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convergent evolution of hydroxylation mechanisms in the fungal kingdom: molybdenum cofactor-independent hydroxylation of xanthine via alpha-ketoglutarate-dependent dioxygenases.the xanthine oxidases and dehydrogenases are among the most conserved enzymes in all living kingdoms. they contain the molybdopterin cofactor moco. we show here that in the fungi, in addition to xanthine dehydrogenase, a completely different enzyme is able to catalyse the oxidation of xanthine to uric acid. in aspergillus nidulans this enzyme is coded by the xana gene. we have cloned the xana gene and determined its sequence. a deletion of the gene has the same phenotype as the previously known ...200515948966
evaluation of phenylaminopyrimidines as antifungal protein kinase inhibitors.the effects of diverse phenylaminopyrimidines (pap), namely pap-pyridines (type a), pap-pyrazoles (type b) and pap-thiazoles (type c), on neurospora crassa shear & dodge has been investigated. the results revealed that type a strongly inhibit the in vitro growth of n crassa, whereas types b and c are much less active. a significant correlation was observed between the neurospora growth inhibition and the intrinsic activity of type a compounds on the cyclin-dependent protein kinase p34(cdc2) of s ...200515952247
neurospora strains harboring mitochondrial disease-associated mutations in iron-sulfur subunits of complex i.we subjected the genes encoding the 19.3-, 21.3c-, and 51-kda iron-sulfur subunits of respiratory chain complex i from neurospora crassa to site-directed mutagenesis to mimic mutations in human complex i subunits associated with mitochondrial diseases. the v135m substitution was introduced into the 19.3-kda cdna, the p88l and r111h substitutions were separately introduced into the 21.3c-kda cdna, and the a353v and t435m alterations were separately introduced into the 51-kda cdna. the altered cdn ...200515956670
functions of the small proteins in the tom complex of neurospora crasssa.the tom (translocase of the outer mitochondrial membrane) complex of the outer mitochondrial membrane is required for the import of proteins into the organelle. the core tom complex contains five proteins, including three small components tom7, tom6, and tom5. we have created single and double mutants of all combinations of the three small tom proteins of neurospora crassa. analysis of the mutants revealed that tom6 plays a major role in tom complex stability, whereas tom7 has a lesser role. mut ...200515987740
antifungal coating by biofunctionalized polyelectrolyte multilayered films.the surface of medical devices is a common site of bacterial and fungal adhesion, first step to the constitution of a resistant biofilm leading frequently to chronic infections. in order to prevent such complications, several physical and chemical modifications of the device surface have been proposed. here, we experiment a new type of topical antifungal coating using the layer-by-layer technique. the nanometric multilayer film obtained by this technique is functionalized by the insertion of a c ...200515992921
oxygen and hydrogen peroxide enhance light-induced carotenoid synthesis in neurospora crassa.previously, we found that intracellular reactive oxygen species (ros) affect photomorphogenesis in neurospora crassa. in this study, we investigated the physiological roles of ros in the response to light and found that the exposure of mycelia to air was important for the light-induced carotenogenesis. mycelia treated with a high concentration of o(2) gas and h(2)o(2) to release ros showed an enhancement of light-induced carotenoid accumulation and the expression of gene related to light-inducib ...200516004990
the 29.9 kda subunit of mitochondrial complex i is involved in the enzyme active/de-active transitions.mitochondrial respiratory chain complex i undergoes transitions from active to de-activated forms. we have investigated the phenomenon in sub-mitochondrial particles from neurospora crassa wild-type and a null-mutant lacking the 29.9 kda nuclear-coded subunit of complex i. based on enzymatic activities, genetic crosses and analysis of mitochondrial proteins in sucrose gradients, we found that about one-fifth of complex i with catalytic properties similar to the wild-type enzyme is assembled in t ...200516005890
mass flow and pressure-driven hyphal extension in neurospora crassa.mass flow of cytoplasm in neurospora crassa trunk hyphae was directly confirmed by injecting oil droplets into the hyphae. the droplets move in a manner similar to cytoplasmic particles and vacuoles within the hyphae. the direction of mass flow is towards the growing hyphal tips at the colony edge. based on flow velocities (about 5 microm s(-1)), hyphal radius and estimates of cytoplasm viscosity, the reynolds number is about 10(-4), indicating that mass flow is laminar. therefore, the poiseulle ...200516079346
[photoreceptor apparatus of a fungus neurospora crassa].light governs living functions of ascomycete fungus neurospora crassa by controlling expression of the genes responsible for differentiation of reproductive structures, synthesis of secondary metabolites and the circadian oscillator activity. illumination also influences electrogenic processes in cell membrane and the activity and molecular organization of some enzymes. the major but, probably, not the sole photoreceptor pigment in neurospora cells is wcc, a heterodimeric complex formed by pas-d ...200616083009
methylation of histone h3 lysine 36 is required for normal development in neurospora crassa.the set domain is an evolutionarily conserved domain found predominantly in histone methyltransferases (hmts). the neurospora crassa genome includes nine set domain genes (set-1 through set-9) in addition to dim-5, which encodes a histone h3 lysine 9 hmt required for dna methylation. we demonstrate that neurospora set-2 encodes a histone h3 lysine 36 (k36) methyltransferase and that it is essential for normal growth and development. we used repeat induced point mutation to make a set-2 mutant (s ...200516087750
temperature-sensitive, osmotic-remedial mutants of neurospora crassa: osmotic pressure induced alterations of enzyme stability.three temperature-sensitive, adenine-requiring mutants of neurospora crassa were found to be osmotic-remedial when non-penetrating solutes were used to increase the osmolarity of the growth medium. the affected enzyme (adenylosuccinase) from one of the mutants (ad-4, 44206t) was found to have higher levels of activity when the organism was grown at non-permissive temperatures under osmotic-remedial conditions than when it was grown with adenine as a nutritional supplement. the enzyme synthesized ...197516094964
