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biogenesis and function of the yeast plasma-membrane h(+)-atpase.one of the most abundant proteins in the yeast plasma membrane is the p-type h(+)-atpase that pumps protons out of the cell, supplying the driving force for a wide array of h(+)-dependent cotransporters. the atpase is a 100 kda polypeptide, anchored in the lipid bilayer by 10 transmembrane alpha-helices. it is structurally and functionally related to the p-type na(+),k(+)-, h(+),k(+)- and ca(2+)-atpases of animal cells and the h(+)-atpases of plant cells, and it shares with them a characteristic ...200010600684
analysis of the membrane domain of the gastric h(+)/k(+)-atpase.a structure of the catalytic or alpha subunit of the h(+)/k(+)-atpase, with ten transmembrane segments, and of the beta subunit, with a single such segment, was established using a combination of tryptic cleavage and peptide sequencing and in vitro translation. sites at which covalent ligands bind to external surfaces were also defined by cleavage, separation and sequencing. cys813 was found to be the common covalent binding site for all the substituted pyridyl methylsulfinyl benzimidazoles. the ...200010600685
posttranscriptional gene silencing in neurospora by a recq dna helicase.the phenomenon of posttranscriptional gene silencing (ptgs), which occurs when a transgene is introduced into a cell, is poorly understood. here, the qde-3 gene, which is required for the activation and maintenance of gene silencing in the fungus neurospora crassa, was isolated. sequence analysis revealed that the qde-3 gene belongs to the recq dna helicase family. the qde3 protein may function in the dna-dna interaction between introduced transgenes or with an endogenous gene required for gene- ...199910600745
a highly conserved mechanism of regulated ribosome stalling mediated by fungal arginine attenuator peptides that appears independent of the charging status of arginyl-trnas.the arg attenuator peptide (aap) is an evolutionarily conserved peptide involved in arg-specific negative translational control. it is encoded as an upstream open reading frame (uorf) in fungal mrnas specifying the small subunit of arg-specific carbamoyl phosphate synthetase. we examined the functions of the saccharomyces cerevisiae cpa1 and neurospora crassa arg-2 aaps using translation extracts from s. cerevisiae, n. crassa, and wheat germ. synthetic rna containing aap and firefly luciferase ( ...199910608810
structural and functional definition of the human chitinase chitin-binding domain.mammalian chitinase, a chitinolytic enzyme expressed by macrophages, has been detected in atherosclerotic plaques and is elevated in blood and tissues of guinea pigs infected with aspergillus. its normal physiological function is unknown. to understand how the enzyme interacts with its substrate, we have characterized the chitin-binding domain. the c-terminal 49 amino acids make up the minimal sequence required for chitin binding activity. the absence of this domain does not affect the ability o ...200010617646
use of atp6 in fungal phylogenetics: an example from the boletales.complete nucleotide sequences have been determined for atp6 from suillus luteus and cox3 from suillus sinuspaulianus (boletales, hymenomycetes, basidiomycota), which code for atpase subunit 6 and cytochrome oxidase subunit 3, respectively. these sequences were used to design pcr primers for the amplification of partial atp6 and cox3 sequences from other members of the boletales and outgroup taxa. in atp6 and cox3 from russula rosacea, one of the outgroup taxa, we observed a number of in-frame tg ...199910620406
cloning and characterization of a gene (arpa) from aspergillus oryzae encoding an actin-related protein required for normal nuclear distribution and morphology of conidiophores.we have isolated the arpa gene from aspergillus oryzae as a homologue of the neurospora crassa ro-4 gene. in n. crassa, mutations in the ro-4 gene, which encodes a major component of the dynactin complex arp1, causes curling of hyphae and abnormalities in nuclear distribution. the arpa gene contains two introns and encodes a polypeptide of 381 amino acids, with a 78% sequence identity to the n. crassa arp1. overexpression of the arpa gene causes a defect in nuclear migration into elongating hyph ...199910628858
in vitro antifungal evaluation and studies on the mode of action of xanthoxyline derivatives.this study describes the fungistatic effect of xanthoxyline (cas 90-24-4) and its derivatives against a panel of yeasts, filamentous fungi and dermatophytes, by using the agar dilution method. results indicated that simple structural modifications led to more potent derivatives, especially in relation with dermatophytes. the most active compound tested (10), which is a benzenesulphonyl derivative, was 12-fold more potent than xanthoxyline itself against trichophyton rubrum. the evaluation of the ...199910635452
a fission yeast kinesin affects golgi membrane recycling.we report here an in vivo study of kinesin heavy chain (khc) functions in yeast. we have identified in schizosaccharomyces pombe a kinesin motor gene, klp3(+), which has the highest homology to the neurospora crassa khc. using indirect immunofluorescence, ha epitope-tagged klp3 protein is cytoplasmic and appears as one to a few distinct patches that are coincident with microtubules. the klp3 null allele is viable. in klp3 deleted cells, er, golgi and mitochondrial distribution appear normal. mit ...200010641037
suppressed recombination and a pairing anomaly on the mating-type chromosome of neurospora tetrasperma.neurospora crassa and related heterothallic ascomycetes produce eight homokaryotic self-sterile ascospores per ascus. in contrast, asci of n. tetrasperma contain four self-fertile ascospores each with nuclei of both mating types (mata and mata). the self-fertile ascospores of n. tetrasperma result from first-division segregation of mating type and nuclear spindle overlap at the second meiotic division and at a subsequent mitotic division. recently, merino et al. presented population-genetic evid ...200010655216
specific binding sites for an antifungal plant defensin from dahlia (dahlia merckii) on fungal cells are required for antifungal activity.dm-amp1, an antifungal plant defensin from seeds of dahlia (dahlia merckii), was radioactively labeled with t-butoxycarbonyl-[35s]-l-methionine n-hydroxy-succinimi-dylester. this procedure yielded a 35s-labeled peptide with unaltered antifungal activity. [35s]dm-amp1 was used to assess binding on living cells of the filamentous fungus neurospora crassa and the unicellular fungus saccharomyces cerevisiae. binding of [35s]dm-amp1 to fungal cells was saturable and could be competed for by preincuba ...200010656585
comparative analysis of ca2+ and h+ flux magnitude and location along growing hyphae of saprolegnia ferax and neurospora crassa.calcium and proton ion fluxes were mapped at the growing apices of two hyphal organisms, the oomycete saprolegnia ferax and the ascomycete neurospora crassa and pseudohyphal saccharomyces cerevisiae using self-referencing ion-selective probes. s. ferax exhibited well-defined transport zones absent in n. crassa. ca2+ fluxes were located within 8 microm of the growing hyphal tip; the net ca2+ flux was either inward (75% of all experiments) or outward. the inward component of the net flux was inhib ...199910669108
the frost gene of neurospora crassa is a homolog of yeast cdc1 and affects hyphal branching via manganese homeostasis.the neurospora crassa mutant frost has a hyperbranching phenotype that can be corrected by adding ca(2+), suggesting that characterization of this gene might clarify the mechanism of ca(2+)-dependent tip growth. the wild-type allele was cloned by sib selection using protoplasts from arthroconidia. rflp analysis revealed that the cloned dna fragment mapped to the fr locus. the nucleotide sequence of genomic and cdna was determined. the deduced amino acid sequence showed homology to the saccharomy ...199910669587
