Publications

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 ...	2001	11212915
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 ...	2001	11243805
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 ...	2001	11254137
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 ...	2000	10852494
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 ...	2000	10818103
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 ...	2000	10819445
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 ...	2000	10849201
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 ...	2000	10954093
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 ...	2000	10852489
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 ...	2000	10852493
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 ...	2000	10982393
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 ...	2000	10863006
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 ...	2000	10872226
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 ...	2000	10617646
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 ...	2000	10672443
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 ...	2000	10885790
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 ...	2000	10722661
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 ...	2000	10727211
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 ...	2000	10655216
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 ...	2000	10926509
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 ...	2000	10754257
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 ...	2000	10778749
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 ...	2000	10744987
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 ...	2000	10743568
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 ...	2000	10974123
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 ...	2000	10779395
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 ...	2000	10772469
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 ...	2000	10998244
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 ...	2000	11004444
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 ...	2000	10884222
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 ...	2000	10905354
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 ...	2000	11017770
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 ...	2000	10913868
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 ...	2000	11034781
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 ...	2000	10779483
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 ...	2000	11057446
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 ...	2000	11073919
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 ...	2000	10919380
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 ...	2000	10794177
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 ...	2000	10817227
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 ...	2000	10931877
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 ...	2000	10600685
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 ...	2000	10641037
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 ...	2000	10731431
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 ...	2000	10736021
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 ...	2000	10739348
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 ...	2000	10675602
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 ...	2000	10712689
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 ...	2000	10783241
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 ...	2000	10784039
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 ...	2000	10656585
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 ...	2000	10600684
mips: a database for genomes and protein sequences.	the munich information center for protein sequences (mips-gsf), martinsried, near munich, germany, continues its longstanding tradition to develop and maintain high quality curated genome databases. in addition, efforts have been intensified to cover the wealth of complete genome sequences in a systematic, comprehensive form. bioinformatics, supporting national as well as european sequencing and functional analysis projects, has resulted in several up-to-date genome-oriented databases. this repo ...	2000	10592176
mitop, the mitochondrial proteome database: 2000 update.	mitop (http://www.mips.biochem.mpg.de/proj/medgen/mitop/) is a comprehensive database for genetic and functional information on both nuclear- and mitochondrial-encoded proteins and their genes. the five species files--saccharomyces cerevisiae, mus musculus, caenorhabditis elegans, neurospora crassa and homo sapiens--include annotated data derived from a variety of online resources and the literature. a wide spectrum of search facilities is given in the overlapping sections 'gene catalogues', 'pr ...	2000	10592209
