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cobalt-resistance in wall-less mutant (fz; sg; os-1) of neurospora crassa.a cobalt-resistant wall-less mutant (slime) of neurospora crassa was obtained by repeated sub-culturing of the sensitive wall-less mutant (w-sl) on agar medium containing toxic concentrations of cobalt. resistance was stable on culturing cor-sl on cobalt-free medium up to 15 weekly subcultures. cor-sl is 10-fold more resistant to cobalt when compared to w-sl. it is also cross-resistant to cu (10-fold) and ni (3-fold). cobalt accumulated by cor-sl during growth and in short-term uptake experiment ...200312779238
hop, an active mutator-like element in the genome of the fungus fusarium oxysporum.a new type of active dna transposon has been identified in the genome of fusarium oxysporum by its transposition into the niad target gene. two insertions within the final exon, in opposite orientations at the same nucleotide site, have been characterized. these elements, called hop, are 3,299 bp long, with perfect terminal inverted repeats (tirs) of 99 bp. the sequencing of genomic copies reveals a 9-bp target site duplication and no apparent sequence specificity at the insertion sites. the seq ...200312777515
biosynthesis of pyridoxine in saccharomyces cerevisiae--origin of the pyridoxine nitrogen atom differs under anaerobic and aerobic conditions.the amide nitrogen atom of glutamine is incorporated into pyridoxine in four eukaryotes (i.e., emericella nidulans, mucor racemosus, neurospora crassa and saccharomyces cerevisiae) and two prokaryotes (i.e., staphylococcus aureus and bacillus subtilis). however, in the prokaryotes pseudomonas putida, enterobacter aerogenes and escherichia coli, it is the nitrogen atom of glutamate that is incorporated into pyridoxine (j nutr sci vitaminol (2000) 46, 55-57). as these results were from experiments ...200212775110
production and characterization of the talaromyces stipitatus feruloyl esterase faec in pichia pastoris: identification of the nucleophilic serine.feruloyl esterases constitute an interesting group of enzymes that have the potential for use over a broad range of applications in the agri-food industries. we report the over-expression and characterization of a novel feruloyl esterase exhibiting broad substrate specificity from talaromyces stipitatus (faec) in pichia pastoris. using various gene constructions, we have investigated the use of alternative signal peptides to produce an authentic feruloyl esterase featuring the n-terminal sequenc ...200312767807
the role of adaptor protein ste50-dependent regulation of the mapkkk ste11 in multiple signalling pathways of yeast.in saccharomyces cerevisiae, ste50 functions in cell signalling between the activated g protein and the mitogen-activated protein kinase (mapk) kinase kinase (mapkkk) ste11. scste50 is an essential component of three mapk-mediated signalling pathways, which control the mating response, invasive/filamentous growth and osmotolerance (hog pathway), respectively. scste50 signalling may also contribute to cell wall integrity in vegetative cells. the protein contains a sterile alpha motif (sam) and a ...200312764668
protonation of the neurospora crassa plasma membrane h+-atpase as a function of ph monitored by atr-ftir. 200312763847
molecular genetics of fungal siderophore biosynthesis and uptake: the role of siderophores in iron uptake and storage.to acquire iron, all species have to overcome the problems of iron insolubility and toxicity. in response to low iron availability in the environment, most fungi excrete ferric iron-specific chelators--siderophores--to mobilize this metal. siderophore-bound iron is subsequently utilized via the reductive iron assimilatory system or uptake of the siderophore-iron complex. furthermore, most fungi possess intracellular siderophores as iron storage compounds. molecular analysis of siderophore biosyn ...200312759789
the 9.8 kda subunit of complex i, related to bacterial na(+)-translocating nadh dehydrogenases, is required for enzyme assembly and function in neurospora crassa.a nuclear gene encoding a 9.8 kda subunit of complex i, the homologue of mammalian mwfe protein, was identified in the genome of neurospora crassa. the gene was cloned and inactivated in vivo by the generation of repeat-induced point mutations. fungal mutant strains lacking the 9.8 kda polypeptide were subsequently isolated. analyses of mitochondrial proteins from mutant nuo9.8 indicate that the membrane and peripheral arms of complex i fail to assemble. respiration of mutant mitochondria on mat ...200312758076
diversification of exogenous genes in vivo in neurospora.we have adapted the meiotic recombination hotspot cog of neurospora crassa for shuffling exogenous dna, providing a means of generating novel genes in situ from sequences introduced into chromosomes. genes to be diversified are inserted between the his-3 locus and cog. diversification crosses are heterozygous both for alleles of the exogenous dna and for auxotrophic alleles of his-3. progeny selected for ability to grow without histidine supplementation are enriched for exchange events within th ...200312756504
