosphate dehydrogenase gene (gpda) has been identified from a genomic dna library prepared from the plant pathogenic fungus glomerella cingulata. nucleotide sequence data revealed that this gene codes for a putative 338-amino-acid protein encoded by two exons of 129 and 885 bp, separated by an intron 216 bp long. the 5' leader sequence is also spliced by an intron of 156 bp. a cdna clone was prepared using the polymerase chain reaction, the sequence of which was used to con ...19921452034
purification, properties and primary structure of thioredoxin from aspergillus nidulans.this paper reports the purification and the properties of a thioredoxin from the fungus aspergillus nidulans. this thioredoxin is an acidic protein which exhibits an unusual fluorescence emission spectrum, characterized by a high contribution of tyrosine residues. thioredoxin from a. nidulans cannot serve as a substrate for escherichia coli thioredoxin reductase. corn nadp-malate dehydrogenase is activated by this thioredoxin in the presence of dithiothreitol, while fructose-1,6-bisphosphatase i ...19921459127
developmental and light regulation of eas, the structural gene for the rodlet protein of neurospora.the surface of many fungal spores is covered by a hydrophobic sheath termed the rodlet layer. we have determined that the rodlet protein of neurospora crassa is encoded by a cloned gene designated bli-7, and that bli-7 is identical to the known gene eas (easily wettable). using eas dna as a probe we show that eas mrna is abundant in illuminated mycelia and conidiophores but is not detectable or is barely detectable in dark-grown mycelia, mature macroconidia, microconidia, and ascospores. mutatio ...19921459459
the developmentally regulated aspergillus nidulans wa gene encodes a polypeptide homologous to polyketide and fatty acid synthases.the aspergillus nidulans wa gene is required for synthesis of a green pigment present in the walls of mature asexual spores (conidia); wa mutants produce colorless (white) conidia. we determined the transcriptional structure and dna sequence of the wa gene. wa consists of 5 exons separated by short (40-60 bp) introns. the processed transcript has the potential to encode a protein consisting of 1986 amino acid residues. the predicted wa polypeptide showed extensive sequence similarities with bact ...19921465094
characterization of the amda-regulated acia gene of aspergillus nidulans.the structure, function and regulation of the acetate inducible acia gene of aspergillus nidulans was analysed. the acia locus was mapped to chromosome 1 at a position where no acetate inducible gene has been previously located. the nucleotide sequence of acia was determined, the structures of two transcripts were determined and the sequences of the polypeptide products of the gene were deduced. construction of an acia loss-of-function mutant was achieved via insertional inactivation, but it did ...19921465107
identification of functional regions of the positively acting regulatory gene amdr from aspergillus nidulans.the amdr (inta) regulatory gene of aspergillus nidulans encodes a 765-amino-acid polypeptide which determines the omega-amino acid induction of at least five structural genes. the amdr polypeptide contains a potential zn(ii)2cys6 dna-binding motif which has been shown to be present in the n-terminal region of a large number of fungal activator proteins. in vitro mutagenesis of the fourth cysteine of this motif abolishes amdr function as shown by loss of complementation of an amdr- mutation and b ...19921479891
mitotic regulation in aspergillus nidulans.the nima and bime genes of aspergillus nidulans respectively encode a 79 kda protein kinase that is a positive regulator of mitosis and a 229 kda protein that is a negative regulator of mitosis. either overproduction of nima or inactivation of bime can induce mitosis and override the checkpoint associated with incomplete dna replication. double mutants between temperature-sensitive nima and bime alleles undergo chromatin condensation and spindle polymerization at restrictive temperature, suggest ...19921483343
isolation of an aspergillus terreus mutant impaired in arginine biosynthesis and its complementation with the argb gene from aspergillus nidulans.using filtration enrichment techniques, an aspergillus terreus arginine auxotrophic strain which contains a mutation that abolishes ornithine transcarbamylase (otcase) activity has been isolated. this mutant has been genetically transformed with the cloned aspergillus nidulans otcase gene. prototrophic transformants arose at a frequency of about 50 transformants per microgram of plasmid dna. southern blot analysis of dna from the transformants showed that the transforming dna was ectopically int ...19921490598