kalilo plasmids are a family of four distinct members with individual global distributions across species.kalilo is a linear 9-kb plasmid, isolated originally from hawaiian strains of the heterothallic fungus neurospora intermedia. its properties include terminal inverted repeats, two orfs coding for a presumptive dna and an rna polymerase, and the ability to cause senescence in its original host and in the closely related species neurospora crassa. we have examined natural isolates alleged to contain plasmids homologous to kalilo. most of these isolates do in fact contain plasmids with so close an ...200010672443
a non-stop antisense reading frame in the grp78 gene of neurospora crassa is homologous to the achlya klebsiana nad-gdh gene but is not being transcribed.a long non-stop reading frame exists on the antisense strand of the grp78 gene (cdna and genomic dna) of neurospora crassa. computer analysis revealed a strong similarity of the putative antisense protein to the 10th exon of the nad-dependent glutamate dehydrogenase gene (nad-gdh) of achlya klebsiana, which is itself located on the complementary strand of a transcribed hsc70 gene homologue. in neurospora, no grp78 antisense mrna was detected by northern blot and reverse transcription-coupled pol ...200010675602
molecular cloning of the calcium and sodium atpases in neurospora crassa.using pcr, reverse transcription-pcr (rt-pcr) and colony hybridization in a genomic library, we isolated six genes which encode type ii p-type atpases in neurospora crassa. the six full-length cdnas were cloned in a yeast expression vector and transformed into saccharomyces cerevisiae null ca2+- or na+-atpase mutants. three cdnas suppressed the defect of the ca2+ mutant and two of these protected from mn2+ toxicity. one cdna suppressed the defect of the na+ mutant and two cdnas were not function ...200010712689
isolation of trypanosoma brucei cyc2 and cyc3 cyclin genes by rescue of a yeast g(1) cyclin mutant. functional characterization of cyc2.two trypanosoma brucei cyclin genes, cyc2 and cyc3, have been isolated by rescue of the saccharomyces cerevisiae mutant dl1, which is deficient in cln g(1) cyclin function. cyc2 encodes a 24-kda protein that has sequence identity to the neurospora crassa preg1 and the s. cerevisiae pho80 cyclin. cyc3 has the most sequence identity to mitotic b-type cyclins from a variety of organisms. both cyc2 and cyc3 are single-copy genes and expressed in all life cycle stages of the parasite. to determine if ...200010722661
molecular cloning and functional analysis of apoxin i, a snake venom-derived apoptosis-inducing factor with l-amino acid oxidase activity.we previously purified apoxin i, an apoptosis-inducing factor with l-amino acid oxidase (lao) activity, from western diamondback rattlesnake venom. to determine the primary structure of apoxin i, we cloned its cdna. the amino acid sequence showed that apoxin i has an fad binding domain and shares homology with l-amino acid oxidase (lao) from neurospora crassa, human monoamine oxidase b, and mouse interleukin 4-induced f1g1 protein. the full-length apoxin i has an n-terminal signal sequence that ...200010727211
cyclophilin-promoted folding of mouse dihydrofolate reductase does not include the slow conversion of the late-folding intermediate to the active enzyme.cyclophilins accelerate slow protein folding reactions in vitro by catalyzing the cis/trans isomerization of peptidyl-prolyl bonds. cyclophilins were reported to be involved in a variety of cellular functions, including the promotion of protein folding by use of the substrate mouse dihydrofolate reductase (dhfr). the interaction of cyclophilin with dhfr has only been studied under limited conditions so far, not taking into account that native dhfr exists in equilibrium with a non-native late-fol ...200010731431
construction and expression of functional multi-domain polypeptides in escherichia coli: expression of the neurospora crassa metallothionein gene.a system for the construction of polymeric peptides in escherichia coli was utilized to prepare a library of plasmids coding for tandem repeats of the neurospora crassa metallothionein gene. selected oligomeric metallothionein clones were expressed and targeted to the periplasm as a fusion with the maltose-binding protein. bacterial cells harbouring the expressed oligopeptides were characterized for their ability to bind 109cd2+. the metal-binding ability was enhanced for all the oligomeric cons ...200010736021
a rapid technique for screening of lovastatin-producing strains of aspergillus terreus by agar plug and neurospora crassa bioassay.the success of strain improvement programme depends on the number of isolates that can be screened after mutagenic treatment. a technique to rapidly screen large number of high-yielding isolates was developed. the 'agar plug' method that utilizes the anti-fungal property of lovastatin to produce a zone of inhibition against neurospora crassa was not only economical but also less labour-intensive. we were able to isolate a high-yielding strain, the productivity of which increased by 138% as compa ...200010739348
characterization and expression of a neurospora crassa ribosomal protein gene, crp-7.we have isolated and characterized a neurospora crassa cytoplasmic ribosomal protein gene, named crp-7, which is found upstream of the photolyase gene. the deduced amino-acid sequence of this gene is highly homologous to the ys25 ribosomal protein of saccharomyces cerevisiae. the crp-7 orf consists of two exons which are separated by a short intron. the deduced polypeptide contains 87 amino acid residues and has a calculated molecular weight of 9.7 kda. rflp mapping showed that the crp-7 gene is ...200010743568
protein transport into mitochondria.mitochondria are made up of two membrane systems that subdivide this organelle into two aqueous subcompartments: the matrix, which is enclosed by the inner membrane, and the intermembrane space, which is located between the inner and the outer membrane. protein import into mitochondria is a complex reaction, as every protein has to be routed to its specific destination within the organelle. in the past few years, studies with mitochondria of neurospora crassa and saccharomyces cerevisiae have le ...200010744987
interaction of the neurospora crassa heat shock factor with the heat shock element during heat shock and different developmental stages.the interaction of the heat shock factor (hsf) with the heat shock element (hse) was determined by a non-radioactive electrophoretic mobility shift assay, in order to analyze hsf regulation in neurospora crassa. hsf binds to hse under normal, non-stress conditions and is thus constitutively trimerized. upon heat shock, the hsf-hse complex shows a retarded mobility. this was also observed in saccharomyces cerevisiae, where this mobility shift was shown to be due to hsf phosphorylation [sorger and ...200010754257
molecular analysis of the aureobasidium pullulans ura3 gene encoding orotidine-5'-phosphate decarboxylase and isolation of mutants defective in this gene.orotidine-5'-phosphate decarboxylase (omp decarboxylase) catalyses the final step in the pyrimidine biosynthesis, the conversion of orotidine-5'-phosphate (omp) to uridine-5'-phosphate. the ura3 gene of aureobasidium pullulans. encoding omp decarboxylase, was isolated from an aureobasidium genomic library constructed in the plasmid pblueskriptsk-. the ura3 gene of a. pullulans has an open reading frame of 271 amino acid residues. analysis of the sequence revealed the presence of two introns. in ...200010772469