transcription of repetitive dna in neurospora crassa.repeated dna sequences of neurospora crassa were isolated and characterized. approximately 10-12% of n. crassa dna sequence were repeated, of which 7.3% were found to be transcribed in mid-log phase of mycelial growth as measured by dna:rna hybridization. it is suggested that part of repetitive dna transcripts in n. crassa were mitochondrial and part were nuclear dna. most of the nuclear repeated dnas, however, code for rrna and trna in n. crassa.197516094973
influence of edeine on intergenic and interallelic recombination in neurospora crassa.the effect of edeine and the mutation edr-2 to edeine resistance on genetic recombination in neurospora crassa was investigated. for this purpose crosses between pairs of edeine sensitive and edeine resistant strains respectively were set up without or in the presence of the drug (0-750 microg/ml). the genetic markers ylo-1, ad-1, pan-2 (b3 and b5) and tryp-2, all on linkage group vi, were used for scoring recombinants. these were ad+, tryp+ (intergenic recombination) and pan+ (interallelic reco ...197516094998
the e-hook of tubulin interacts with kinesin's head to increase processivity and speed.kinesins are dimeric motor proteins that move processively along microtubules. it has been proposed that the processivity of conventional kinesins is increased by electrostatic interactions between the positively charged neck of the motor and the negatively charged c-terminus of tubulin (e-hook). in this report we challenge this anchoring hypothesis by studying the motility of a fast fungal kinesin from neurospora crassa (nckin). nckin is highly processive despite lacking the positive charges in ...200516100283
molecular mechanism of temperature sensing by the circadian clock of neurospora crassa.expression levels and ratios of the long (l) and short (s) isoforms of the neurospora circadian clock protein frequency (frq) are crucial for temperature compensation of circadian rhythms. we show that the ratio of l-frq versus s-frq is regulated by thermosensitive splicing of intron 6 of frq, a process removing the translation initiation site of l-frq. thermosensitivity is due to inefficient recognition of nonconsensus splice sites at elevated temperature. the temperature-dependent accumulation ...200516107616
comparative genomics and disorder prediction identify biologically relevant sh3 protein interactions.protein interaction networks are an important part of the post-genomic effort to integrate a part-list view of the cell into system-level understanding. using a set of 11 yeast genomes we show that combining comparative genomics and secondary structure information greatly increases consensus-based prediction of sh3 targets. benchmarking of our method against positive and negative standards gave 83% accuracy with 26% coverage. the concept of an optimal divergence time for effective comparative ge ...200516110343
paracoccidioides brasiliensis translation and protein fate machineries revealed by functional genome analysis.the translational and post-translational modification machineries of paracoccidioides brasiliensis were assessed by means of comparative analyses of pbaests (p. brasiliensis assembled expressed sequence tags) with sequences deposited on different databases. of the 79 sequences corresponding to cytosolic ribosomal proteins, we were able to find 78 in the p. brasiliensis transcriptome. nineteen of the 27 saccharomyces cerevisiae genes related to translation initiation were also found. all eukaryot ...200516110446
the ergosterol biosynthesis pathway, transporter genes, and azole resistance in aspergillus fumigatus.the continuous use of triazoles can result in the development of drug resistance. azole-resistant clinical isolates, spontaneous and induced mutants of aspergillus fumigatus have been documented. the azoles block the ergosterol biosynthesis pathway by inhibiting the enzyme 14-alpha-demethylase, product of the cyp51. fungal azole resistance involves both amino acid changes in the target site that alter drug-target interactions and those that decrease net azole accumulation. the reduced intracellu ...200516110826
transcripts and transcript-binding proteins in mitochondria of neurospora crassa.we analyzed expression elements of three disparate groups of mitochondrial genes in neurospora crassa, apocytochrome b (cob), cytochrome c oxidase 1 (cox1), and the clustered atp8-atp6-mtatp9-cox2. to identify promoter sequences we employed the published n. crassa consensus sequence for cob and rrna genes, and we found closely related sequences within the 5'-regions of both cox1 and the atp8-cox2 transcriptional units. we determined that the mature cox1 rna includes two flanking unassigned readi ...200316120332
deciphering the model pathogenic fungus cryptococcus neoformans.cryptococcus neoformans is a basidiomycete fungal pathogen of humans that has diverged considerably from other model fungi such as neurospora crassa, aspergillus nidulans, saccharomyces cerevisiae and the common human fungal pathogen candida albicans. the recent completion of the genome sequences of two related c. neoformans strains and the ongoing genome sequencing of three other divergent cryptococcus strains with different virulence phenotypes and environmental distributions should improve ou ...200516132036
biotransformation of racemic diisophorone by cephalosporium aphidicola and neurospora crassa.racemic diisophorone (500 mg) was converted by cephalosporium aphidicola and neurospora crassa over 10 days at 25 degrees c to 8beta-hydroxydiisophorone in yields of 10% (52 mg) and 20% (103 mg), respectively. the structure was established by ir, specific rotation, mass spectral, 1d and 2d-nmr studies.200516132845
rna silencing in fungi: mechanisms and applications.two rna silencing-related phenomena, quelling and meiotic silencing by unpaired dna (msud) have been identified in the fungus neurospora crassa. similar to the case with the sirna and mirna pathways in drosophila, different sets of protein components including rna-dependent rna polymerase, argonaute and dicer, are used in the quelling and msud pathways. orthologs of the rna silencing components are found in most, but not all, fungal genomes currently available in the public databases, indicating ...200516137680
