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genesis of eukaryotic transcriptional activator and repressor proteins by splitting a multidomain anabolic enzyme.the genes necessary for the correctly regulated catabolism of quinate in aspergillus nidulans and neurospora crassa are controlled at the level of transcription by a dna-binding activator protein and a repressor protein that directly interact with one another. the repressor protein is homologous throughout its length with the three c-terminal domains of a pentafunctional enzyme catalysing five consecutive steps in the related anabolic shikimate pathway. we now report that the activator protein i ...19938294040
pclk1 and pclt5--two linear mitochondrial plasmids from unrelated claviceps purpurea strains: a comparison.pclt5, a linear mitochondrial (mt) plasmid from claviceps purpurea, strain t5, was sequenced and compared to pclk1, a linear mt plasmid from an unrelated c. purpurea strain. both plasmids have terminal proteins (tps) at their inverted terminal repeats (tlr). the tlrs of both plasmids show short conserved sequences, which are probably involved in plasmid transcription and replication. the coding capacity of pclt5 and pclk1 is similar: there are two large orfs (orf1 and orf2) homologous to the dna ...19938302935
two binding sites of inhibitors in nadh: ubiquinone oxidoreductase (complex i). relationship of one site with the ubiquinone-binding site of bacterial glucose:ubiquinone oxidoreductase.the effect of ten naturally occurring and two synthetic inhibitors of nadh:ubiquinone oxidoreductase (complex i) of bovine heart, neurospora crassa and escherichia coli and glucose:ubiquinone oxidoreductase (glucose dehydrogenase) of gluconobacter oxidans was investigated. these inhibitors could be divided into two classes with regard to their specificity and mode of action. class i inhibitors, including the naturally occurring piericidin a, annonin vi, phenalamid a2, aurachins a and b, thiangaz ...19948307034
calcium and microorganisms.this review followed from experiments suggesting that some fungi do not require calcium. it was found that many studies of a calcium requirement in microorganisms had assumed specificity for chelation agents such as egta and a23187, which the reagents did not possess. early studies still cited today often preceded the recognition that microorganisms required manganese and zinc. as a result of both of these misunderstandings, there was rarely any attempt to replace calcium by other important trac ...19938338620
the positive-acting sulfur regulatory protein cys3 of neurospora crassa: nuclear localization, autogenous control, and regions required for transcriptional activation.the positive-acting global sulfur regulatory protein, cys3, of neurospora crassa turns on the expression of a family of unlinked structural genes that encode enzymes of sulfur catabolism. cys3 contains a leucine zipper and an adjacent basic region (b-zip), which together constitute a bipartite sequence-specific dna-binding domain. specific anti-cys3 antibodies detected a protein of the expected size in nuclear extracts of wild-type neurospora under conditions in which the sulfur circuit is activ ...19938316209
identification of a g protein alpha subunit from neurospora crassa that is a member of the gi family.heterotrimeric g proteins, consisting of alpha, beta, and gamma subunits, are implicated in major signal transduction pathways controlling a diversity of functions in eukaryotic organisms. in the filamentous fungus neurospora crassa, g proteins are implicated in the regulation of several environmental responses. as a first step in studying the role of g proteins in these processes, we have cloned the genes for two alpha subunits, gna-1 and gna-2, from neurospora. the genes are located on differe ...19938325859
heterologous expression of mating-type genes in filamentous fungi.podospora anserina and neurospora crassa, two filamentous heterothallic ascomycetes, have a single mating-type locus with two alternate forms called mat+ and mat- and a and a, respectively. mating type controls entry into the sexual cycle, events subsequent to fertilization, and, in n. crassa, prevents the formation of mixed mating-type heterokaryons. the mating types of these two organisms display similarity in their dna structure and in the encoded polypeptides involved in fertilization. here ...19938341677