tagging of a nitrogen pathway-specific regulator gene in tolypocladium inflatum by the transposon restless.restless is an endogenous hat transposon found in the cyclosporin-producing fungus tolypocladium inflatum. this element is present in about 15 copies in a particular strain (atcc34921) which was used for successful gene tagging. we have isolated a t. inflatum mutant with a defect in nitrogen metabolism. this mutant carries a copy of the restless element in a gene encoding a c6 zinc-finger protein. the deduced amino acid sequence of the gene product shows a significant similarity to the nit4 prot ...200010778749
understanding circadian rhythmicity in neurospora crassa: from behavior to genes and back again.circadian clocks have been described in organisms ranging in complexity from unicells to mammals, in which they function to control daily rhythms in cellular activities and behavior. the significance of a detailed understanding of the clock can be appreciated by its ubiquity and its established involvement in human physiology, including endocrine function, sleep/wake cycles, psychiatric illness, and drug tolerances and effectiveness. because the clock in all organisms is assembled within the cel ...200010779395
large-scale comparison of fungal sequence information: mechanisms of innovation in neurospora crassa and gene loss in saccharomyces cerevisiae.we report a large-scale comparison of sequence data from the filamentous fungus neurospora crassa with the complete genome sequence of saccharomyces cerevisiae. n. crassa is considerably more morphologically and developmentally complex than s. cerevisiae. we found that n. crassa has a much higher proportion of "orphan" genes than s. cerevisiae, suggesting that its morphological complexity reflects the acquisition or maintenance of novel genes, consistent with its larger genome. our results also ...200010779483
a new self-assembled peroxisomal vesicle required for efficient resealing of the plasma membrane.the woronin body is a membrane-bound organelle that has been observed in over 50 species of filamentous fungi. however, neither the composition nor the precise function of the woronin body has yet been determined. here we purify the woronin body from neurospora crassa and isolate hex1, a new protein containing a consensus sequence known as peroxisome-targeting signal-1 (pts1). we show that hex1 is localized to the matrix of the woronin body by immunoelectron microscopy, and that a green fluoresc ...200010783241
the bacillus subtilis cysp gene encodes a novel sulphate permease related to the inorganic phosphate transporter (pit) family.sulphate permeases in the plasma membrane are responsible for uptake of environmental sulphate used in the sulphate assimilation pathway in bacteria and plants. here it is reported that the orf designated cysp, located on the bacillus subtilis chromosome between cysh and five putative genes involved in sulphate assimilation, encodes a sulphate permease. cysp is able to complement escherichia coli cysteine auxotrophs with mutations affecting either the membrane or periplasmic components of the su ...200010784039
a nuclease from neurospora crassa specific for d(a-t) rich regions in double stranded dna.a nuclease from n. crassa has been prepared to the hydroxylapatite stage of purification described by rabin and fraser (1). it degrades single stranded dna in an essentially exonucleolytic process. it does not give any appreciable acid soluble material with double stranded dna as substrate. this shows its high degree of specificity towards single stranded dna. an extra activity which makes double strand breaks in the at rich regions of dna is described. a limited number of breaks are observed wi ...197410793677
isolation of repeated and self-complementary sequences from e. coli dna.we have used the single-strand specific nuclease from neurospora crassa and chromatography on methylated albumin-kieselguhr to purify and characterize repeated and self-complementary sequences from escherichia coli dna. approximately 0.5% of the genome renatures spontaneously at zero time and another 2% renatures somewhat more rapidly than the total dna. the early renaturing dna has a base composition and a tm similar to the total dna and contains on the average 100 base pairs; the self-compleme ...197410793709
methylation of the foreign transposon restless in vegetative mycelia of neurospora crassa.methylation of foreign and/or repeated sequences in the filamentous fungus neurospora crassa is believed to be directed against invading transposable elements. to test this hypothesis, the fate of a transposon in n. crassa was investigated. vectors were constructed which carried the transposon restless, an active class-ii element isolated from the fungus tolypocladium inflatum. these vectors were introduced into n. crassa strains by protoplast transformation. two strategies were employed: (1) ec ...200010794177
presence of a pertussis toxin-sensitive g protein alpha subunit in sporothrix schenckii.as an initial step in the study of the role of g proteins in signal transduction in sporothrix schenckii, we identified a galphai subunit using different experimental approaches. western blots of fungal membrane preparations using anti-galphacommon and anti-galphai1-galphai2 antibodies identified a band of approximately 41 kda. pertussis toxin-catalyzed adenosine diphosphate (adp)-ribosylation of these membrane fractions confirmed the presence of a protein substrate of 41 kda. a 357 bp polymeras ...200010817227
evolutionarily conserved features of the arginine attenuator peptide provide the necessary requirements for its function in translational regulation.neurospora crassa arg-2 mrna contains an evolutionarily conserved upstream open reading frame (uorf) encoding the arg attenuator peptide (aap) that confers negative translational regulation in response to arg. we examined the regulatory role of the aap and the rna encoding it using an n. crassa cell-free translation system. aaps encoded by uorfs in four fungal mrnas each conferred negative regulation in response to arg by causing ribosome stalling at the uorf termination codon. deleting the aap ...200010818103
immunolocalization of ca ii and h+ v-atpase in epithelial cells of the mouse and rat epididymis.acidification of the epididymal lumen has been suggested to play an important role in sperm functions; however, the cell types, pumps, and mechanisms involved have not been fully addressed. in this study, carbonic anhydrase ii (ca ii) and a 67-kd subunit of neurospora crassa vacuolar proton adenosinetriphosphatase (h+ v-atpase) pump were immunolocalized using light microscopy and electron microscopy (em) in the epididymis of rats and mice. in both animals, narrow cells, identified in the initial ...200010819445
confocal microscopy of fm4-64 as a tool for analysing endocytosis and vesicle trafficking in living fungal hyphae.confocal microscopy of amphiphilic styryl dyes has been used to investigate endocytosis and vesicle trafficking in living fungal hyphae. hyphae were treated with fm4-64, fm1-43 or tma-dph, three of the most commonly used membrane-selective dyes reported as markers of endocytosis. all three dyes were rapidly internalized within hyphae. fm4-64 was found best for imaging the dynamic changes in size, morphology and position of the apical vesicle cluster within growing hyphal tips because of its stai ...200010849201
cloning and expression of the s-adenosylmethionine decarboxylase gene of neurospora crassa and processing of its product.s-adenosylmethionine decarboxylase (adometdc) catalyzes the formation of decarboxylated adometdc, a precursor of the polyamines spermidine and spermine. the enzyme is derived from a proenzyme by autocatalytic cleavage. we report the cloning and regulation of the gene for adometdc in neurospora crassa, spe-2, and the effect of putrescine on enzyme maturation and activity. the gene was cloned from a genomic library by complementation of a spe-2 mutant. like other adometdcs, that of neurospora is d ...200010852489