multiple intracellular peptidases in neurospora crassa.	neurospora crassa possesses multiple intracellular peptidases which display overlapping substrate specificities. they were readily detected by an in situ staining procedure for peptidases separated in polyacrylamide gels, within which the auxilliary enzyme, l-amino acid oxidase, was immobilized. eleven different intracellular peptidases were identified by electrophoretic separation and verified by their individual patterns of substrate specificities. most peptide substrates tested were hydrolyze ...	2000	86534
purification and characterization of 3-dehydroshikimate dehydratase, an enzyme in the inducible quinic acid catabolic pathway of neurospora crassa.	3-dehydroshikimate dehydratase catalyzes the third reaction in the inducible quinic acid catabolic pathway of neurospora crassa and is encoded in the qa-4 gene of the qa gene cluster. as part of continuing genetic and biochemical studies concerning the organization and regulation of this gene cluster, 3-dehydroshikimate dehydratase has been purified and characterized biochemically. the enzyme was purified 1650-fold using the following techniques: 1) (nh4)2so4 fractionation; 2) ion exchange chrom ...	2000	149131
does cycloheximide-induced loss of phosphate uptake activity in neurospora crassa reflect rapid turnover?	addition of cycloheximide to neurospora crassa germlings growing in liquid medium caused an exponential loss of phosphate uptake activity (half-life, ca. 2 h). no loss of activity resulted when germlings were resuspended, at the time of cycloheximide addition, in medium of a substantially lower phosphate concentration. it is concluded that the phosphate uptake systems are not subject to rapid turnover.	2000	149111
an unstable allele of the am locus of neurospora crassa.	the mutant strain am126 was isolated, using the direct selection procedure, after nitrous acid mutagenesis. it produced neither measurable nadp-dependent glutamate dehydrogenase (gdh) nor immunologically cross-reacting material. that the am126 strain produced some form of gdh product was shown by the fact that it complemented several other am mutant strains. the gdh formed by complementation between am126 and each of two other am mutants was relatively thermolabile, but could not be distinguishe ...	2000	94569
isolation and characterization of a mitochondrial d-amino acid oxidase from neurospora crassa.	d-amino acid oxidase (ec 1.4.3.3) activity in homogenates of neurospora crassa strain sy7a was found to sediment with the mitochondrial fraction. digitonin fractionation studies on purified mitochondria have indicated a matrix localization of the enzyme. additionally, a peroxidase (ec 1.11.1.7) activity, which may remove hydrogen peroxide formed as a product of d-amino acid oxidation, was also found in the mitochondrial matrix. partial purification (20- to 30-fold) of the mitochondrial d-amino a ...	2000	13919
cellulase of neurospora crassa.	mycelia and ungerminated conidia of neurospora crassa were found to secrete extracellular endocellulase (ec 3.2.1.4). a simple induction system of potassium phosphate buffer (ph 6.0) plus inducer relied on the internal metabolic reserves of conicia or mycelia to provide energy and substrates for protein synthesis. buffer concentration for optimum enzyme production was 100 mm, but at higher buffer concentrations enzyme production was inhibited. cellobiose was clearly the best inducer, with an opt ...	2000	15975
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 ...	2000	11003665
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 ...	2000	11024266
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 ...	2000	11131024
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 ...	2000	11132180
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 ...	2000	11057443
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 - ...	2000	11129042
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 ...	2000	10955907
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 ...	2000	10953880
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 ...	2000	10880472
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 ...	2000	10882535
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 ...	2000	10862611
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 ...	2000	11013078
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 ...	2000	11016845
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 ...	2000	11225846
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 ...	2000	11233086
functional characterization of the conserved "glk" motif in mitochondrial porin from neurospora crassa.	mitochondrial porin facilitates the diffusion of small hydrophilic molecules across the mitochondrial outer membrane. despite low sequence similarity among porins from different species, a "glycine-leucine-lysine" (glk) motif is conserved in mitochondrial and neisseria porins. to investigate the possible roles of these conserved residues, including their hypothesized participation in atp binding by the protein, we replaced the lysine residue of the glk motif of neurospora crassa porin with gluta ...	2000	15254370