circadian rhythms in neurospora crassa: clock gene homologues in fungi.computer-based analysis of a total of 17 filamentous fungal and yeasts genomes has shown: (1) homologues of frq, wc-1, wc-2, and vvd, key gene components of the neurospora crassa clock, are present in magnaporthe grisea, gibberella zeae, and podospora anserina, suggesting an frq-based oscillator in these organisms; (2) some fungal species that are more distantly related to neurospora, such as rhizopus oryzae do not appear to have frq homologues; (3) many fungal species that do not appear to cont ...200516154782
temperature-modulated alternative splicing and promoter use in the circadian clock gene frequency.the expression of frequency, a central component of the circadian clock in neurospora crassa, shows daily cycles that are exquisitely sensitive to the environment. two forms of frq that differ in length by 99 amino acids, lfrq and sfrq, are synthesized from alternative initiation codons and the change in their ratio as a function of temperature contributes to robust rhythmicity across a range of temperatures. we have found frq expression to be surprisingly complex, despite our earlier prediction ...200516195340
[riboswitches].riboswitches are rna structures able to bind small molecules and regulate gene expression at both, transcriptional and translational level. they are present in a wide variety of bacterial species. in bacillus subtilis more than 2% of the genome is regulated by riboswitches. metabolite binding is highly specific and can be provided in cooperative manner. several riboswitches has been identified and characterized to be specific for flavin mononucleotide, thiamine, thiamine pyrophosphate, adenosylc ...200516209348
preprotein translocase of the outer mitochondrial membrane: reconstituted tom40 forms a characteristic tom pore.tom40 is the central pore-forming component of the translocase of the outer mitochondrial membrane (tom complex). different views exist about the secondary structure and electrophysiological characteristics of tom40 from saccharomyces cerevisiae and neurospora crassa. we have directly compared expressed and renatured tom40 from both species and find a high content of beta-structure in circular dichroism measurements in agreement with refined secondary structure predictions. the electrophysiologi ...200516213519
human mitochondrial complex i assembly is mediated by ndufaf1.complex i (nadh:ubiquinone oxidoreductase) is the largest multiprotein enzyme of the oxidative phosphorylation system. its assembly in human cells is poorly understood and no proteins assisting this process have yet been described. a good candidate is ndufaf1, the human homologue of neurospora crassa complex i chaperone cia30. here, we demonstrate that ndufaf1 is a mitochondrial protein that is involved in the complex i assembly process. modulating the intramitochondrial amount of ndufaf1 by kno ...200516218961
growth defect and mutator phenotypes of recq-deficient neurospora crassa mutants separately result from homologous recombination and nonhomologous end joining during repair of dna double-strand breaks.recq helicases function in the maintenance of genome stability in many organisms. the filamentous fungus neurospora crassa has two recq homologs, qde3 and recq2. we found that the qde-3 recq2 double mutant showed a severe growth defect. the growth defect was alleviated by mutation in mei-3, the homolog of yeast rad51, which is required for homologous recombination (hr), suggesting that hr is responsible for this phenotype. we also found that the qde-3 recq2 double mutant showed a mutator phenoty ...200616219790
endocytosis in the plant-pathogenic fungus ustilago maydis.filamentous fungi are an important group of tip-growing organisms, which include numerous plant pathogens such as magnaporthe grisea and ustilago maydis. despite their ecological and economical relevance, we are just beginning to unravel the importance of endocytosis in filamentous fungi. most evidence for endocytosis in filamentous fungi is based on the use of endocytic tracer dyes that are taken up into the cell and delivered to the vacuole. moreover, genomewide screening for candidate genes i ...200516231103
cloning and sequencing of the chromosomal dna and cdna encoding the mitochondrial citrate synthase of aspergillus niger wu-2223l.the complementary dna (cdna) and chromosomal dna encoding the citrate synthase (ec 4.1.3.7) gene (cit1) of aspergillus niger wu-2223l, a citric acid-producing strain, were cloned. synthetic oligonucleotide primers were designed according to the amino acid sequences of already known eukaryotic citrate synthases and the codon bias of a. niger genes. the 920-bp dna fragment was amplified by polymerase chain reaction with these primers using chromosomal dna of wu-2223l as a template, and was employe ...199916232605
synchronizing the neurospora crassa circadian clock with the rhythmic environment.the metronomic predictability of the environment has elicited strong selection pressures for the evolution of endogenous circadian clocks. circadian clocks drive molecular and behavioural rhythms that approximate the 24 h periodicity of our environment. found almost ubiquitously among phyla, circadian clocks allow preadaptation to rhythms concomitant with the natural cycles of the earth. cycles in light intensity and temperature for example act as important cues that couple circadian clocks to t ...200516246018
the pas/lov protein vivid supports a rapidly dampened daytime oscillator that facilitates entrainment of the neurospora circadian clock.a light-entrainable circadian clock controls development and physiology in neurospora crassa. existing simple models for resetting based on light pulses (so-called nonparametric entrainment) predict that constant light should quickly send the clock to an arrhythmic state; however, such a clock would be of little use to an organism in changing photoperiods in the wild, and we confirm that true, albeit dampened, rhythmicity can be observed in extended light. this rhythmicity requires the pas/lov p ...200516264193
the gene for a lectin-like protein is transcriptionally activated during sexual development, but is not essential for fruiting body formation in the filamentous fungus sordaria macrospora.the filamentous fungus sordaria macrospora forms complex three-dimensional fruiting bodies called perithecia that protect the developing ascospores and ensure their proper discharge. in previous microarray analyses, several genes have been identified that are downregulated in sterile mutants compared to the wild type. among these genes was tap1 (transcript associated with perithecial development), a gene encoding a putative lectin homolog.200516266439