tagging of genes involved in multidrug resistance in aspergillus nidulans.we have used a plasmid containing the neurospora crassa pyr4 gene to transform an aspergillus nidulans pyrg89 mutant strain in the presence of bam-hi, and isolated multidrug-sensitive mutants among the transformants. using this approach, we hoped to identify genes whose products are important for drug resistance by analyzing gene disruptions that alter the drug sensitivity of the cell. about 1300 transformants isolated following transformation were screened for sensitivity to drugs or various st ...200010852493
differential complementation of a neurospora crassa galpha(i) mutation using mammalian galpha protein genes.heterotrimeric (alphabetagamma) g proteins interact with sensory receptors to transduce signals to downstream effectors in eukaryotes. we previously reported that gna-1 from neurospora crassa is a microbial member of the galphai family found in higher organisms. deletion of gna-1 leads to female sterility, slower growth rates on normal and hyperosmotic solid medium, and increased resistance to heat and oxidative stress. in this study we compare mammalian genes for proteins of the galphai sub-fam ...200010852494
import of a cytosolic protein into lysosomes by chaperone-mediated autophagy depends on its folding state.we have analyzed the folding state of cytosolic proteins imported in vitro into lysosomes, using an approach originally developed by eilers and schatz, (eilers, m., and schatz, g. (1986) nature 322, 228-232) to investigate protein import into mitochondria. the susceptibility toward proteases of mouse dihydrofolate reductase (dhfr), synthesized in a coupled transcription-translation system with rabbit reticulocytes, decreased in the presence of its substrate analogue, methotrexate. this analogue ...200010862611
two approaches to isolate cytoplasmic dynein atpase from neurospora crassa.cytoplasmic dynein is a force-producing enzyme that, in association with dynactin, conducts minus-end directed transport of various organelles along microtubules. biochemical analyses of cytoplasmic dynein and dynactin have been conducted primarily in vertebrate systems, whereas genetic analyses have been explored mainly in yeast and the filamentous fungi. to provide a complementary biochemical approach for the study of fungal dynein, we isolated/partially purified cytoplasmic dynein atpase from ...200010863006
histidine kinase, two-component signal transduction proteins of candida albicans and the pathogenesis of candidosis.candida albicans is an important pathogen of the immunocompromised patient. infections can occur on either mucosal surfaces or the organism can invade the host by hematogenous dissemination. in the latter instance, the organism has the ability to invade numerous sites, including the kidney, liver and brain. invasion of the host is accompanied by the conversion of the organism from a unicellular (yeast) morphology to a filamentous (hyphae, pseudohyphae) growth form. the morphogenetic change which ...199910865904
blue-light regulation of phytoene dehydrogenase (carb) gene expression in mucor circinelloides.the carb gene, encoding the phytoene dehydrogenase of mucor circinelloides, was isolated by heterologous hybridisation with a probe derived from the corresponding gene of phycomyces blakesleeanus. the cdna and genomic copies complemented phytoene dehydrogenase defects in escherichia coli and in carb mutants of m. circinelloides, respectively. fluence-response curves for transcript accumulation were constructed after different blue-light pulses. the level of carb mrna accumulation reached values ...200010872226
nucleotide sequence and intron structure of the apocytochrome b gene of neurospora crassa mitochondria.the sequence of the apocytochrome b (cob) gene of neurospora crassa has been determined. the structural gene is interrupted by two intervening sequences of approximately 1260 bp each. the polypeptide encoded by the exons shows extensive homology with the cob proteins of aspergillus nidulans and saccharomyces cerevisiae (79% and 60%, respectively). the two introns are, however, located at sites different from those of introns in the cob genes of a. nidulans and s. cerevisiae (which contain highly ...198310872314
vegetative incompatibility in the het-6 region of neurospora crassa is mediated by two linked genes.non-self-recognition during asexual growth of neurospora crassa involves restriction of heterokaryon formation via genetic differences at 11 het loci, including mating type. the het-6 locus maps to a 250-kbp region of lgiil. we used restriction fragment length polymorphisms in progeny with crossovers in the het-6 region and a dna transformation assay to identify two genes in a 25-kbp region that have vegetative incompatibility activity. the predicted product of one of these genes, which we desig ...200010880472
a tip-high gradient of a putative plasma membrane snare approximates the exocytotic gradient in hyphal apices of the fungus neurospora crassa.antibodies to the saccharomyces cereviseae plasma membrane t-snare sso2p identify a putative 39-kda homologue in neurospora crassa. the 39-kda protein is enriched in plasma membrane (pm) and occurred with other membranes. it is extractable by detergent, but not chaotropic or alkali agents, suggesting membrane insertion. immunoprecipitation with anti-sso2p coprecipitated a approximately 100-kda, mg(+)-atp-sensitive band with the 39-kda protein, suggesting a ternary snare complex. affinity-purifie ...200010882535
interconnected feedback loops in the neurospora circadian system.in neurospora crassa, white collar 1 (wc-1), a transcriptional activator and positive clock element, is rhythmically expressed from a nonrhythmic steady-state pool of wc-1 transcript, consistent with posttranscriptional regulation of rhythmicity. mutations in frq influence both the level and periodicity of wc-1 expression, and driven frq expression not only depresses its own endogenous levels, but positively regulates wc-1 synthesis with a lag of about 8 hours, a delay similar to that seen in th ...200010884222
biosynthesis of pyridoxine: origin of the nitrogen atom of pyridoxine in microorganisms.the amide nitrogen atom of glutamine was incorporated into pyridoxine in four eukaryotes, emericella nidulans, mucor racemosus, neurospora crassa and saccharomyces cerevisiae, and two prokaryotes, staphylococcus aureus and bacillus subtilis, but not in the following prokaryotes, pseudomonas putida, enterobacter aerogenes and escherichia coli. on the other hand, the nitrogen atom of glutamate was incorporated into pyridoxine in p. putida, e. aerogenes and e. coli, but not in s. aureus and b. subt ...200010885790
the tryptophan synthase-encoding trpb gene of aspergillus nidulans is regulated by the cross-pathway control system.the tryptophan synthase-encoding gene, trpb, of aspergillus nidulans was cloned and characterized. it was mapped to chromosome i, between the gene meda, which is required for sexual and asexual development, and an orf encoding a protein with significant similarity to subunit b of vacuolar atp synthases. the 5' untranslated region was found to be at least 142 nucleotides (nt) long, the poly(a) addition site was localized at position + 216 relative to the stop codon by sequencing of several indepe ...200010905354
cloning and characterisation of a chitin synthase gene cdna from the cultivated mushroom agaricus bisporus and its expression during morphogenesis.full-length cdna of a chitin synthase gene (chs1) was cloned from agaricus bisporus by screening a cdna library with a pcr amplified fragment of the chitin synthase gene. the chs1 contains an open reading frame of 2727 bp encoding a polypeptide of 909 amino acids and deduced molecular mass 102.3 kda and pi 8.23. the central region of chs1 showed strong homology to other fungal chitin synthase genes with seven conserved domains. it belongs to the chitin synthase class iii, analogous to chsb from ...200010913868