is an activator protein-2-like transcription factor involved in regulating gene expression during nitrogen limitation in fungi?the upstream sequences of all published lignin peroxidase and manganese peroxidase genomic clones from phanerochaete chrysosporium were analyzed. this analysis revealed the presence of putative activator protein-2 (ap-2) recognition sequences in 11 of 15 lignin peroxidase genes. the lignin peroxidase clone glg6 and the manganese peroxidase gene (mnp-1) have two copies of putative ap-2 sequence in the upstream region. interestingly, the lignin peroxidase gene vlg4 of another white rot fungus, tra ...19938357266
choline: its role in the growth of filamentous fungi and the regulation of mycelial morphology.choline is an essential metabolite for the growth of filamentous fungi. it occurs most notably as a component of the major membrane phospholipid, phosphatidyl choline (lecithin), and fulfills a major role in sulphate metabolism in the form of choline-o-sulphate in many species. choline is usually synthesised endogenously, but exogenous choline can also be taken up, either to compensate for metabolic deficiencies in choline-requiring mutants such as those of aspergillus nidulans and neurospora cr ...19938318261
cloning of the gene for glutamate decarboxylase and its expression during conidiation in neurospora crassa.neurospora crassa glutamate decarboxylase (gad) is produced during conidiation and stored in dormant conidia. polyclonal antibody was generated to gad that had been purified to homogeneity. the anti-gad antibody was specific for n. crassa gad and inhibited gad activity. the level of gad protein decreased during conidial germination, indicating that gad was degraded during this phase of development. the anti-gad antibody was used to isolate a cdna clone of gad from a lambda zap cdna expression li ...19938352739
the sta2 and mel1 genes of saccharomyces cerevisiae are idiomorphic.the sta2 (glucoamylase) gene of saccharomyces cerevisiae has been mapped close to the end of the left arm of chromosome ii. meiotic analysis of a cross between a haploid strain containing sta2, and another strain carrying the melibiase gene mel1 (which is known to be at the end of the left arm of chromosome ii) produced parental ditype tetrads only. since there is no significant dna sequence similarity between the sta2 and mel1 genes, or their respective flanking regions, we conclude that these ...19938381338
functional domains of the transcriptional activator nuc-1 in neurospora crassa.the nuc-1 regulatory protein directly controls the transcription of these genes and how the activity enzymes in neurospora crassa. to understand how nuc-1 regulates the transcription of these genes and how the activity of nuc-1 is modulated by other regulatory proteins, two putative functional domains of nuc-1 were analysed: the dna-binding domain and the regulatory domain. the dna-binding activity of nuc-1 has not been directly demonstrated; however, results of deletion analysis, sequence analy ...19938359693
functional identification of al-3 from neurospora crassa as the gene for geranylgeranyl pyrophosphate synthase by complementation with crt genes, in vitro characterization of the gene product and mutant analysis.in this work the neurospora crassa al-3 gene function was determined. geranylgeranyl pyrophosphate (ggpp) synthase activity was measured in al-2 fgsc 313 and al-3 rp100 fgsc 2082 mutant strains by in vitro synthesis methods. this experiment showed that al-3 rp100 mutant expresses a reduced ggpp synthase activity. the mutated al-3 gene was cloned and sequenced; a single missense mutation was found changing serine into asparagine. genetic complementation was performed by escherichia coli transform ...19938350190
purification and characterization of the assimilatory nitrate reductase of azotobacter vinelandii.1. a soluble reduced methyl viologen-dependent assimilatory nitrate reductase from azotobacter vinelandii strain uw136 grown aerobically on nitrate was purified to homogeneity by the criteria of nitrate reductase activity staining, and coincidence of a coomassie blue-staining protein band on polyacrylamide gels run under non-denaturing conditions. the specific activity was 3 mumol of no2- formed/min per mg of protein. 2. gel filtration on superose-12 and sds/page showed that the enzyme had an m( ...19938380991
interchangeable associations of calcineurin regulatory subunit isoforms with mammalian and fungal catalytic subunits.two mammalian genes for the ca(2+)-binding regulatory subunit of the calmodulin-dependent protein phosphatase (calcineurin) were identified recently, suggesting unique associations with tissue-specific catalytic subunits. the murine brain (beta 1) and testis (beta 2) isoforms of the regulatory subunit were expressed with poly-histidine carboxyl termini and purified by ni(2+)-chelate chromatography, each exhibiting high affinity ca2+ binding on nitrocellulose overlays. using chromatographic metho ...19938384215