[study on xylose fermentation by neurospora crassa].the influence of oxygen limitation and medium initial ph on xylose fermentation by neurospora crassa as 3.1602 was investigated. n. crassa as 3.1602 has high ability of xylose fermentation producing ethanol and xylitol. oxygen limitation has big influence on ethanol and xylitol production. the maximum conversion rate of ethanol was 63.2% obtained under semi-aerobic conditions. the maximum conversion rate of xylitol was 31.8% obtained under micro-aerobic conditions. xylitol accumulation is behind ...200316276921
group iii histidine kinase is a positive regulator of hog1-type mitogen-activated protein kinase in filamentous fungi.we previously reported that the group iii histidine kinase dic1p in the maize pathogen cochliobolus heterostrophus is involved in resistance to dicarboximide and phenylpyrrole fungicides and in osmotic adaptation. in addition, exposure to the phenylpyrrole fungicide fludioxonil led to improper activation of hog1-type mitogen-activated protein kinases (mapks) in some phytopathogenic fungi, including c. heterostrophus. here we report, for the first time, the relationship between the group iii hist ...200516278449
a rho-type gtpase, rho-4, is required for septation in neurospora crassa.proteins in the rho family are small monomeric gtpases primarily involved in polarization, control of cell division, and reorganization of cytoskeletal elements. phylogenetic analysis of predicted fungal rho proteins suggests that a new rho-type gtpase family, whose founding member is rho4 from the archiascomycete schizosaccharomyces pombe, is involved in septation. s. pombe rho4delta mutants have multiple, abnormal septa. in contrast to s. pombe rho4delta mutants, we show that strains containin ...200516278458
molecular mechanism of light responses in neurospora: from light-induced transcription to photoadaptation.blue light regulates many molecular and physiological activities in a large number of organisms. in neurospora crassa, a eukaryotic model system for studying blue-light responses, the transcription factor and blue-light photoreceptor white collar-1 (wc-1) and its partner wc-2 are central to blue-light sensing. neurospora's light responses are transient, that is, following an initial acute phase of induction, light-regulated processes are down-regulated under continuous illumination, a phenomenon ...200516287715
long-oligomer microarray profiling in neurospora crassa reveals the transcriptional program underlying biochemical and physiological events of conidial germination.to test the inferences of spotted microarray technology against a biochemically well-studied process, we performed transcriptional profiling of conidial germination in the filamentous fungus, neurospora crassa. we first constructed a 70 base oligomer microarray that assays 3366 predicted genes. to estimate the relative gene expression levels and changes in gene expression during conidial germination, we analyzed a circuit design of competitive hybridizations throughout a time course using a baye ...200516287898
proteome analysis of mitochondrial outer membrane from neurospora crassa.the mitochondrial outer membrane mediates numerous interactions between the metabolic and genetic systems of mitochondria and the rest of the eukaryotic cell. we performed a proteomic study to discover novel functions of components of the mitochondrial outer membrane. proteins of highly pure outer membrane vesicles (omv) from neurospora crassa were identified by a combination of lc-ms/ms of tryptic peptide digests and gel electrophoresis of solubilized omv proteins, followed by their identificat ...200616294304
photomorphogenetic characteristics are severely affected in nucleoside diphosphate kinase-1 (ndk-1)-disrupted mutants in neurospora crassa.we previously demonstrated that the ndk-1 (nucleoside diphosphate kinase-1) point mutant, ndk-1(p72h), displays a defective phenotype in light-induced perithecial polarity in neurospora crassa. to investigate the biological function of ndk-1 in detail, we isolated two ndk-1 mutants, ndk-1(rip-1) and ndk-1(rip-2), using the riping (repeat induced point mutation) method. notably, we detected no accumulation of ndk-1(rip-1) mrna and truncated ndk-1(rip-2) protein. the ndk-1(rip) mutants exhibited a ...200616307287
bioclonedb: a database application to manage dna sequence and gene expression data.bioclonedb is a user-friendly database with a web interface to assist molecular genetics laboratories in managing a local repository of sequence information linked to dna clones. this tool is designed to assist in high-throughput sequence and gene expression projects, providing a link between both types of information. the unique feature of the application is the automation of batch sequence annotation following blast((r)) searches, which is supported by easy-to-use web interfaces. furthermore, ...200516309345
the relationship between frq-protein stability and temperature compensation in the neurospora circadian clock.temperature compensation is an important property of all biological clocks. in neurospora crassa, negative-feedback regulation on the frequency (frq) gene's transcription by the frq protein plays a central role in the organism's circadian pacemaker. earlier model calculations predicted that the stability of frq should determine the period length of neurospora's circadian rhythm as well as the rhythm's temperature compensation. here, we report experimental frq protein stabilities in frq mutants a ...200516314576
genetic and molecular analysis of phytochromes from the filamentous fungus neurospora crassa.phytochromes (phys) comprise a superfamily of red-/far-red-light-sensing proteins. whereas higher-plant phys that control numerous growth and developmental processes have been well described, the biochemical characteristics and functions of the microbial forms are largely unknown. here, we describe analyses of the expression, regulation, and activities of two phys in the filamentous fungus neurospora crassa. in addition to containing the signature n-terminal domain predicted to covalently associ ...200516339731