a group of expressed cdna sequences from the wheat fungal leaf blotch pathogen, mycosphaerella graminicola (septoria tritici).a group of expressed sequence tags (ests) from the wheat fungal pathogen mycosphaerella graminicola utilizing ammonium as a nitrogen source has been analyzed. single pass sequences of complementary dnas from 986 clones were determined. contig analysis and sequence comparisons allowed 704 unique ests (unigenes) to be identified, of which 148 appeared as multiple copies. searches of the nrdb95 protein database at embl using the blast2x algorithm revealed 407 (57.8%) sequences that generated high t ...200010919380
function of tyrosyl-trna synthetase in splicing group i introns: an induced-fit model for binding to the p4-p6 domain based on analysis of mutations at the junction of the p4-p6 stacked helices.we used an escherichia coli genetic assay based on the phage t4 td intron to test the ability of the neurospora crassa mitochondrial tyrosyl-trna synthetase (cyt-18 protein) to suppress mutations that cause structural defects around its binding site in the p4-p6 domain of the group i intron catalytic core. we analyzed all possible combinations of nucleotides at either p4 bp-1 or p6 bp-1, which together form the junction of the p4-p6 stacked helices, and looked for synergistic effects in double m ...200010926509
the lis1-related nudf protein of aspergillus nidulans interacts with the coiled-coil domain of the nude/ro11 protein.the nudf gene of the filamentous fungus aspergillus nidulans acts in the cytoplasmic dynein/dynactin pathway and is required for distribution of nuclei. nudf protein, the product of the nudf gene, displays 42% sequence identity with the human protein lis1 required for neuronal migration. haploinsufficiency of the lis1 gene causes a malformation of the human brain known as lissencephaly. we screened for multicopy suppressors of a mutation in the nudf gene. the product of the nude gene isolated in ...200010931877
characterization of hch, the podospora anserina homolog of the het-c heterokaryon incompatibility gene of neurospora crassa.the het-c locus controls heterokaryon formation in neurospora crassa. it is subject to balancing selection operating to maintain polymorphism at that locus in natural populations. we have isolated hch, the het-c homolog from the related species podospora anserina (hch for het-c homolog), in order to determine if this gene also functions as a het gene in that species. the het-c and hch sequences are highly similar but differ in the region defining allele specificity in n. crassa het-c. analysis o ...200010953880
the degenerate dna transposon pat and repeat-induced point mutation (rip) in podospora anserina.a degenerate dna transposon, pat, was identified in the genomes of various wild-type strains of the filamentous fungus podospora anserina. in these strains, the number (approximately 20-25 copies per genome) and location of pat sequences appear to be conserved. two copies of pat, one complete and one partial, were cloned and characterized. the sequence of the complete element is 1856 bp long and contains imperfect inverted terminal repeats (itrs) of 53 bp. the target site duplication comprises t ...200010954093
plasma membrane-adjacent actin filaments, but not microtubules, are essential for both polarization and hyphal tip morphogenesis in saprolegnia ferax and neurospora crassa.the organization and roles of f-actin and microtubules in the maintenance and initiation of hyphal tip growth have been analyzed in saprolegnia ferax and neurospora crassa. in hyphae of both species, the apex is depleted of microtubules relative to subapical regions and near-normal morphogenesis occurs in concentrations of nocodazole or mbc which remove microtubules, slow growth, and disrupt nuclear positioning. in contrast, each species contains characteristic tip-high arrays of plasma membrane ...200010955907
molecular genetics of heterokaryon incompatibility in filamentous ascomycetes.filamentous fungi spontaneously undergo vegetative cell fusion events within but also between individuals. these cell fusions (anastomoses) lead to cytoplasmic mixing and to the formation of vegetative heterokaryons (i.e., cells containing different nuclear types). the viability of these heterokaryons is genetically controlled by specific loci termed het loci (for heterokaryon incompatibility). heterokaryotic cells formed between individuals of unlike het genotypes undergo a characteristic cell ...200010974123
role of mmm1 in maintaining mitochondrial morphology in neurospora crassa.mmm1p is a protein required for maintenance of mitochondrial morphology in budding yeast. it was proposed that it is required to mediate the interaction of the mitochondrial outer membrane with the actin cytoskeleton. we report the cloning and characterization of mmm1 of the filamentous fungus neurospora crassa, an organism that uses microtubules for mitochondrial transport. mutation of the mmm-1 gene leads to a temperature-sensitive slow growth phenotype and female sterility. mutant cells harbo ...200010982393
comparison of chitin content in the apical and distal parts of fungal hyphae in basidiobolus ranarum, neurospora crassa and coprinus sterquilinus.primary cell wall is synthesized in the growth zone of hyphal apex in fungi and rigidified during maturation along the newly formed hypha. cross-linking of cell-wall components and self-assembly of individual polysaccharide chains into microfibrils are supposed to be involved in the rigidification process. we determined the relative chitin content in the cell wall of hyphal tips and distal walls of three fungal species and demonstrated a general increase in relative chitin content in mature cell ...199910983235
asd4, a new gata factor of neurospora crassa, displays sequence-specific dna binding and functions in ascus and ascospore development.a new gene encoding a novel gata factor, asd4, of neurospora crassa was isolated and demonstrated to possess one intron and to specify an open reading frame encoding a protein with 427 amino acid residues. the asd4 protein contains a single gata-type zinc finger and a putative coiled-coil domain. unlike related proteins, dal80 in yeast and nreb in penicillium, asd4 does not appear to be involved in regulation of nitrogen metabolism. an asd-4 null mutant obtained by the rip procedure did not show ...200010998244
regulation of conidiation and adenylyl cyclase levels by the galpha protein gna-3 in neurospora crassa.we have identified a new gene encoding the g protein alpha subunit, gna-3, from the filamentous fungus neurospora crassa. the predicted amino acid sequence of gna-3 is most similar to the galpha proteins mod-d, maga, and cpg-2 from the saprophytic fungus podospora anserina and the pathogenic fungi magnaporthe grisea and cryphonectria parasitica, respectively. deletion of gna-3 leads to shorter aerial hyphae and premature, dense conidiation during growth on solid medium or in standing liquid cult ...200011003665
characterization of two novel redox groups in the respiratory nadh:ubiquinone oxidoreductase (complex i).the proton-pumping nadh:ubiquinone oxidoreductase is the first of the respiratory chain complexes in many bacteria and mitochondria of most eukaryotes. the bacterial complex consists of 14 different subunits. seven peripheral subunits bear all known redox groups of complex i, namely one fmn and five epr-detectable iron-sulfur (fes) clusters. the remaining seven subunits are hydrophobic proteins predicted to fold into 54 alpha-helices across the membrane. little is known about their function, but ...200011004444