mitochondrial cardiolipin in diverse eukaryotes. comparison of biosynthetic reactions and molecular acyl species.cardiolipin, a unique dimeric phospholipid of bacteria and mitochondria, can be synthesized by two alternative pathways discovered in rat and escherichia coli, respectively. in mitochondrial preparations from fungi (saccharomyces cerevisiae, neurospora crassa), higher plants (phaseolus aureus), molluscs (mytilus edulis) and mammals (rat liver, bovine adrenal gland), cardiolipin was synthesized from cdp-diacylglycerol and phosphatidylglycerol, suggesting a common eukaryotic mechanism of cardiolip ...19938385010
multiple isomers of phosphatidyl inositol monophosphate and inositol bis- and trisphosphates from filamentous fungi.the range of inositol phosphates and inositol phospholipids present in three filamentous fungi, neurospora crassa, fusarium graminearum and phanerochaete chrysosporium has been investigated by hplc analysis. the profiles obtained demonstrate that two isomers of phosphatidyl inositol monophosphate are present, and that an apparent complexity in the number of isomers of inositol bis- and trisphosphates is found in filamentous fungi that has not been observed in animal or plant cells.19938394259
glycogen-bound protein in lower eukaryote and prokaryote.the proteoglycogen fraction of neurospora crassa was purified and subjected to radioiodination with [125i]iodide. amylolysis of the polysaccharide moiety led to the isolation of a labelled 31 kda-protein. the nh2-terminal amino acid sequence of 10 residues of the 31 kda-protein was determined. a 31 kda-protein was also bound to glycogen in escherichia coli. proteoglycogen has not been heretofore found in any primitive unicellular organism.19938401303
complete coding sequence, intron/exon organization, and chromosomal location of the gene for the core i protein of human ubiquinol-cytochrome c reductase.core i protein is a nuclear-encoded component of the ubiquinol-cytochrome c reductase complex of the mitochondrial respiratory chain. we have located the gene for the human core i protein in the p21 region of chromosome 3, just upstream of the col7a1 gene which encodes type vii collagen. the core i gene, which has been sequenced in its entirety, is comprised of 10,417 base pairs, from the major transcription start site to the polyadenylation signal, and contains 13 exons. the predicted polypepti ...19938407948
a tobacco cdna clone encoding a gata-1 zinc finger protein homologous to regulators of nitrogen metabolism in fungi.in higher plants, the expression of the nitrate assimilation pathway is highly regulated. although the molecular mechanisms involved in this regulation are currently being elucidated, very little is known about the trans-acting factors that allow expression of the nitrate and nitrite reductase genes which code for the first enzymes in the pathway. in the fungus neurospora crassa, nit-2, the major nitrogen regulatory gene, activates the expression of unlinked structural genes that specify nitroge ...19938413186
primary structure of the nuclear-encoded 10.5 kda subunit of complex i from neurospora crassa.we have isolated a cdna clone for the nuclear encoded 10.5 kda subunit of complex i from n. crassa. dna sequencing revealed an open reading frame corresponding to a polypeptide with 94 amino acids and a calculated molecular mass of 10531 da. the protein is synthesized without a cleavable mitochondrial targeting sequence. the n. crassa polypeptide is the fungal equivalent of subunit b8 of bovine complex i.19938448210
molecular cloning of the smaller subunit(p52) of rat liver mitochondrial processing protease.a cdna encoding the smaller subunit (p52) of mitochondrial processing protease was isolated from a rat liver cdna library, using cdna fragment for yeast mas1 as the probe. the deduced amino acid sequence is highly homologous to those of pep from neurospora crassa and mas1 from saccharomyces cerevisiae. after in vitro transcription and translation, the precursor peptide was imported into isolated rat liver mitochondria and processed to its mature form.19938422255