requirement for esp1 in the nuclear division of saccharomyces cerevisiae.mutations in the esp1 gene of saccharomyces cerevisiae disrupt normal cell-cycle control and cause many cells in a mutant population to accumulate extra spindle pole bodies. to determine the stage at which the esp1 gene product becomes essential for normal cell-cycle progression, synchronous cultures of esp1 mutant cells were exposed to the nonpermissive temperature for various periods of time. the mutant cells retained viability until the onset of mitosis, when their viability dropped markedly. ...19921493337
a dominant selectable marker that is meiotically stable in neurospora crassa: the amds gene of aspergillus nidulans.when neurospora crassa is transformed using a neurospora gene as the selectable marker, the vegetatively stable transformants obtained cannot be used successfully in a cross because the selectable marker will be inactivated by the process of rip (repeat-induced point mutation). introduction of the acetamidase-encoding gene amds of aspergillus nidulans into n. crassa by transformation yielded transformants that would grow in minimal medium containing acetamide as a sole nitrogen source. in mitoti ...19921494342
characterization of the aspergillus niger pelb gene: structure and regulation of expression.the nucleotide sequence of pelb, a member of the aspergillus niger pectin lyase multigene family, has been determined. the pelb gene product, plb, shares 65% amino acid identity with pectin lyase a (pla) and 60% with pectin lyase d (pld). although growth of pelb multicopy transformants on pectin-containing media results in elevated pelb mrna levels, pectin lyase b (plb) is barely detectable. this is probably due to degradation of plb by acid proteases, since multicopy transformants grown on pect ...19921495474
npk1, a nonessential protein kinase gene in saccharomyces cerevisiae with similarity to aspergillus nidulans nima.a new protein kinase gene [called npk1 (for nonessential protein kinase)] has been found on chromosome i of saccharomyces cerevisiae between cdc15 and ade1. the 435 amino acid/48 kda gene product is very similar to known protein kinases. it is most closely related to the nima protein of aspergillus nidulans, displaying 45.9% identity and 63.5% similarity in the protein kinase domain. a 1.4 kb transcript of the npk1 gene was detected. disruption of the npk1 gene impedes neither growth on glucose ...19921495480
arginine and proline genes of aspergillus niger.aspergillus niger mutants defective in arginine or proline biosynthesis have been isolated and 12 genetic loci were identified. mutation was induced by low doses uv, and mutants were isolated after filtration enrichment. the mutants were classified according to their phenotype in growth tests and were further characterized in complementation tests. the arginine auxotrophic mutants represent nine complementation groups. three additional complementation groups were found for mutants that could gro ...19921497330
early developmental events during asexual and sexual sporulation in aspergillus nidulans. 19921504597
control of gene expression in the catabolic pathways of aspergillus nidulans: a personal and biased account. 19921504607
protein synthesis in aspergillus nidulans.in this review of protein synthesis, we have described a system for translation of mrna using extracts of a. nidulans. this system is useful for characterizing mutants suspected to have defects in protein synthesis and for assessing the toxicity of various antibiotics and their effects on misreading the genetic code. the well developed genetical system of a. nidulans has enabled us to map at least 27 new genes whose mutation disturbs the level of translational accuracy. these mutants could be us ...19921504608
mutational analysis of calmodulin in saccharomyces cerevisiae.calmodulin is well characterized as an intracellular ca2+ receptor in nonproliferating tissues such as muscle and brain. several observations indicate that calmodulin is also required for cellular growth and division. deletion of the calmodulin gene is a lethal mutation in saccharomyces cerevisiae, schizosaccharomyces pombe and aspergillus nidulans. expression of calmodulin antisense rna in mouse c127 cells causes a transient arrest at g1 and metaphase. although these results indicate calmodulin ...19921505005
isolation of a gene required for programmed initiation of development by aspergillus nidulans.in contrast to many other cases in microbial development, aspergillus nidulans conidiophore production initiates primarily as a programmed part of the life cycle rather than as a response to nutrient deprivation. mutations in the acod locus result in "fluffy" colonies that appear to grow faster than the wild type and proliferate as undifferentiated masses of vegetative cells. we show that unlike wild-type strains, acod deletion mutants are unable to make conidiophores under optimal growth condit ...19921508186
the polygalacturonases of aspergillus niger are encoded by a family of diverged genes.aspergillus niger produces several polygalacturonases that, with other enzymes, are involved in the degradation of pectin. one of the two previously characterized genes coding for the abundant polygalacturonases i and ii (pgi and pgii) found in a commercial pectinase preparation was used as a probe to isolate five more genes by screening a genomic dna library in phage lambda embl4 using conditions of moderate stringency. the products of these genes were detected in the culture medium of aspergil ...19921511691