analysis of loss of pathogenicity mutants reveals that repeat-induced point mutations can occur in the dothideomycete leptosphaeria maculans.restriction enzyme mediated insertional mutagenesis using a plasmid, pucatph, that confers hygromycin resistance, generated loss-of-pathogenicity mutants of leptosphaeria maculans, the fungus that causes blackleg disease of brassica napus. of 516 l. maculans transformants analysed, 12 were pathogenicity mutants. when eight of these mutants were crossed to an isolate that attacks b. napus, cosegregation of pucatph sequences and loss of pathogenicity was not observed, suggesting that these mutatio ...200312742061
a complete inventory of fungal kinesins in representative filamentous ascomycetes.complete inventories of kinesins from three pathogenic filamentous ascomycetes, botryotinia fuckeliana, cochliobolus heterostrophus, and gibberella moniliformis, are described. these protein sequences were compared with those of the filamentous saprophyte, neurospora crassa and the two yeasts saccharomyces cerevisiae and schizosaccharomyces pombe. data mining and phylogenetic analysis of the motor domain yielded a constant set of 10 kinesins in the filamentous fungal species, compared with a sma ...200312742059
deletion of mdmb impairs mitochondrial distribution and morphology in aspergillus nidulans.mitochondria form a dynamic network of interconnected tubes in the cells of saccharomyces cerevisiae or filamentous fungi such as aspergillus nidulans, neurospora crassa, or podospora anserina. the dynamics depends on the separation of mitochondrial fragments, their movement throughout the cell, and their subsequent fusion with the other parts of the organelle. interestingly, the microtubule network is required for the distribution in n. crassa and s. pombe, while s. cerevisiae and a. nidulans a ...200312740872
the yeast isn1 (yor155c) gene encodes a new type of imp-specific 5'-nucleotidase.the purine salvage enzyme inosine 5'-monophosphate (imp)-specific 5'-nucleotidase catalyzes degradation of imp to inosine. although this enzymatic activity has been purified and characterized in saccharomyces cerevisiae, the gene encoding imp 5'-nucleotidase had not been identified.200312735798
neurospora crassa fks protein binds to the (1,3)beta-glucan synthase substrate, udp-glucose.the essential fungal cell-wall polymer (1,3)beta-glucan is synthesized by the enzyme (1,3)beta-glucan synthase. this enzyme, which is the target of the echinocandin and pneumocandin families of fungicidal antibiotics, is a complex composed of at least two proteins, rho1p and fks1p. homologs of the yeast fks1 gene have been discovered in numerous fungi, and existing evidence points to, but has not yet proved, fks1p being the catalytic subunit of (1,3)beta-glucan synthase. we have purified (1,3)be ...200312732946
synthesis and conformational characterization of peptides related to the neck domain of a fungal kinesin.the y362k mutation in the neck domain of conventional kinesin from neurospora crassa provokes a significant reduction of the rate of movement along microtubules. since the alpha-helical coiled-coil structure of the neck region is implicated in the mechanism of the processive movement of kinesins, a series of peptides related to the heptad region 338-379 of the wild-type and the variant fungal kinesinswere synthesized as monomers and as n-terminal disulfide dimers, crosslinked to favour self-asso ...200312725241
rhythmic binding of a white collar-containing complex to the frequency promoter is inhibited by frequency.the biological clock of neurospora crassa includes interconnected transcriptional and translational feedback loops that cause both the transcript and protein encoded by the frequency gene (frq) to undergo the robust daily oscillations in abundance, which are essential for clock function. to understand better the mechanism generating rhythmic frq transcript, reporter constructs were used to show that the oscillation in frq message is transcriptionally regulated, and a single cis-acting element in ...200312714686
enzymes with extra talents: moonlighting functions and catalytic promiscuity.recent studies of moonlighting functions and catalytic promiscuity provide insights into the structural and mechanistic bases of these phenomena. moonlighting proteins that are highlighted include gephyrin, the neurospora crassa tyrosyl trna synthetase, phosphoglucose isomerase, and cytochrome c. new insights into catalytic promiscuity are provided by studies of aminoglycoside kinase (3') type iiia, tetrachlorohydroquinone dehalogenase, and aldolase antibody 38c2.200312714060
the methylated component of the neurospora crassa genome.cytosine methylation is common, but not ubiquitous, in eukaryotes. mammals and the fungus neurospora crassa have about 2-3% of cytosines methylated. in mammals, methylation is almost exclusively in the under-represented cpg dinucleotides, and most cpgs are methylated whereas in neurospora, methylation is not preferentially in cpg dinucleotides and the bulk of the genome is unmethylated. dna methylation is essential in mammals but is dispensable in neurospora, making this simple eukaryote a favou ...200312712205