characterization of mrad18sc, a mouse homolog of the yeast postreplication repair gene rad18.the rad18 gene of the yeast saccharomyces cerevisiae encodes a protein with ssdna binding activity that interacts with the ubiquitin-conjugating enzyme rad6 and plays an important role in postreplication repair. we identified and characterized the putative mouse homolog of rad18, designated mrad18sc. the mrad18sc open reading frame encodes a 509-amino-acid polypeptide that is strongly conserved in size and sequence between yeast and mammals, with specific conservation of the ring-zinc-finger and ...200011013078
characterization of the neurospora crassa mus-25 mutant: the gene encodes a protein which is homologous to the saccharomyces cerevisiae rad54 protein.characterization of the neurospora crassa mus-25 mutant suggests that it is defective in recombination repair and belongs to the uvs-6 epistasis group. it shows a high sensitivity to the alkylating agents methyl methanesulfonate (mms) and n-methyl-n'-nitro-n-nitrosoguanidine (mnng), but not to uv radiation. it is barren (i.e. does not produce ascospores) in homozygous crosses. the frequency of mms-induced mutations at the ad-3 loci is approximately three times higher than in the wild type. the r ...200011016845
a single five-step desaturase is involved in the carotenoid biosynthesis pathway to beta-carotene and torulene in neurospora crassa.phytoene desaturase al-1 from neurospora crassa was expressed in escherichia coli and an active enzyme was isolated which catalyzed the stepwise introduction of up to five double bonds into the substrate phytoene. the major reaction products were 3, 4-didehydrolycopene and lycopene. several of the desaturation intermediates, zeta-carotene, neurosporene, and lycopene, were also accepted as a substrate by al-1. in contrast to the structurally related bacterial enzymes, the cofactor involved in the ...200011017770
antisense expression of the calmodulin gene from colletotrichum trifolii impairs prepenetration development(1).calmodulin is a ubiquitous highly conserved calcium binding protein involved in cell signalling. previous studies in our laboratory suggested a role for calmodulin in prepenetration morphogenesis in colletotrichum trifolii, the causal agent of alfalfa anthracnose. in this report, we describe the cloning, sequencing and partial characterization of the calmodulin gene from c. trifolii. the gene is present as a single copy in the genome of c. trifolii and its predicted amino acid sequence shows con ...200011024266
a secondary beta deuterium kinetic isotope effect in the chorismate synthase reaction.chorismate synthase (ec 4.6.1.4) is the shikimate pathway enzyme that catalyzes the conversion of 5-enolpyruvylshikimate 3-phosphate (epsp) to chorismate. the enzyme reaction is unusual because it involves a trans-1,4 elimination of the c-3 phosphate and the c-6 pror hydrogen and it has an absolute requirement for reduced flavin. several mechanisms have been proposed to account for the cofactor requirement and stereochemistry of the reaction, including a radical mechanism. this paper describes t ...200011034781
functional analysis of the leader peptide of the yeast gene cpa1 and heterologous regulation by other fungal peptides.the 25-amino-acid leader peptide present at the 5' end of yeast cpa1 mrna is responsible for the translational repression of that gene by arginine. we show here that the active domain of the yeast peptide is highly specific and extends over amino acids 6-23. the region between amino acids 6-21 is well conserved between similar peptides present upstream of cpa1-homologous genes in other fungi. the neurospora crassa arg-2 peptide represses the expression of cpa1, whereas the peptide from aspergill ...200011057443
yeti--a degenerate gypsy-like ltr retrotransposon in the filamentous ascomycete podospora anserina.in the filamentous ascomycete podospora anserina a 6,935-bp retrotransposon, yeti, has been identified and characterized. it is flanked by a 5-bp target site duplication and contains long terminal repeats (ltrs) 354 bp in length. the ltrs show a high degree of identity to the previously reported repetitive element repa, a sequence suggested to represent a solo-ltr element of an unknown transposon. in the investigated podospora strains, the number of complete yeti copies is significantly lower th ...200011057446
gene cloning and molecular characterization of lysine decarboxylase from selenomonas ruminantium delineate its evolutionary relationship to ornithine decarboxylases from eukaryotes.lysine decarboxylase (ldc; ec 4.1.1.18) from selenomonas ruminantium comprises two identical monomeric subunits of 43 kda and has decarboxylating activities toward both l-lysine and l-ornithine with similar k(m) and v(max) values (y. takatsuka, m. onoda, t. sugiyama, k. muramoto, t. tomita, and y. kamio, biosci. biotechnol. biochem. 62:1063-1069, 1999). here, the ldc-encoding gene (ldc) of this bacterium was cloned and characterized. dna sequencing analysis revealed that the amino acid sequence ...200011073919
a neurospora double-strand-break repair gene, mus-11, encodes a rad52 homologue and is inducible by mutagens.we isolated a neurospora crassa cdna that encodes a rad52 homologue (ncrad52) by pcr, using degenerate primers. rflp mapping demonstrated that the cloned gene is located close to the ro-4 locus on the right arm of linkage group v (lgvr). in a second experiment, we used sib selection to identify a cosmid clone containing the mus-11 gene in a n. crassa genomic library. fine-scale mapping of the mus-11 mutant showed the gene order on lgvr near ro-4 to be: ad-7 - (9.5 mu) - pab-2 (7.8 mu) - mus-11 - ...200011129042
glucose does not activate the plasma-membrane-bound h+-atpase but affects pmaa transcript abundance in aspergillus nidulans.the addition of glucose to starved cells of aspergillus nidulans increased the abundance of the pmaa transcript only transiently (15 min) and to a very low degree (1.3-fold), but strongly decreased its abundance during further incubation. this down-regulation was crea (carbon catabolite repressor protein)-dependent. glucose failed to stimulate the plasma membrane (pm)-atpase activity of a. nidulans, whereas under the same experimental conditions the activity of the enzyme from saccharomyces cere ...200011131024
evidence for the mitochondrial biosynthesis of 3r-hydroxy-5z,8z,11z,1 4z-eicosatetraenoic acid in the yeast dipodascopsis uninucleata.the biosynthesis of 3r-hydroxy-5z, 8z, 11z,14z-eicosatetraenoic acid (3r-hete) from arachidonic acid (20:4n-6) by the hyphal-forming yeast, dipodascopsis uninucleata, in cell-free enzyme extracts required coash, atp, nad+ and mg2+; 3r-hete was present as the coa derivative in enzyme extracts and its biosynthesis was associated with mitochondria. its synthesis was high from arachidonoyl-coa (15% conversion of the substrate; 22 nmol mg protein(-1) x h), but significantly higher from trans-2-arachi ...200011132180
the gdhb gene of pseudomonas aeruginosa encodes an arginine-inducible nad(+)-dependent glutamate dehydrogenase which is subject to allosteric regulation.the nad(+)-dependent glutamate dehydrogenase (nad-gdh) from pseudomonas aeruginosa pao1 was purified, and its amino-terminal amino acid sequence was determined. this sequence information was used in identifying and cloning the encoding gdhb gene and its flanking regions. the molecular mass predicted from the derived sequence for the encoded nad-gdh was 182.6 kda, in close agreement with that determined from sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme (180 kda ...200111133942