extraction and quantification of solutes in solidified agar culture media.a method is described for determining the concentration of certain solutes in solidified culture media. the method is based upon the finding that under specified conditions the concentration of solute in an agar gel (c(g)) is related to the concentration of solute in a centrifugally extracted gel supernatant (c(s)) by the ratio, c(g)/c(s), which is characteristic for each solute. the method avoids direct assays of the gels and instead involves assaying the supernatants from inoculated and uninoc ...197916345342
cell wall composition of neurospora crassa under conditions of copper toxicity.the mycelia of neurospora crassa grown in the presence of high concentrations of copper were blue in color, but only on a medium containing inorganic nitrate and phosphate as the nitrogen and phosphate sources, respectively. the cell wall isolate of the blue mycelia contained large amounts (12%) of copper and higher amounts of chitosan, phosphate, and amino groups, with a 42% decrease in the chitin content. although all the glucosamine of the cell wall of control cultures could be released withi ...198316346385
production and properties of extracellular endoxylanase from neurospora crassa.neurospora crassa 870 produced 14 and 0.025 u of extracellular xylanase (1,4-beta-d-xylan xylanohydrolase; ec 3.2.1.8) and beta-xylosidase (1,4-beta-xylan xylohydrolase; ec 3.2.1.37) per ml, respectively, in 4 days when commercial xylan was used as a carbon source. the effects of ph and carbon sources on xylanase production by n. crassa are discussed. two xylanases (i and ii) were purified and had pi values of 4.8 and 4.5 and molecular weights of 33,000 and 30,000. the maximum degree of hydrolys ...198416346591
genetic transformation of the biocontrol fungus gliocladium virens to benomyl resistance.methodology was developed to isolate and regenerate protoplasts from the biocontrol fungus gliocladium virens and to transform them to benomyl resistance with a neurospora crassa beta-tubulin gene. southern blots demonstrated that multiple copies of the vector integrated into the chromosomal dna of stable biotypes but not of abortive transformants. analysis of nuclear condition in vegetative and asexual structures demonstrated that no structure of g. virens is dependably uninucleate and thus pre ...199016348312
occurrence of nitrate reductase and molybdopterin in xanthomonas maltophilia.fifteen of 23 atcc strains and 2 of 9 clinical isolates of xanthomonas maltophilia, all of which grew aerobically on ammonia, but not nitrate, as a sole nitrogen source, reduced nitrate to nitrite. x. maltophilia failed to grow anaerobically on complex medium with or without nitrate, so it is considered an obligate aerobe. nitrate-reducing strains contained reduced methyl viologen nitrate reductase (mvh-nr) with specific activities ranging from 49.2 to 192 u mg of protein. strain atcc 17666 doub ...199016348378
purification of two nitrate reductases from xanthomonas maltophilia grown in aerobic cultures.xanthomonas maltophilia atcc 17666 is an obligate aerobe that accumulates nitrite when grown on nitrate. spectra of membranes from nitrate-grown cells exhibited b-type cytochrome peaks and a(615-630) indicative of d-type cytochrome but no absorption peaks corresponding to c-type cytochromes. the nitrate reductase (nr) activity was located in the membrane fraction. triton x-100-extracted reduced methyl viologen-nrs were purified on de-52, hydroxylapatite, and sephacryl s-300 columns to specific a ...199216348805
laser microsurgery permits fungal plasma membrane single-ion-channel resolution at the hyphal tip.a method for formation of high-electrical-resistance seals on the neurospora crassa plasma membrane, allowing resolution of single-ion-channel activity by patch clamp electrophysiology, is reported. laser microsurgery permits access to the hyphal apex without enzymatic cell wall digestion and loss of morphological polarity. cell wall reformation is delayed by brefeldin. this method can allow full characterization of apical plasma membrane channels, which are implicated in tip growth.199816349556
glutamine metabolism and cycling in neurospora crassa.[this corrects the article on p. toc in vol. 54.].199016350247
isolation and characterization of a sodium-dependent phosphate transporter gene in dunaliella viridis.a sodium-dependent phosphate transporter gene, dvspt1, was isolated from a cdna library using a probe derived from a subtracted cdna library of dunaliella viridis. sequencing analyses revealed a cdna sequence of 2649 bp long and encoded an open-reading frame consisting of 672 amino acids. the deduced amino acid sequence of dvspt1 exhibited 31.2% identity to that of tcpho from tetraselmis chui. hydrophobicity and secondary structure prediction revealed 11 conserved transmembrane domains similar t ...200616359638
structural analysis of the zen-4/cemklp1 motor domain and its interaction with microtubules.the centralspindlin complex is required for the assembly and maintenance of the central spindle during late anaphase and the completion of cytokinesis. it is composed of two copies each of the kinesin-like protein zen-4, a caenorhabditis elegans mklp-1 (kinesin-6 family), and the rhogap cyk-4. by using cryo-electron microscopy and helical 3d reconstruction, we are investigating the structural features of the interactions between monomeric and dimeric motor domain constructs of zen-4 and microtub ...200616361109
chromosome segment duplications in neurospora crassa and their effects on repeat-induced point mutation and meiotic silencing by unpaired dna.the size and extent of four neurospora crassa duplications, dp(ar17), dp(ibj5), dp(oy329), and dp(b362i), was determined by testing the coverage of rflp markers. the first three duplications were all > approximately 350 kb and have been shown in earlier studies to act as dominant suppressors of repeat-induced point mutation (rip) in gene-sized duplications, possibly via titration of the rip machinery. dp(b362i), which is only approximately 117 kb long, failed to suppress rip. rip suppression in ...200616361235
biogenesis and replication of small plasmid-like derivatives of the mitochondrial dna in neurospora crassa.for reasons that are not obvious, sets of related, small, plasmid-like elements appear spontaneously and become amplified in the mitochondria of some cytochrome-deficient and/or uv-sensitive mutants of neurospora crassa. these plasmid-like dnas are multimeric series of circular molecules, each consisting of a finite number of identical tandem repeats of a relatively short mtdna-derived nucleotide sequence (monomer). the plasmid-like elements that have been characterized in this study consist of ...200616386436