hex-1, a gene unique to filamentous fungi, encodes the major protein of the woronin body and functions as a plug for septal pores.	we have identified a gene, named hex-1, that encodes the major protein in the hexagonal crystals, or woronin bodies, of neurospora crassa. analysis of a strain with a null mutation in the hex-1 gene showed that the septal pores in this organism were not plugged when hyphae were damaged, leading to extensive loss of cytoplasm. when grown on agar plates containing sorbose, the hex-1(-) strain showed extensive lysis of hyphal tips. the hex-1 protein was predicted to be 19,125 da. analysis of the n- ...	2000	11273682
circadian rhythms in neurospora crassa: lipid deficiencies restore robust rhythmicity to null frequency and white-collar mutants.	the conidiation rhythm in the fungus neurospora crassa is a model system for investigating the genetics of circadian clocks. null mutants at the frq (frequency) locus (frq(9) and frq(10)) make no functional frq gene products and are arrhythmic under standard conditions. the white-collar strains (wc-1 and wc-2) are insensitive to most effects of light, and are also arrhythmic. all three genes are proposed to be central components of the circadian oscillator. we have been investigating two mutants ...	2000	10618405
disruption of vma-1, the gene encoding the catalytic subunit of the vacuolar h(+)-atpase, causes severe morphological changes in neurospora crassa.	by using the process of repeat-induced point mutation (selker, e. u., and garrett, p. w. (1988) proc. natl. acad. sci. u. s. a. 85, 6870-6874), we inactivated vma-1, the gene encoding subunit a of the v-atpase of neurospora crassa. two vma-1 mutant strains were characterized. one was mutated at multiple sites, did not make a protein product, and produced spores that only rarely germinated. the other had four point mutations, made a protein product, and produced viable spores. neither strain had ...	2000	10617601
a neurospora crassa arp1 mutation affecting cytoplasmic dynein and dynactin localization.	of the actin-related proteins, arp1 is the most similar to conventional actin, and functions solely as a component of the multisubunit complex dynactin. dynactin has been identified as an activator of the microtubule-associated motor cytoplasmic dynein. the role of arp1 within dynactin is two-fold: (1) it serves as a structural scaffold protein for other dynactin subunits; and (2) it has been proposed to link dynactin, and thereby dynein, with membranous cargo via interaction with spectrin. usin ...	2000	11129047
ph homeostasis of the circadian sporulation rhythm in clock mutants of neurospora crassa.	the influence of environmental (extracellular) ph on the sporulation rhythm in neurospora crassa was investigated for wild-type (frq+) and the mutants chr, frq1, frq7, and frq8. in all mutants, including wild type, the growth rate was found to be influenced strongly by extracellular ph in the range 4-9. on the other hand, for the same ph range, the period length of the sporulation rhythm is little influenced in wild type, chr, and frq1. a loss of ph homeostasis of the period, however, was observ ...	2000	11128290
heat shock protein 80 of neurospora crassa: sequence analysis of the gene and expression during the asexual phase.	heat shock protein 80 (hsp80) of neurospora crassa, a member of the stress-90 protein family, is a cytosolic molecular chaperone that interacts directly with hsp70 to form a hetero-oligomeric complex. the complete nucleotide sequence of the gene encoding this protein, along with the 5'- and 3'-flanking dna, is reported. the coding sequence is interrupted by two introns, 61 and 30 nucleotides, respectively, in length. the deduced amino acid sequence corresponds to a 695-residue polypeptide with a ...	2000	11109485
in vivo chitin-cadmium complexation in cell wall of neurospora crassa.	fungal cell wall, mainly composed of chitin, an n-acetylglucosamine polymer, is known to participate in heavy metal detoxification. in the present study, an effort was made to elucidate the sites involved in complexation of cadmium by the chitin material of cell wall of neurospora crassa. based on the results of physical techniques, such as solid-state 13c-nmr, x-ray diffraction, ir and molecular modeling, a structure was proposed for the chitin-cadmium complex. the ring and c-3 hydroxyl oxygens ...	2000	11099854
crystallization, structure and dynamics of the proton-translocating p-type atpase.	large single three-dimensional crystals of the dodecylmaltoside complex of the neurospora crassa plasma membrane h(+)-atpase (h(+) p-atpase) can be grown in polyethylene-glycol-containing solutions optimized for moderate supersaturation of both the protein surfaces and detergent micellar region. large two-dimensional h(+) p-atpase crystals also grow on the surface of such mixtures and on carbon films located at such surfaces. electron crystallographic analysis of the two-dimensional crystals gro ...	2000	10600683