insertional mutagenesis in neurospora crassa: cloning and molecular analysis of the preg+ gene controlling the activity of the transcriptional activator nuc-1.the transcriptional activator nuc-1 controls the transcription of the genes for phosphorus acquisition enzymes, and its activity is regulated by the negative regulatory factors, preg and pgov in this report, we describe the cloning and molecular analysis of the preg+ gene. in neurospora crassa, as in higher eukaryotes, transformation frequently results in nonhomologous integration of transforming dna. insertion of transforming dna into host genes mutates the gene and provides a molecular tag for ...19938436269
the mitochondrial genome of the entomopathogenic fungus beauveria bassiana: analysis of the ribosomal rna region.the 28.5-kbp mitochondrial (mt) genome from the entomopathogenic fungus beauveria bassiana was studied using restriction enzyme analysis, gene probe hybridization, and dna sequence comparisons. a detailed restriction enzyme map allowed cloning of the entire genome into a number of segments. hybridization of heterologous gene probes to the mtdna resulted in the identification of the large ribosomal rna (lrrna) and small ribosomal rna (srrna) genes. gene probes derived from several yeasts and fung ...19938439871
peptide splicing in the vacuolar atpase subunit a from candida tropicalis.subunit a of the vacuolar proton pump appears to be responsible for the atp hydrolysis which is coupled to the pumping of protons into a variety of intracellular acid compartments, including the fungal vacuole. we report here the cloning and sequence determination of the gene encoding subunit a from candida tropicalis. southern blot hybridization analysis indicates that there is a single gene which encodes this protein. the gene contains a single intron at the extreme 5'-end of the coding region ...19938463270
isolation, sequencing, and characterization of crp-5, a gene encoding a neurospora ribosomal protein.a neurospora crassa cytoplasmic ribosomal protein gene, crp-5, has been isolated and characterized. the cdna was isolated by a differential screening of a cdna library for glucose-inducible genes. the cdna was subsequently used to identify and isolate crp-5 genomic sequences. computer analysis of the dna sequences showed that they contain an open reading frame which encodes a protein homologous to the rat ribosomal protein s26. the crp-5 mrna levels are regulated in a carbon-source-dependent man ...19938467530
cloning and analysis of the mating type genes from cochliobolus heterostrophus.cochliobolus heterostrophus, a heterothallic ascomycete, has a single mating type locus with two alternate forms called mat-1 and mat-2. mat-1 was cloned by complementing a mat-2 strain using a cosmid library from a mat-1 strain and screening for a homothallic transformant. the cosmid recovered from this transformant was able to re-transform a mat-2 strain to homothallism and mat identity was proven by restriction fragment length polymorphism and conventional genetic mapping. all homothallic tra ...19938479433
developmental regulation of the gene for formate dehydrogenase in neurospora crassa.we have isolated and characterized a gene, fdh, from neurospora crassa which is developmentally regulated and which produces formate dehydrogenase activity when expressed in escherichia coli. the gene is closely linked (less than 0.6 kb apart) to the leu-5 gene encoding mitochondrial leucyl-trna synthetase; the two genes are transcribed convergently from opposite strands. the expression patterns of these genes differ: fdh mrna is found only during conidiation and early germination and is not det ...19938509325
cloning and sequencing of the genes coding for the a and b subunits of vacuolar-type na(+)-atpase from enterococcus hirae. coexistence of vacuolar- and f0f1-type atpases in one bacterial cell.the eubacterium enterococcus hirae atcc 9790 possesses a h(+)-translocating atpase, and the deduced amino acid sequences of the genes coding for this enzyme have indicated that it is a typical f0f1-type atpase (shibata, c., ehara, t., tomura, k., igarashi, k., and kobayashi, h. (1992) j. bacteriol. 174, 6117-6124). we cloned the ntpa and ntpb genes coding for the a and b subunits, respectively, of na(+)-translocating atpase from the same bacterium, and the full amino acid sequences of the two su ...19938505293
deletion mutations in the speed operon: spermidine is not essential for the growth of escherichia coli.null mutants of escherichia coli were constructed that cannot synthesize spermidine, because of deletions in the gene encoding s-adenosylmethionine decarboxylase. these mutants are still able to grow at near normal rates in purified media deficient in polyamines. these results in e. coli differ from recent findings that null mutants of saccharomyces cerevisiae and of neurospora crassa have an absolute growth requirement for spermidine.19938472951