effect of restrictive conditions on the growth and morphology of a temperature-sensitive mannose-requiring mutant of aspergillus nidulans.when incubated at 45 degrees c in the absence of added mannose, pregrown hyphae of a temperature-sensitive, mannose-relief mutant (mnra455) of aspergillus nidulans grew normally for a short time (4-5 h) before exhibiting an abnormal morphology consisting of the production by hyphae of discrete spherical swellings called balloons. these swellings could be up to 10 microns in diameter and were produced either at or behind the hyphal apex. often only one swelling was produced in association with ea ...19921516805
stua is required for cell pattern formation in aspergillus.the stunted (stua) gene product is required for the orderly differentiation and spatial organization of cell types of the aspergillus nidulans conidiophore. expression of the stua gene is complex. two transcripts, stua alpha and stua beta, are initiated from separate promoters. transcription of both rnas increases approximately 50-fold during the establishment of developmental competence. induction-dependent transcriptional and post-transcriptional regulatory mechanisms further enhance expressio ...19921516832
chromatin structure of schizosaccharomyces pombe. a nucleosome repeat length that is shorter than the chromatosomal dna length.we have used new methods for chromatin isolation, together with conventional methods for measuring the nucleosome repeat length, to determine the repeat length of schizosaccharomyces pombe chromatin. we obtain a result of 156(+/- 2) bp. equivalent results are obtained using a psoralen crosslinking method for measuring the repeat length in viable spheroplasts. that result, together with other control experiments, rules out many possible artifacts. the measured value of 156(+/- 2) bp is smaller th ...19921518041
phase i study of yk-176 (2'-deoxycoformycin) in patients with adult t-cell leukemia-lymphoma. the dcf study group.yk-176 is a newly isolated 2'-deoxycoformycin (dcf), a potent inhibitor of adenosine deaminase, produced by aspergillus nidulans. in a cooperative phase i study, yk-176 was administered to 22 patients, comprising 18 with adult t-cell leukemia-lymphoma (atl), two with cutaneous t-cell lymphoma (ctcl), one with lymphoblastic lymphoma of t-cell type and one with carcinoma of the uterine cervix. doses of yk-176 ranged from 3.0 to 9.0 mg/m2 and were given intravenously for three consecutive days. gen ...19921518164
protein structure of pig liver 4-aminobutyrate aminotransferase and comparison with a cdna-deduced sequence.the amino acid sequence of pig liver 4-aminobutyrate aminotransferase has been determined by gas-phase sequencing of proteolytically derived peptide fragments. the sequence differs substantially from that predicted for the same enzyme on the basis of the sequence of cdna derived from pig brain in recently published work [kwon, o., park, j. & churchich, j. e. (1992) j. biol. chem. 267, 7215-7216]. apart from a few minor differences, the two sequences are completely different in the segment of pro ...19921521531
isolation and characterization of an analogue-resistant aminoacyl-trna synthetase mutant in aspergillus nidulans.six mutants resistant to p-fluorophenylalanine (fpa) were selected on a medium containing aspartate as the sole source of nitrogen using a phenylalanine-requiring (phena)auxotroph of a. nidulans as the wild type. the mutants, on the basis of genetic characterization, were found to be alleilic and located on the left arm of the linkage group iii, approximately 13 map unit left to meth h locus, henceforth assigned to the symbol fpav. at a fixed concentration of phenylalanine (23 micrograms/ml), th ...19921521872
[influence of heavy metal ions on the electrophysical properties of anacystis nidulans and escherichia coli cells].the influence of heavy metal ions (ag+, cu2+, cd2+, pb2+, mn2+, zn2+, gd3+, 1 microm-1 mm) on anacystis nidulans and escherichia coli cells has been studied by means of electrophoresis and electro-orientation spectroscopy methods. it has been shown that changes of cell electrophoretic mobility (em) and low-frequency (20 hz) electro-orientation effect (eoe) observed with the increase of metal cation concentration characterize the adsorption of these ions on surface layers of cell envelopes. the d ...19921522819
binding selectivity of rhizoxin, phomopsin a, vinblastine, and ansamitocin p-3 to fungal tubulins: differential interactions of these antimitotic agents with brain and fungal tubulins.the binding of four potent antimitotic agents, rhizoxin (rzx), phomopsin a (pms-a), ansamitocin p-3 (asmp-3), and vinblastine (vlb), to tubulins from rzx-sensitive and -resistant strains of aspergillus nidulans, schizosaccharomyces pombe, and saccharomyces cerevisiae was investigated. mycelial extracts to which rzx could bind contained beta-tubulin with asn as the 100th amino acid residue (asn-100) in all cases, and those without affinity for rzx contained beta-tubulins with either ile-100 or va ...19921530630