cellular role of the v-atpase in neurospora crassa: analysis of mutants resistant to concanamycin or lacking the catalytic subunit a.	vacuolar atpases (v-atpases) are large complex enzymes that are structural and mechanistic relatives of f(1)f(o)-atpases. they hydrolyze atp and pump protons across membranes to hyperpolarize membranes and, often, to acidify cellular compartments. the proton gradients generated are used to drive the movement of various compounds across membranes. v-atpases are found in membranes of archaebacteria and some eubacteria, in various components of the endomembrane system of all eukaryotes and in the p ...	2000	10600678
a conserved mechanism for post-transcriptional gene silencing?	proteins with homology to rna-directed rna polymerases function in post-transcriptional gene silencing: in quelling in the fungus neurospora crassa, rnai in the nematode caenorhabditis elegans, and co-suppression in the mustard plant arabidopsis thaliana. these findings are consistent with a conserved mechanism operating in these diverse species.	2000	11178243
repeat-induced point mutation (rip) in crosses with wild-isolated strains of neurospora crassa: evidence for dominant reduction of rip.	seventy-one wild-isolated strains of neurospora crassa were examined for their ability to support repeat-induced point mutation (rip) in the erg-3 locus. rip was exceptionally inefficient but detectable in crosses with the strain fgsc 430 from adiopodoume, ivory coast. we could find no consistent differences in ascospore yields when wild isolates identified as "low-rip" or "high-rip" strains were crossed with strains bearing the segmental duplication dp(iiir > [i; ii])ar17. this suggested that r ...	2000	11170738
an osmotic-remedial, temperature-sensitive mutation in the allosteric activity site of ribonucleotide reductase in neurospora crassa.	an osmotic-remedial, temperature-sensitive conditional mutant (un-24) was generated by repeat induced point mutation (rip) from a cross between a wild-type n. crassa strain and a strain carrying a approximately 250-kb duplication of the left arm of linkage group ii (lgii). the mutation was mapped to the duplicated segment, within 2.6 map units of the heterokaryon incompatibility locus het-6. dna transformation identified a 3.75-kb fragment that complemented the temperature-sensitive phenotype. a ...	2000	10660063
characterization of an unusual trna-like sequence found inserted in a neurospora retroplasmid.	we characterized an unusual trna-like sequence that had been found inserted in suppressive variants of the mitochondrial retroplasmid of neurospora intermedia strain varkud. we previously identified two forms of the trna-like sequence, one of 64 nt (trl-64) and the other of 78 nt (trl-78) containing a 14-nt internal insertion in the anticodon stem at a position expected for a nuclear trna intron. here, we show that trl-78 is encoded in varkud mitochondrial (mt)dna within a 7 kb sequence that is ...	2000	10710417
ego-1 is related to rna-directed rna polymerase and functions in germ-line development and rna interference in c. elegans.	cell-fate determination requires that cells choose between alternative developmental pathways. for example, germ cells in the nematode worm caenorhabditis elegans choose between mitotic and meiotic division, and between oogenesis and spermatogenesis. germ-line mitosis depends on a somatic signal that is mediated by a notch-type signaling pathway. the ego-1 gene was originally identified on the basis of genetic interactions with the receptor in this pathway and was also shown to be required for o ...	2000	10704412
nadh dehydrogenase in neurospora crassa contains myristic acid covalently linked to the nd5 subunit peptide.	the mitochondrial, proton-pumping nadh:ubiquinone oxidoreductase consists of at least 35 subunits whose synthesis is divided between the cytosol and mitochondria; this complex i catalyzes the first steps of mitochondrial electron transfer and proton translocation. radiolabel from [(3)h]myristic acid was incorporated by neurospora crassa into the mitochondrial-encoded, approximately 70 kda nd5 subunit of nadh dehydrogenase, as shown by immunoprecipitation. this myristate apparently was linked to ...	2000	10699461
cryo-electron tomography of neurospora mitochondria.	cryo-electron tomography was used to study the structural organization of whole frozen-hydrated mitochondria from neurospora crassa. unlike mitochondria from many other species and tissues, in this case the cristae form a three-dimensional network of interconnected lamellae. basically, the three-dimensional structure of ice-embedded mitochondria from this species is consistent with previous descriptions of mitochondria prepared by chemical fixation and resin embedding. nonetheless, ice-embedded ...	2000	10675296