expression of the mitochondrial adp/atp carrier in escherichia coli. renaturation, reconstitution, and the effect of mutations on 10 positive residues.previously, the role of residues in the adp/atp carrier (aac) from saccharomyces cerevisiae has been studied by mutagenesis, but the dependence of mitochondrial biogenesis on functional aac impedes segregation of the mutational effects on transport and biogenesis. unlike other mitochondrial carriers, expression of the aac from yeast or mammalians in escherichia coli encountered difficulties because of disparate codon usage. here we introduce the aac from neurospora crassa in e. coli, where it is ...200111136735
the defensin peptide of the malaria vector mosquito anopheles gambiae: antimicrobial activities and expression in adult mosquitoes.a recombinant anopheles gambiae defensin peptide was used to define the antimicrobial activity spectrum against bacteria, filamentous fungi and yeast. results showed that most of the gram-positive bacterial species tested were sensitive to the recombinant peptide in a range of concentrations from 0.1 to 0.75 microm. no activity was detected against gram-negative bacteria, with the exception of some e. coli strains. growth inhibitory activity was detected against some species of filamentous fungi ...200111167093
functional analysis of the neurospora crassa pzl-1 protein phosphatase by expression in budding and fission yeast.the gene pzl-1 from the filamentous fungus neurospora crassa encodes a putative ser/thr protein phosphatase that is reminiscent of the ppz1/ppz2 and pzh1 phosphatases from saccharomyces cerevisiae and schizosaccharomyces pombe, respectively. the entire pzl-1 protein, as well as its carboxyl-terminal domain, have been expressed in escherichia coli as active protein phosphatases. to characterize its cellular role, pzl-1 was also expressed in sz. pombe and in s. cerevisiae. expression of pzl-1 in s ...200111169754
cloning and expression in phospholipid containing cultures of the gene encoding the specific phosphatidylglycerol/phosphatidylinositol transfer protein from aspergillus oryzae: evidence that the pg/pi-tp is tandemly arranged with the putative 3-ketoacyl-coa thiolase gene.the phosphatidylglycerol/phosphatidylinositol transfer protein (pg/pi-tp) is a new and original phospholipid transfer protein (pltp) isolated from the deuteromycete, aspergillus oryzae. we have isolated a genomic clone of the a. oryzae pg/pi-tp using a probe derived from the corresponding cdna and sequenced the complete gene. the dna sequence analysis revealed that pg/pi-tp gene is composed of three exons encoding a 18,823 da protein of 175 amino acids as previously described and of two introns ...200111179668
exploring infection of wheat and carbohydrate metabolism in mycosphaerella graminicola transformants with differentially regulated green fluorescent protein expression.a mycosphaerella graminicola strain transformed with the green fluorescent protein (gfp) downstream of either a carbon source-repressed promoter or a constitutive promoter was used to investigate in situ carbohydrate uptake during penetration of the fungus in wheat leaves. the promoter region of the acu-3 gene from neurospora crassa encoding isocitrate lyase was used as a carbon source-repressed promoter. the promoter region of the aspergillus nidulans gpda gene encoding glyceraldehyde-3-phospha ...200111204778
molecular control of copper homeostasis in filamentous fungi: increased expression of a metallothionein gene during aging of podospora anserina.the lifespan of the ascomycete podospora anserina was previously demonstrated to be significantly increased in a copper-uptake mutant, suggesting that copper is a potential stressor involved in degenerative processes. in order to determine whether changes in copper stress occur in the cells during normal aging of cultures, we cloned and characterized a gene coding for a component of the molecular machinery involved in the control of copper homeostasis. this gene, pamt1, is a single-copy gene tha ...200111212915
caspase-3 activates endo-exonuclease: further evidence for a role of the nuclease in apoptosis.single-strand dnase and poly raase, activities characteristic of endo-exonuclease, were co-activated in nuclear fractions of hl-60 cells by caspase-3. activation was accompanied by cleavages of large soluble polypeptides (130-185 kda) and a 65 kda inactive chromatin-associated polypeptide related to the endo-exonuclease of neurospora crassa as detected on immunoblots. the major products seen in vitro were a 77 kda soluble polypeptide and an active chromatin-associated 34 kda polypeptide. when hl ...200011225846
chemical characterization of fungal siderophores.siderophores of twenty fungi belonging to zygomycotina (5 mucorales), ascomycotina (7 aspergilli, 6 penicillia, neurospora crassa) and deuteromycotina (fusarium dimerum) were examined for their chemical nature. siderophores produced by fungi other than mucorales were all hydroxamates. mucorales produced carboxylate siderophores. catecholate type of siderophores were not detectable. hydroxamate siderophores were mostly (9 out of 15) trihydroxamates, while six were dihydroxamates. monohydroxamate ...200011233086
[differential accumulation of the new high-affinity phosphate transporter candidated gene fragment in rice roots in response to phosphorus deficiency stress].phosphate is a major constraint to crop production, and phosphate uptake in plant is mainly by high-affinity phosphate transporter under phosphate deficiency condition. using rt-pcr, a 1,178 bp phosphate transporter gene fragment ojpt1 was cloned from roots of jingxi17 (oryza sativa l. ssp. japanica) supplied with no phosphate. the comparison of this sequence with ones in genbank indicated that it shared about 70% similarity at amino acid level with other phosphate transporters in higher plants, ...200111233258
accumulation of stress and inducer-dependent plant-cell-wall-degrading enzymes during asexual development in aspergillus nidulans.determination and interpretation of fungal gene expression profiles based on digital reconstruction of expressed sequenced tags (ests) are reported. a total of 51,524 dna sequence files processed with pipeonline resulted in 9775 single and 5660 contig unique ests, 31.2% of a typical fungal transcriptome. half of the unique ests shared homology with genes in public databases, 35.8% of which are functionally defined and 64.2% are unclear or unknown. in aspergillus nidulans 86% of transcripts assoc ...200111238386
the neurospora crassa genome: cosmid libraries sorted by chromosome.a neurospora crassa cosmid library of 12,000 clones (at least nine genome equivalents) has been created using an improved cosmid vector plorist6xh, which contains a bacteriophage lambda origin of replication for low-copy-number replication in bacteria and the hygromycin phosphotransferase marker for direct selection in fungi. the electrophoretic karyotype of the seven chromosomes comprising the 42.9-mb n. crassa genome was resolved using two translocation strains. using gel-purified chromosomal ...200111238388
analysis of the pdx-1 (snz-1/sno-1) region of the neurospora crassa genome: correlation of pyridoxine-requiring phenotypes with mutations in two structural genes.we report the analysis of a 36-kbp region of the neurospora crassa genome, which contains homologs of two closely linked stationary phase genes, snz1 and sno1, from saccharomyces cerevisiae. homologs of snz1 encode extremely highly conserved proteins that have been implicated in pyridoxine (vitamin b6) metabolism in the filamentous fungi cercospora nicotianae and in aspergillus nidulans. in n. crassa, snz and sno homologs map to the region occupied by pdx-1 (pyridoxine requiring), a gene that ha ...200111238395
function of the neurospora crassa mitochondrial tyrosyl-trna synthetase in rna splicing. role of the idiosyncratic n-terminal extension and different modes of interaction with different group i introns.the neurospora crassa mitochondrial tyrosyl-trna synthetase (cyt-18 protein) promotes the splicing of group i introns by helping the intron rna fold into the catalytically active structure. the regions required for splicing include an idiosyncratic n-terminal extension, the nucleotide-binding fold domain, and the c-terminal rna-binding domain. here, we show that the idiosyncratic n-terminal region is in fact comprised of two functionally distinct parts: an upstream region consisting predominantl ...200111243805