phylogenetic analysis of ten black yeast species using nuclear small subunit rrna gene sequences.the nuclear small subunit rrna genes of authentic strains of the black yeasts exophiala dermatitidis, wangiella dermatitidis, sarcinomyces phaemuriformis, capronia mansonii, nadsoniella nigra var. hesuelica, phaeoannellomyces elegans, phaeococcomyces exophialae, exophiala jeanselmei var. jeanselmei and e. castellanii were amplified by pcr and directly sequenced. a putative secondary structure of the nuclear small subunit rrna of exophiala dermatitidis was predicted from the sequence data. alignm ...19958526477
isolation and characterization of the catalytic subunit of protein phosphatase 2a from neurospora crassa.the catalytic subunit of protein phosphatase 2a (pp2ac) was purified from neurospora crassa extract by (nh4)2so4-ethanol precipitation followed by deae-sephacel, heparin-sepharose, and monoq chromatography steps about 900-fold to a specific activity of 1200 u/g with a 2% yield. the apparent m(r) of pp2ac was estimated to be 35 kda by gel filtration and 33 kda by sds polyacrylamide gel electrophoresis. half maximal inhibition of pp2ac was achieved at 0.3 nm okadaic acid, 0.1 nm microcystin-lr, 56 ...19958529028
the beta subunit of the mitochondrial processing peptidase from rat liver: cloning and sequencing of a cdna and comparison with a proposed family of metallopeptidases.most nuclearly encoded mitochondrial proteins are synthesized with amino-terminal leader peptides that are removed by the mitochondrial processing peptidase (mpp) after translocation. earlier we reported cloning and sequencing of a cdna for the larger subunit (mpp alpha subunit) of this enzyme from rat liver mitochondria. we have now completed the cloning and sequencing of a cdna encoding the smaller subunit of the enzyme (mpp beta subunit) from the same source. the cdna consists of 1570 bp: 17 ...19938506385
site-directed mutagenesis of the cysteine residues in the neurospora crassa plasma membrane h(+)-atpase.a high-yield yeast expression system for site-directed mutagenesis of the neurospora crassa plasma membrane h(+)-atpase has recently been reported (mahanty, s. k., rao, u. s., nicholas, r. a., and scarborough, g. a. (1994) j. biol. chem. 269, 17705-17712). using this system, each of the eight cysteine residues in the atpase was changed to a serine or an alanine residue, producing strains c148s and c148a, c376s and c376a, c409s and c409a, c472s and c472a, c532s and c532a, c545s and c545a, c840s a ...19968550588
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
the roles of ca2+ and plasma membrane ion channels in hyphal tip growth of neurospora crassa.growing hyphae of the ascomycete fungus neurospora crassa contained a tip-high gradient of cytoplasmic ca2+, which was absent in non-growing hyphae and was insensitive to gd3+ in the medium. patch clamp recordings in the cell-attached mode, from the plasma membrane of these hyphae, showed two types of channel activities; spontaneous and stretch activated. the spontaneous channels were identified as inward k+ channels based on inhibition by tetraethylammonium. the stretch activated channels had i ...19958586653
identification and expression of the neurospora crassa mei-3 gene which encodes a protein homologous to rad51 of saccharomyces cerevisiae.the mei-3 gene of neurospora crassa encodes a homolog of the escherichia coli reca and saccharomyces cerevisiae rad51 proteins, which are required for recombination and repair of dna double-strand breaks. to determine the molecular function of mei3 protein, anti-mei3 antibody was prepared and used in western blot analysis. the antibody cross-reacted only with crude extracts prepared from perithecia, the fruiting bodies of neurospora. the molecular weight of the mei3 protein was estimated to be 3 ...19958552049
functional analysis by site-directed mutagenesis of individual amino acid residues in the flavin domain of neurospora crassa nitrate reductase.nitrate reductase of neurospora crassa is a complex multi-redox protein composed of two identical subunits, each of which contains three distinct domains, an amino-terminal domain that contains a molybdopterin cofactor, a central heme-containing domain, and a carboxy-terminal domain which binds a flavin and a pyridine nucleotide cofactor. the flavin domain of nitrate reductase appears to have structural and functional similarity to ferredoxin nadph reductase (fnr). using the crystal structure of ...19958552051