characterization of the glyoxysomal isocitrate lyase genes of aspergillus nidulans (acud) and neurospora crassa (acu-3).the nucleotide sequences of the genes encoding the acetate-inducible glyoxylate cycle enzyme isocitrate lyase from the ascomycete fungi aspergillus nidulans (acud) and neurospora crassa (acu-3) are presented. the respective a. nidulans and n. crassa genes are interrupted at identical positions by two introns and encode proteins of 538 and 543 amino acids, which have 75% identity. the predicted protein sequences do not demonstrate the c-terminal tripeptide s-k-l that has been implicated in peroxi ...19921531185
starch digestion and adsorption by beta-amylase of emericella nidulans (aspergillus nidulans).a mutant strain of emericella nidulans mnu 82 was isolated by multistep mutation. the beta-amylase produced by the mutant was able to digest raw starch. it was readily and strongly adsorbed onto raw starch at ph 5.0. the enzyme to starch ratio was 1950 u/g starch. the enzyme showed no correlation between the capacity of raw starch digestion and adsorption of the enzyme.19921533208
an extra copy of nimecyclinb elevates pre-mpf levels and partially suppresses mutation of nimtcdc25 in aspergillus nidulans.previous work has shown that nima encodes a cell cycle regulated protein kinase that is required along with the p34cdc2 histone h1 kinase (mpf) for mitosis in aspergillus nidulans. we have now identified two other gene products required for mitosis in a.nidulans. nimt encodes a protein similar to the fission yeast cdc25 tyrosine phosphatase and is required for the conversion of pre-mpf to mpf and nime encodes a b-type cyclin which is a subunit of mpf. a new genetic interaction between nimecyclin ...19921534750
nitrate reductase and nitrite reductase transcript levels in various mutants of aspergillus nidulans: confirmation of autogenous regulation.the regulation of the expression of the a. nidulans niia and niad genes (encoding nitrite reductase and nitrate reductase activities, respectively) was investigated by northern blotting. it was demonstrated that expression of the niia and niad genes is controlled at the level of mrna accumulation and that mutations within the nira and area regulatory genes, as well as certain mutations within niad itself or cnxe (for its molybdenum cofactor), markedly affect niia and niad transcript levels.19921538701
kinesin-related cut7 protein associates with mitotic and meiotic spindles in fission yeast.several mitotic and meiotic gene products are related to the microtubule motor kinesin, providing insight into the molecular basis of the complex motile events responsible for spindle formation and function. of these genes, three have been shown to affect spindle structure when mutated. the most severe phenotype is seen in aspergillus nidulans bimc and schizosaccharomyces pombe cut7 mutants. in both fungi the intranuclear spindle is bipolar, with microtubules that emanate from spindle pole bodie ...19921538784
gamma-tubulin is a centrosomal protein required for cell cycle-dependent microtubule nucleation.gamma-tubulin is a newly identified member of the tubulin family whose sequence is highly conserved from yeast to man. this minor microtubule protein is localized to the microtubule organizing centres and a mutation in the gene encoding it produces a microtubuleless mitotic arrest in the filamentous fungus aspergillus nidulans. here we investigate the in vivo function of gamma-tubulin in mammalian cells using a synthetic peptide to generate a polyclonal antibody that binds to a highly conserved ...19921538786
the aspergillus niger niad gene encoding nitrate reductase: upstream nucleotide and amino acid sequence comparisons.the aspergillus niger niad gene has been sequenced and the inferred nitrate reductase (nr) protein found to consist of 867 amino acid residues (97 kda). the gene is interrupted by six small introns, as deduced by comparison with the niad gene of aspergillus nidulans. the positions of these putative introns are conserved between the two fungi, although the sequences are dissimilar. the niia gene, encoding nitrite reductase, the second reductive step in the nitrate assimilation pathway, is tightly ...19921541396
inducible alkyltransferase dna repair proteins in the filamentous fungus aspergillus nidulans.we have investigated the response of the filamentous fungus aspergillus nidulans to low, non-killing, doses of the alkylating agent mnng (n-methyl-n'-nitro-n-nitrosoguanidine). such treatment causes a substantial induction of dna alkyltransferase activity, with the specific activity in treated cells increasing up to one hundred-fold. fluorography reveals the two main inducible species as proteins of 18.5 kda and 21 kda, both of which have activity primarily against o6-methylguanine (o6-meg) lesi ...19921542560