asexual transmission, non-suppressiveness and meiotic extinction of small plasmid-like derivatives of the mitochondrial dna in neurospora crassa.for reasons that are not obvious, sets of related plasmid-like elements that consist of short segments of dna that overlap the 5' terminal region of the mitochondrial large-subunit rrna gene sometimes appear spontaneously and become amplified in the mitochondria of some cytochrome-deficient and/or uv-sensitive mutants of neurospora crassa. these elements are transmitted efficiently through hyphal anastomoses and appear to invade the mitochondria of recipient strains, but they do not cause senesc ...200616386438
molecular cloning, expression, and characterization of the cu,zn superoxide dismutase (sod1) gene from the entomopathogenic fungus cordyceps militaris.we describe the molecular characterization of the cu,zn superoxide dismutase (sod1) gene of cordyceps militaris, which is one of the entomopathogenic fungi called a vegetable wasp and plant worm. the sod1 gene of c. militaris spans 922 bp and consisted of three introns and four exons coding for 154 amino acid residues. the deduced amino acid sequence of the c. militaris sod1 cdna showed 88% identity to claviceps purpurea sod1, 82% to neurospora crassa sod1, and 75-64% to sod1 sequences from othe ...200516389964
[development of pythium ultimum var. ultimum trow on different cultural media depending on temperature].pythium ultimum var. ultimum 175 was isolated for the first time from root system of rape. influences of two kinds of temperature (10 and 25 degrees c) and 8 different cultural media on the strain growth and development was investigated. using the growth-rate criterion the author has determined linear velocity of growth, which varied from 0.43 to 0.68 mm/h at 10 degrees c and from 0.78 to 1.11 mm/h at 25 degrees c. as to the velocity of growth the culture p. ultimum var. ultimum 175 is comparabl ...200516396111
the presence of gc-ag introns in neurospora crassa and other euascomycetes determined from analyses of complete genomes: implications for automated gene prediction.a combination of experimental and computational approaches was employed to identify introns with noncanonical gc-ag splice sites (gc-ag introns) within euascomycete genomes. evaluation of 2335 cdna-confirmed introns from neurospora crassa revealed 27 such introns (1.2%). a similar frequency (1.0%) of gc-ag introns was identified in fusarium graminearum, in which 3 of 292 cdna-confirmed introns contained gc-ag splice sites. computational analyses of the n. crassa genome using a gc-ag intron conse ...200616406724
genomics reveals traces of fungal phenylpropanoid-flavonoid metabolic pathway in the f ilamentous fungus aspergillus oryzae.fungal secondary metabolites constitute a wide variety of compounds which either play a vital role in agricultural, pharmaceutical and industrial contexts, or have devastating effects on agriculture, animal and human affairs by virtue of their toxigenicity. owing to their beneficial and deleterious characteristics, these complex compounds and the genes responsible for their synthesis have been the subjects of extensive investigation by microbiologists and pharmacologists. a majority of the funga ...200516410762
fungal heat-shock proteins in human disease.heat-shock proteins (hsps) have been identified as molecular chaperones conserved between microbes and man and grouped by their molecular mass and high degree of amino acid homology. this article reviews the major hsps of saccharomyces cerevisiae, their interactions with trehalose, the effect of fermentation and the role of the heat-shock factor. information derived from this model, as well as from neurospora crassa and achlya ambisexualis, helps in understanding the importance of hsps in the pa ...200616438680
homology effects in neurospora crassa.it has become clear in the past few years that eukaryotic organisms possess different genetic systems to counter viruses, transposons and other repeated elements such as transgenes that could otherwise accumulate in the genome. in addition to serving as a model organism for genetic, biochemical and molecular studies, neurospora crassa has proved to be a paradigm for the study of gene-silencing mechanisms. indeed, its genome can be protected from expansion of selfish nucleic acids by a variety of ...200616445744
identification and comparative analysis of the large subunit mitochondrial ribosomal proteins of neurospora crassa.the mitochondrial ribosome (mitoribosome) has highly evolved from its putative prokaryotic ancestor and varies considerably from one organism to another. to gain further insights into its structural and evolutionary characteristics, we have purified and identified individual mitochondrial ribosomal proteins of neurospora crassa by mass spectrometry and compared them with those of the budding yeast saccharomyces cerevisiae. most of the mitochondrial ribosomal proteins of the two fungi are well co ...200616451194
on the presence of a heme-binding domain homologous to cytochrome b(5) in neurospora crassa assimilatory nitrate reductase.assimilatory nitrate reductase has been purified with 55% recovery from a neurospora crassa nmr-1 nit-6 mutant, using a modification of a published procedure. it possesses one heme per 240 000 g, and subunits of mol. wt. 68 000. upon digestion with chymotrypsin, a heme-binding domain was isolated by gel filtration; its visible spectrum was highly similar to that of cytochrome b(5). on sds gels, the fraction showed two heme-containing bands of 10 000 and 12 5000 daltons; their amino acid composit ...198316453480
mannosyltransferase is required for cell wall biosynthesis, morphology and control of asexual development in neurospora crassa.two neurospora mutants with a phenotype that includes a tight colonial growth pattern, an inability to form conidia and an inability to form protoperithecia have been isolated and characterized. the relevant mutations were mapped to the same locus on the sequenced neurospora genome. the mutations responsible for the mutant phenotype then were identified by examining likely candidate genes from the mutant genomes at the mapped locus with pcr amplification and a sequencing assay. the results demon ...200616457356