detection of additional restriction fragment length polymorphisms among the weakly virulent (nonaggressive) and highly virulent (aggressive) isolates of leptosphaeria maculans.isolates of leptosphaeria maculans were analyzed for their genetic relatedness based on dna restriction fragment length polymorphisms (rflps), employing as southern hybridization probes a combination of heat shock responsive genes (hsp70 and hsp80 from neurospora crassa), the cutinase gene of magnaporthe grisea, and cloned genomic dna sequences from a virulent strain. southern hybridization analysis revealed a high frequency of dna polymorphism. restriction fragments generated by each enzyme-pro ...19958542555
the novel nuclear gene dss-1 of saccharomyces cerevisiae is necessary for mitochondrial biogenesis.a previously unknown nuclear gene dss-1 from saccharomyces cerevisiae was cloned and sequenced. the gene was isolated as a multicopy suppressor of a disruption of the suv-3 gene coding for a dead/h box protein involved in processing and turnover of mitochondrial transcripts. the dss-1 gene codes for a 970 amino-acid protein of molecular weight 111 kda and is necessary for mitochondrial biogenesis. amino-acid sequence analysis indicates the presence of motifs characteristic for escherichia coli r ...19958590460
expression of the neurospora crassa metallothionein gene in escherichia coli and its effect on heavy-metal uptake.the gene coding for the neurospora crassa metallothionein protein was chemically synthesized and cloned into the fusion expression vectors pmal-c and pmal-p. cell-fractionation experiments demonstrated the proper localization of the pmal-c and pmal-p- expressed proteins to the cytosol and periplasm of the bacteria respectively. control bacteria as well as the recombinant bacteria producing the metallothionein protein were incubated with solutions of 109cd at concentrations of 0.2 microm, 1 micro ...19958590662
the orientation of gene maps by recombination of flanking markers for the am locus of neurospora crassa.fincham (1967), smyth (1973 b) and rambosek and kinsey (1983) have each generated fine-structure maps of the am gene of neurospora crassa. each map had a consistent linear order of alleles but the assignment of an orientation with respect to other linkage group-v loci differed. fincham found the end marked by the am6 allele to be at the distal end of the locus, smyth found am6 to be at the proximal end while the data of rambosek and kinsey did not suggest an orientation. smyth s orientation has ...19958595655
different respiratory-defective phenotypes of neurospora crassa and saccharomyces cerevisiae after inactivation of the gene encoding the mitochondrial acyl carrier protein.the nuclear genes (acp-1, acp1) encoding the mitochondrial acyl carrier protein were disrupted in neurospora crassa and saccharomyces cerevisiae. in n. crassa acp-1 is a peripheral subunit of the respiratory nadh : ubiquinone oxidoreductase (complex i). s. cerevisiae lacks complex i and its acp1 appears to be located in the mitochondrial matrix. the loss of acp-1 in n. crassa causes two biochemical lesions. firstly, the peripheral part of complex i is not assembled, and the membrane part is not ...19958595652
mitochondrial receptor complex from neurospora crassa and saccharomyces cerevisiae. 19958592451
generation and characterization of nadh: ubiquinone oxidoreductase mutants in neurospora crassa. 19958592454
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
mutational analysis reveals dispensability of the n-terminal region of the aspergillus transcription factor mediating nitrogen metabolite repression.mutational analysis has enabled identification and localization of an upstream exon of the area gene of aspergillus nidulans mediating nitrogen metabolite repression. a mutation in the initiation codon and frameshift mutations, which revert by restoration of the reading frame, established the coding role of the exon and mutations affecting intron splicing in conjunction with dna sequencing of reverse transcriptase polymerase chain reaction (rt-pcr) products localized the coding region intron. th ...19958596437