use of an isocitrate lyase promoter-gfp fusion to monitor carbon metabolism of the plant pathogen tapesia yallundae during infection of wheat.	abstract green fluorescent protein (gfp) has been used as a vital marker in a variety of species. here, we present the use of a gfp-promoter fusion to visualize carbon metabolism in a pathogenic fungus during growth on defined medium and during infection of plants. isocitrate lyase (icl), a key enzyme in carbon metabolism, is tightly regulated at the transcriptional level, with high levels of expression during 2-carbon growth and no expression during growth on glucose. a gfp-icl promoter fusion ...	2000	20572972
short tpa-rich segments of the zeta-eta region induce dna methylation in neurospora crassa.	the mechanisms that establish dna methylation in eukaryotes are poorly understood. in principle, methylation in a particular chromosomal region may reflect the presence of a "signal" that recruits methylation, the absence of a signal that prevents methylation, or both. experiments were carried out to address these possibilities for the 1.6 kb zeta-eta (zeta-eta) region, a relict of repeat-induced point mutation (rip) in the fungus neurospora crassa. the zeta-eta region directs its own de novo me ...	2000	10873464
the cyanide-resistant alternative oxidases from the fungi pichia stipitis and neurospora crassa are monomeric and lack regulatory features of the plant enzyme.	both plant and fungal mitochondria have cyanide-resistant alternative oxidases that use reductant from the mitochondrial ubiquinone pool to reduce oxygen to water in a reaction that conserves no energy for atp synthesis. the dimeric plant alternative oxidase is relatively inactive when its subunits are linked by a disulfide bond. when this bond is reduced, the enzyme can then be stimulated by its activators, alpha-keto acids. a cys in the n-terminal section of the protein is responsible for both ...	2000	10860541
sn-1,2-diacylglycerol levels in the fungus neurospora crassa display circadian rhythmicity.	the fungus neurospora crassa is a model organism for investigating the biochemical mechanism of circadian (daily) rhythmicity. when a choline-requiring strain (chol-1) is depleted of choline, the period of the conidiation rhythm lengthens. we have found that the levels of sn-1,2-diacylglycerol (dag) increase in proportion to the increase in period. other clock mutations that change the period do not affect the levels of dag. membrane-permeant dags and inhibitors of dag kinase were found to furth ...	2000	10859307
respiratory chain complex i is essential for sexual development in neurospora and binding of iron sulfur clusters are required for enzyme assembly.	we have cloned and disrupted in vivo, by repeat-induced point mutations, the nuclear gene coding for an iron sulfur subunit of complex i from neurospora crassa, homologue of the mammalian tyky protein. analysis of the obtained mutant nuo21.3c revealed that complex i fails to assemble. the peripheral arm of the enzyme is disrupted while its membrane arm accumulates. furthermore, mutated 21.3c-kd proteins, in which selected cysteine residues were substituted with alanines or serines, were expresse ...	2000	11014810
characterization of an unusual trna-like sequence found inserted in a neurospora retroplasmid.	we characterized an unusual trna-like sequence that had been found inserted in suppressive variants of the mitochondrial retroplasmid of neurospora intermedia strain varkud. we previously identified two forms of the trna-like sequence, one of 64 nt (trl-64)and the other of 78 nt (trl-78) containing a 14-nt internal insertion in the anticodon stem at a position expected for a nuclear trna intron. here, we show that trl-78 is encoded in varkud mitochondrial (mt)dna within a 7 kb sequence that is n ...	2000	11001704
cellular distribution of cot1 kinase in neurospora crassa.	the neurospora crassa cot-1 gene encodes a ser/thr protein kinase, which is involved in hyphal elongation. many vacuoles, abnormally shaped mitochondria, and nuclei, along with differences in the structure of the cell wall and hyphal septa, were observed in hyphae of the cot-1 mutant shortly after a shift to the restrictive temperature. immunolocalization experiments indicated that cot1 was associated with the cytoplasmic membrane; cot1 was also detected in the cytoplasm. the membrane-associated ...	2000	10955908
a putative spectrin-containing membrane skeleton in hyphal tips of neurospora crassa.	the apical plasma membrane (pm) is important in hyphal tip growth, where it may regulate tip extensibility via its association with an appropriate membrane skeleton (ms). by cell fractionation and immunocytochemistry we show that proteins with characteristics of actin, spectrin, and integrin are associated in a ms-like manner with the pm of neurospora crassa hyphae. the spectrin-like protein in particular is highly concentrated at the pm in the region of maximum apical expansion. this protein sh ...	2000	10955906