structure of the aspergillus nidulans qut repressor-encoding gene: implications for the regulation of transcription initiation.the nucleotide (nt) sequence of the qutr gene has been determined and shown to encode an inferred protein (qutr) of 929 amino acids (aa). the inferred aa sequence shows a high level of similarity throughout its length with the aa sequence of the three c-terminal domains (shikimate kinase; 3-dehydroquinase; shikimate dehydrogenase) of the pentafunctional arom protein of aspergillus nidulans that catalyses steps 2-6 in the shikimate pathway. the inferred qutr aa sequence has a completely conserved ...19921544567
transfer of isolated nuclei into protoplasts of aspergillus nidulans.nuclei were isolated from protoplasts of a haploid auxotrophic aspergillus nidulans strain. transformation of protoplasts prepared from a complementary haploid auxotrophic strain with these purified nuclei resulted in both heterokaryotic and diploid colonies. the nutritionally-complementing colonies appeared at a frequency of 5 x 10(-7) to 10(-8).19921547859
a comparison of the enzymological and biophysical properties of two distinct classes of dehydroquinase enzymes.this paper compares the biophysical and mechanistic properties of a typical type i dehydroquinase (dhqase), from the biosynthetic shikimate pathway of escherichia coli, and a typical type ii dhqase, from the quinate pathway of aspergillus nidulans. c.d. shows that the two proteins have different secondary-structure compositions; the type i enzyme contains approx. 50% alpha-helix while the type ii enzyme contains approx. 75% alpha-helix. the stability of the two types of dhqase was compared by de ...19921554351
protein kinases from aspergillus nidulans that phosphorylate the carboxyl-terminal domain of the largest subunit of rna polymerase ii.three serine kinases which phosphorylate the ctd of rna polymerase ii have been identified in aspergillus nidulans. the kinases (ki, kii, kiii) were identified using a synthetic peptide containing four copies of the ctd consensus heptamer repeat, and differ in chromatographic behavior, and apparent molecular mass (ki approximately 60kda; kii approximately 82kda; kiii approximately 43 kda). kiii utilized, in addition to peptide, histone h1 as substrate, whereas casein was not phosphorylated by an ...19921556138
disruption of a magnaporthe grisea cutinase gene.using a one-step strategy to disrupt cut1, a gene for cutinase, cut1- mutants were generated in two strains of magnaporthe grisea. one strain, pathogenic on weeping lovegrass and barley and containing the arg3-12 mutation, was transformed with a disruption vector in which the aspergillus nidulans argb+ gene was inserted into cut1. prototrophic transformants were screened by southern hybridization, and 3 of 53 tested contained a disrupted cut1 gene (cut1::argb+). a second strain, pathogenic on ri ...19921557024
sequence analysis of the gene coding for glyceraldehyde-3-phosphate dehydrogenase (gpd) of podospora anserina: use of homologous regulatory sequences to improve transformation efficiency.the glyceraldehyde-3-phosphate dehydrogenase (gpd) gene of podospora anserina has been isolated from a genomic library by heterologous hybridization with the corresponding gene of curvularia lunata. the coding region consists of 1014 nucleotides and is interrupted by a single intron. the amino-acid sequence encoded by the gpd gene shows a high degree of sequence identity with the corresponding gene products of various fungi. multiple alignments of all fungal gpd sequences so far available result ...19921563046
cloning and sequence determination of a cdna encoding aspergillus nidulans calmodulin-dependent multifunctional protein kinase.a partial cdna encoding aspergillus nidulans calmodulin-dependent multifunctional protein kinase (acmpk) was isolated from a lambda zap expression library by immunoselection using monospecific polyclonal antibodies to the enzyme. the sequence of both strands of the cdna (cmka) was determined. the deduced amino acid (aa) sequence contained all eleven consensus domains found in serine/threonine protein kinases [hanks et al., science 241 (1988) 42-52], as well as a putative calmodulin-binding domai ...19921563634
identification of the promoter region involved in the autoregulation of the transcriptional activator alcr in aspergillus nidulans.the alcr protein is the transcriptional activator of the ethanol utilization pathway in the filamentous fungus aspergillus nidulans. this activator belongs to a family of fungal proteins having a conserved dna-binding domain containing six cysteines (c6 class) with some striking features. at variance with other motifs of this class, the binding domain of alcr is strongly asymmetrical in relation to the central cysteines and moreover was predicted to adopt a helix-turn-helix structure. this domai ...19921569930
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