cell-wall-bound metal ions are not taken up in neurospora crassa.to establish the relevance of the cell wall in metal ion transport, cobalt uptake was examined in neurospora crassa. cobalt taken up was largely surface bound (>90%), resulting in a release of calcium and magnesium. surface-bound cobalt could not enter intracellular locations upon further incubation of mycelia in a metal-free medium. saturation of the surface with one metal augured subsequent dose-dependent entry of a different metal into intracellular locations. in comparison with the cobalt-re ...200516462860
construction of pdestr, a gateway vector for gene disruption in filamentous fungi.we have constructed pdestr, a destination vector of gateway system especially for gene targeting and disruption in filamentous fungi. the vector was constructed by removing the multicloning site of pgem-t easy vector, and inserting hygromycin phosphotransferase gene construct from pcb1004, and a gateway vector conversion cassette. in order to construct a dna for gene disruption, only an inverse-polymerase chain reaction (pcr) amplification of the restricted, target sequence is needed. after the ...200616479358
microarray and real-time pcr analyses reveal mating type-dependent gene expression in a homothallic fungus.sordaria macrospora is a homothallic ascomycete which is able to form fertile fruiting bodies without a mating partner. to analyze the molecular basis of homothallism and the role of mating products during fruiting body development, we have deleted the mating type gene smta-1 encoding a high-mobility group domain (hmg) protein. the deltasmta-1 deletion strain is morphologically wild type during vegetative growth, but it is unable to produce perithecia or ascospores. to identify genes expressed u ...200616482473
[cloning and characterization of glyceraldehyde-3-phosphate dehydrogenase gene from chaetomium globosum].the amino acid sequence of glyceraldehyde 3-phosphate dehydrogenase (gapdh) gene from neurospora crassa (xp-327967) and colletotrichum lindemuthianu (p35143) were subjected to local tblastn searching against the ests local datebase of chaetomium globosum. the full length cdna sequence of 1240bp encoding gapdh gene with an open reading frame of 1014bp and encoding 337 amino acids was obtained. the protein molecular weight was 36.1kd. the dna sequence of gapdh gene was obtained through pcr amplifi ...200516496697
deletion variants of neurospora mitochondrial porin: electrophysiological and spectroscopic analysis.mitochondrial porins are predicted to traverse the outer membrane as a series of beta-strands, but the precise structure of the resulting beta-barrel has remained elusive. toward determining the positions of the membrane-spanning segments, a series of small deletions was introduced into several of the predicted beta-strands of the neurospora crassa porin. overall, three classes of porin variants were identified: i), those producing large, stable pores, indicating deletions likely outside of beta ...200616500966
nonribosomal peptide synthesis in schizosaccharomyces pombe and the architectures of ferrichrome-type siderophore synthetases in fungi.a nonribosomal peptide synthetase (nrps) in schizosaccharomyces pombe, which possesses an unusual structure incorporating three adenylation domains, six thiolation domains and six condensation domains, has been shown to produce the cyclohexapeptide siderophore ferrichrome. one of the adenylation domains is truncated and contains a distorted key motif. substrate-binding specificities of the remaining two domains were assigned by molecular modelling to glycine and to n-acetyl-n-hydroxy-l-ornithine ...200616502473
depletion of aspergillus nidulans cota causes a severe polarity defect which is not suppressed by the nuclear migration mutation nuda2.the aspergillus nidulans homologue of neurospora crassa cot-1, cota, encoding a member of the ndr protein kinase family, has been cloned and expressed under the control of the conditional alca promoter. depletion of cota by repression of the alca promoter led to a severe growth defect accompanied by loss of polarity. germlings show greatly enlarged volume of the spores and hyphae, accompanied by an increase in number of nuclei per compartment, though the nucleus/volume ratio is not significantly ...200616506053
[analysis of the secreted proteins encoded by genes in genome of filamental fungus (neurospora crassa)].the completed genome sequence of filamental fungus neurospora crassa has completed and 10,082 open reading frames (orfs) were predicted in whole genome. among them, 437 proteins were secreted ones based on the combination of prediction algorithms signalp v3.0 and psort, transmembrane domains prediction algorithms tmhmm v2.0 and thumbup, potential gpi-anchor sites prediction algorithm big-pi predictor and the subcellular protein location prediction algorithm targetp v1.01. the minimum orf length ...200616520317
pheromones are essential for male fertility and sufficient to direct chemotropic polarized growth of trichogynes during mating in neurospora crassa.neurospora crassa is a self-sterile filamentous fungus with two mating types, mat a and mat a. its mating involves chemotropic polarized growth of female-specific hyphae (trichogynes) toward male cells of the opposite mating type in a process involving pheromones and receptors. mat a cells express the ccg-4 pheromone and the pre-1 receptor, while mat a strains produce mrna for the pheromone mfa-1 and the pre-2 receptor; mfa-1 and ccg-4 are the predicted ligands for pre-1 and pre-2, respectively. ...200616524909
mutational analysis of the glycosylphosphatidylinositol (gpi) anchor pathway demonstrates that gpi-anchored proteins are required for cell wall biogenesis and normal hyphal growth in neurospora crassa.using mutational and proteomic approaches, we have demonstrated the importance of the glycosylphosphatidylinositol (gpi) anchor pathway for cell wall synthesis and integrity and for the overall morphology of the filamentous fungus neurospora crassa. mutants affected in the gpig-1, gpip-1, gpip-2, gpip-3, and gpit-1 genes, which encode components of the n. crassa gpi anchor biosynthetic pathway, have been characterized. gpi anchor mutants exhibit colonial morphologies, significantly reduced rates ...200616524913
evidence of rip (repeat-induced point mutation) in transposase sequences of aspergillus oryzae.a dna methyl-binding column was used to isolate genomic fragments enriched for dna-methylation from aspergillus parasiticus. one of the isolated sequences presented 67% identity at the protein level with the transposase from the transposable element tan1 of aspergillus niger var. awamori, and was found to be present in at least 20 copies in the aspergillus oryzae database. analysis of four copies showed evidence of c:g to t:a transitions in at least 98.2% of the mutations found over a 1,032-1,18 ...200616531081