functional molecular aspects of the nadh dehydrogenases of plant mitochondria.there are multiple routes of nad(p)h oxidation associated with the inner membrane of plant mitochondria. these are the phosphorylating nadh dehydrogenase, otherwise known as complex i, and at least four other nonphosphorylating nad(p)h dehydrogenases. complex i has been isolated from beetroot, broad bean, and potato mitochondria. it has at least 32 polypeptides associated with it, contains fmn as its prosthetic group, and the purified enzyme is sensitive to inhibition by rotenone. in terms of su ...19958595975
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
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
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
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
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
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
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
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
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
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
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
fungal membrane responses induced by plant defensins and thionins.treatment of hyphae of neurospora crassa with antifungal plant defensins, i.e. rs-afp2 and dm-amp1 isolated from radish and dahlia seed, respectively, induced a rapid k+ efflux, ca2+ uptake, and alkalinization of the incubation medium. the rs-afp2-induced alkalinization of the incubation medium could be inhibited with g-protein inhibitors. alpha-hordothionin, an antifungal thionin from barley seed, caused a sustained increased ca2+ uptake at subinhibitory concentrations but only a transient incr ...19968663029
the role of the n and c termini of recombinant neurospora mitochondrial porin in channel formation and voltage-dependent gating.to investigate the role of the n and c termini in channel function and voltage-dependent gating of mitochondrial porin, we expressed wild-type and mutant porins from neurospora crassa as his-tag fusion products in escherichia coli. large quantities of the proteins were purified by chromatography across a nickle-nitrilotriacetic acid-agarose column under denaturing conditions. the purified his-tagged wild-type protein could be functionally reconstituted in the presence of detergent and sterol and ...19968662769
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
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
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
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
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
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
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 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
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
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
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
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
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
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
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
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
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
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
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
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
phosphate transporters from the higher plant arabidopsis thaliana.two cdnas (atpt1 and atpt2) encoding plant phosphate transporters have been isolated from a library prepared with mrna extracted from phosphate-starved arabidopsis thaliana roots, the encoded polypeptides are 78% identical to each other and show high degree of amino acid sequence similarity with high-affinity phosphate transporters of saccharomyces cerevisiae, neurospora crassa, and the mycorrhizal fungus glomus versiforme. the atpt1 and atpt2 polypeptides are integral membrane proteins predicte ...19968927627
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
isolation and characterization of a laccase gene from podospora anserina.the genome of the filamentous ascomycete podospora anserina contains at least four non-adjacent regions that are homologous to the laccase gene of neurospora crassa. one of these regions contains a gene (lac2) encoding a protein that displays 62% identity with the n. crassa laccase. in shaken cultures, lac2 mrna is present at low basal levels throughout the growth phase but increases at least 20-fold at the beginning of the autolytic phase and decreases again thereafter. addition of aromatic xen ...19968914515
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