the neurospora crassa colonial temperature-sensitive 3 (cot-3) gene encodes protein elongation factor 2.at elevated temperatures, the neurospora crassa mutant colonial, temperature-sensitive 3 (cot-3) forms compact, highly branched colonies. growth of the cot-3 strain under these conditions also results in the loss of the lower molecular weight (lmw) isoform of the ser/thr protein kinase encoded by the unlinked cot-1 gene, whose function is also involved in hyphal elongation. the unique cot-3 gene has been cloned by complementation and shown to encode translation elongation factor 2 (ef-2). as exp ...200111254137
discovery of a novel antitumor benzolactone enamide class that selectively inhibits mammalian vacuolar-type (h+)-atpases.a series of naturally occurring compounds reported recently by multiple laboratories defines a new small-molecule class sharing a unique benzolactone enamide core structure and diverse biological actions, including inhibition of growth of tumor cells and oncogene-transformed cell lines. here we show that representative members of this class, including salicylihalamide a, lobatamides a-f, and oximidines i and ii inhibit mammalian vacuolar-type (h+)-atpases (v-atpases) with unprecedented selectivi ...200111259534
the cbk1p pathway is important for polarized cell growth and cell separation in saccharomyces cerevisiae.during the early stages of budding, cell wall remodeling and polarized secretion are concentrated at the bud tip (apical growth). the cbk1 gene, encoding a putative serine/threonine protein kinase, was identified in a screen designed to isolate mutations that affect apical growth. analysis of cbk1delta cells reveals that cbk1p is required for efficient apical growth, proper mating projection morphology, bipolar bud site selection in diploid cells, and cell separation. epitope-tagged cbk1p locali ...200111259593
rcoa has pleiotropic effects on aspergillus nidulans cellular development.aspergillus nidulans rcoa encodes a member of the wd repeat family of proteins. the rcoa protein shares sequence similarity with other members of this protein family, including the saccharomyces cerevisiae tup1p and neurospora crassa rco1. tup1p is involved in negative regulation of an array of functions including carbon catabolite repression. rco1 functions in regulating pleiotropic developmental processes, but not carbon catabolite repression. in a. nidulans, deletion of rcoa (deltarcoa), a re ...200111260466
biogenesis of porin of the outer mitochondrial membrane involves an import pathway via receptors and the general import pore of the tom complex.porin, also termed the voltage-dependent anion channel, is the most abundant protein of the mitochondrial outer membrane. the process of import and assembly of the protein is known to be dependent on the surface receptor tom20, but the requirement for other mitochondrial proteins remains controversial. we have used mitochondria from neurospora crassa and saccharomyces cerevisiae to analyze the import pathway of porin. import of porin into isolated mitochondria in which the outer membrane has bee ...200111266446
expression of the avirulence gene avr9 of the fungal tomato pathogen cladosporium fulvum is regulated by the global nitrogen response factor nrf1.here we describe the role of the cladosporium fulvum nitrogen response factor 1 (nrf1) gene in regulation of the expression of avirulence gene avr9 and virulence on tomato. the nrf1 gene, which was isolated by a polymerase chain reaction-based strategy, is predicted to encode a protein of 918 amino acid residues. the protein contains a putative zinc finger dna-binding domain that shares 98% amino acid identity with the zinc finger of the major nitrogen regulatory proteins area and nit2 of asperg ...200111277429
assembly of tom6 and tom7 into the tom core complex of neurospora crassa.translocation of preproteins across the mitochondrial outer membrane is mediated by the translocase of the outer mitochondrial membrane (tom) complex. we report the molecular identification of tom6 and tom7, two small subunits of the tom core complex in the fungus neurospora crassa. cross-linking experiments showed that both proteins were found to be in direct contact with the major component of the pore, tom40. in addition, tom6 was observed to interact with tom22 in a manner that depends on th ...200111278536
actin modulates the gating of neurospora crassa vdac.vdac forms the major pathway for metabolites across the mitochondrial outer membrane. the regulation of the gating of vdac channels is an effective way to control the flow of metabolites into and out of mitochondria. here we present evidence that actin can modulate the gating process of neurospora crassa vdac reconstituted into membranes made with phosphatidylcholine. an actin concentration as low as 50 nm caused the vdac-mediated membrane conductance to drop by as much as 85% at elevated membra ...200111284205
the cryptococcus neoformans ste11alpha gene is similar to other fungal mitogen-activated protein kinase kinase kinase (mapkkk) genes but is mating type specific.partial sequence analysis of the cryptococcus neoformans matalpha mating type locus revealed the presence of a gene with substantial sequence similarity to other fungal mitogen-activated protein (map) kinase kinase kinase (mapkkk) genes. the c. neoformans gene, designated ste11alpha, showed the highest degree of similarity to the neurospora crassa nrc-1, schizosaccharomyces pombe byr2 and saccharomyces cerevisiae ste11 genes. a polymerase chain reaction-mediated sib-selection technique was succe ...200111298287
study of the role of antimicrobial glucosinolate-derived isothiocyanates in resistance of arabidopsis to microbial pathogens.crude aqueous extracts from arabidopsis leaves were subjected to chromatographic separations, after which the different fractions were monitored for antimicrobial activity using the fungus neurospora crassa as a test organism. two major fractions were obtained that appeared to have the same abundance in leaves from untreated plants versus leaves from plants challenge inoculated with the fungus alternaria brassicicola. one of both major antimicrobial fractions was purified to homogeneity and iden ...200111299350
the aspergillus nidulans gata transcription factor gene areb encodes at least three proteins and features three classes of mutation.in aspergillus nidulans, the principal transcription factor regulating nitrogen metabolism, area, belongs to the gata family of dna-binding proteins. in seeking additional gata factors, we have cloned areb, which was originally identified via a genetic screen for suppressors of area loss-of-function mutations. based on our analysis, areb is predicted to encode at least three distinct protein products. these arise from the use of two promoters, differential splicing and translation initiating at ...200111309119
characterization of an opsin gene from the ascomycete leptosphaeria maculans.an opsin gene (ops) has been characterized from leptosphaeria maculans, the ascomycete that causes black-leg disease of brassica species. this is the second opsin identified outside the archaeal and animal kingdoms. the gene encodes a predicted protein with high similarity (70.3%) and identity (53.3%) to the nop-1 opsin of another ascomycete neurospora crassa. the l. maculans opsin also has identical amino acid residues in 20 of the 22 residues in the retinal-binding pocket of archaeal opsins. o ...200111341726
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