close correlation between heat shock response and cytotoxicity in neurospora crassa treated with aliphatic alcohols and phenols.in neurospora crassa the aliphatic alcohols methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, isobutanol, ethylene glycol, glycerol, and allyl alcohol and the phenolic compounds phenol, hydroquinone, resorcinol, pyrogallol, phloroglucinol, sodium salicylate, and acetylsalicylic acid were analyzed with respect to their capacities to induce heat shock proteins (hsp) and to inhibit protein synthesis. both the alcohols and phenols showed the greatest levels of hsp induction at concent ...199516534981
primer sets developed to amplify conserved genes from filamentous ascomycetes are useful in differentiating fusarium species associated with conifers.we examined the usefulness of primer sets designed to amplify introns within conserved genes in filamentous ascomycetes to differentiate 35 isolates representing six different species of fusarium commonly found in association with conifer seedlings. we analyzed restriction fragment length polymorphisms (rflp) in five amplified pcr products from each fusarium isolate. the primers used in this study were constructed on the basis of sequence information from the h3, h4, and (beta)-tubulin genes in ...199516534991
circadian clock genes frequency and white collar-1 are not essential for entrainment to temperature cycles in neurospora crassa.the fungus neurospora crassa is a model system for investigating the mechanism of circadian rhythmicity, and the core of its circadian oscillator is thought to be a transcription/translation feedback loop involving the products of the frq (frequency), wc-1 (white-collar-1) and wc-2 (white-collar-2) genes. several reports of rhythmicity in frq and wc null mutants have raised questions about how central the frq/wc loop is to the circadian system of neurospora. several research groups have attempte ...200616537415
nonallelic interactions between het-c and a polymorphic locus, pin-c, are essential for nonself recognition and programmed cell death in neurospora crassa.nonself recognition in filamentous fungi is conferred by genetic differences at het (heterokaryon incompatibility) loci. when individuals that differ in het specificity undergo hyphal fusion, the heterokaryon undergoes a programmed cell death reaction or is highly unstable. in neurospora crassa, three allelic specificities at the het-c locus are conferred by a highly polymorphic domain. this domain shows trans-species polymorphisms indicative of balancing selection, consistent with the role of h ...200616554411
location of aryl sulfatase in conidia and young mycelia of neurospora crassa.aryl sulfatase (arylsulfate sulfohydrolase, ec 3.1.6.1) was found to have multiple locations in neurospora conidia. some enzyme activity remained in the supernatant when a spore suspension was centrifuged or filtered. part of the cell-bound activity could be detected by adding the assay ingredients to a suspension of intact spores (patent enzyme), and additional activity was only detectable when the spores were first treated to destroy their permeability barriers (cryptic enzyme). such treatment ...197016559101
chemical activation of ascospore germination in neurospora crassa. 194816561462
pigment production by neurospora crassa in the presence of para-amino-benzoic acid. 194816561489
altered responses of neurospora crassa to inhibiting concentrations of indole. 194916561804
amino acid interactions in neurospora crassa.soboren, josephine (university of california, los angeles), and joseph f. nyc. amino acid interactions in neurospora crassa. j. bacteriol. 82:20-25. 1961.-a systematic study of the effects of the naturally occurring amino acids on the growth of a wild-type strain of neurospora crassa focused attention upon l-tryptophan, which exhibits a strong growth inhibitory effect. further investigation disclosed that other tryptophan metabolites, anthranilic acid, indole, kynurenine, and 3-hydroxykynurenine ...196116561913
inhibition of growth, synthesis, and permeability in neurospora crassa by phenethyl alcohol.lester, gabriel (reed college, portland, ore.). inhibition of growth, synthesis, and permeability in neurospora crassa by phenethyl alcohol. j. bacteriol. 90: 29-37. 1965.-inhibition of the growth of neurospora crassa in still culture was detected at 0.05% and was complete at a level of 0.2% phenethyl alcohol (pea). benzyl alcohol was less inhibitory, and 3-phenyl-1-propanol and phenol were more inhibitory, than pea; benzylamine and phenethylamine were less inhibitory than the analogous hydroxyl ...196516562036
developmental expression of two forms of arginase in neurospora crassa.n. crassa has two forms of arginase. the physiological role of multiple arginases is not understood. the two forms were shown to be differentially expressed from a single locus (aga) and both proteins are localized to the cytoplasm. the 36-kda protein was expressed in minimal and arginine supplemented medium, whereas the 41-kda form was detected only in the presence of arginine. in this study we examined developmental expression of the two arginase transcripts and proteins in conidia and during ...200616574329
the genetic control of adenylosuccinase in neurospora crassa. 195716578469
a case of "maternal" inheritance in neurospora crassa. 195216589122
the accumulation of alpha-phenylglycine by mutants of neurospora crassa stimulated by phenylalanine and tyrosine. 195216589129
pseudo-wild types in neurospora crassa. 195216589148
mendelian and non-mendelian factors affecting the cytochrome system in neurospora crassa. 195316589312
a partial map of linkage group d in neurospora crassa. 195416589501
on the glutamate-proline-ornithine interrelation in neurospora crassa. 195416589539
the q locus of neurospora crassa. 195616589847
heterocaryosis and protoplasmic incompatibility in neurospora crassa. 195616589916
synergism of two cytoplasmically inherited mutants in neurospora crassa. 195616589942
evidence from tetrad analysis for both normal and aberrant recombination between allelic mutants in neurospora crassa. 195816590210
production, reproduction, and reversion of protoplast-like structures in the osmotic strain of neurospora crassa. 195816590257
the biosynthesis of isoleucine and valine in neurospora crassa. 195816590304
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