the genome sequence of the filamentous fungus neurospora crassa.neurospora crassa is a central organism in the history of twentieth-century genetics, biochemistry and molecular biology. here, we report a high-quality draft sequence of the n. crassa genome. the approximately 40-megabase genome encodes about 10,000 protein-coding genes--more than twice as many as in the fission yeast schizosaccharomyces pombe and only about 25% fewer than in the fruitfly drosophila melanogaster. analysis of the gene set yields insights into unexpected aspects of neurospora bio ...200312712197
promoter analysis of the avirulence gene avr9 of the fungal tomato pathogen cladosporium fulvum in the model filamentous fungus aspergillus nidulans.the promoter of avirulence gene avr9 of the fungal tomato pathogen cladosporium fulvum contains 12 sequences within a region of 0.6 kb that are reminiscent of the binding sequences of the gata-type regulator involved in nitrogen utilisation of the filamentous fungi aspergillus nidulans and neurospora crassa. mutational analysis of this 0.6-kb promoter region, fused to the beta-glucuronidase reporter gene, revealed that two regions, each containing two tagata boxes in inverted orientation and ove ...200312695849
construction of strains for rapid homokaryon purification after integration of constructs at the histidine-3 ( his-3) locus of neurospora crassa.we report the construction of histidine-3 (his-3) strains of neurospora crassa containing the hygromycin b phosphotransferase gene of escherichia coli (hph(+)) fused in-frame to the herpes simplex virus thymidine kinase gene (tk(+); lupton et al. 1991), integrated at the his-3 locus. we also report the construction of two ampicillin-resistant and two kanamycin-resistant his-3 gene-replacement vector plasmids. the combined use of these strains and plasmids for his-3-targeted gene integration allo ...200312684841
histone h1 is required for proper regulation of pyruvate decarboxylase gene expression in neurospora crassa.we show that neurospora crassa has a single histone h1 gene, hh1, which encodes a typical linker histone with highly basic n- and c-terminal tails and a central globular domain. a green fluorescent protein-tagged histone h1 chimeric protein was localized exclusively to nuclei. mutation of hh1 by repeat-induced point mutation (rip) did not result in detectable defects in morphology, dna methylation, mutagen sensitivity, dna repair, fertility, rip, chromosome pairing, or chromosome segregation. ne ...200312684383
a circadian oscillator in aspergillus spp. regulates daily development and gene expression.we have established the presence of a circadian clock in aspergillus flavus and aspergillus nidulans by morphological and molecular assays, respectively. in a. flavus, the clock regulates an easily assayable rhythm in the development of sclerotia, which are large survival structures produced by many fungi. this developmental rhythm exhibits all of the principal clock properties. the rhythm is maintained in constant environmental conditions with a period of 33 h at 30 degrees c, it can be entrain ...200312684372
circadian and light-induced expression of luciferase in neurospora crassa.we have constructed a plasmid vector for expressing firefly luciferase in neurospora crassa under control of the light- and clock-regulated ccg-2 (eas) promoter. the sequence of the luciferase gene in the vector has been modified to reflect the n. crassa codon bias. both light-induced activity and circadian activity are demonstrated. expression of luciferase in strains carrying mutant frequency alleles shows appropriate period length alterations. these data demonstrate that luciferase is a sensi ...200312684022
trimethylated lysine 9 of histone h3 is a mark for dna methylation in neurospora crassa.besides serving to package nuclear dna, histones carry information in the form of a diverse array of post-translational modifications. methylation of histones h3 and h4 has been implicated in long-term epigenetic 'memory'. dimethylation or trimethylation of lys4 of histone h3 (h3 lys4) has been found in expressible euchromatin of yeasts and mammals. in contrast, methylation of lys9 of histone h3 (h3 lys9) has been implicated in establishing and maintaining the largely quiescent heterochromatin o ...200312679815
purification and characterization of the complex i from the respiratory chain of rhodothermus marinus.the rotenone sensitive nadh:menaquinone oxidoreductase (ndh-i or complex i) from the thermohalophilic bacterium rhodothermus marinus has been purified and characterized. three of its subunits react with antibodies against 78, 51, and 21.3c kda subunits of neurospora crassa complex i. the optimum conditions for nadh dehydrogenase activity are 50 degrees c and ph 8.1, and the enzyme presents a km of 9 microm for nadh. the enzyme also displays nadh:quinone oxidoreductase activity with two menaquino ...200212678433
what's in the genome of a filamentous fungus? analysis of the neurospora genome sequence.the german neurospora genome project has assembled sequences from ordered cosmid and bac clones of linkage groups ii and v of the genome of neurospora crassa in 13 and 12 contigs, respectively. including additional sequences located on other linkage groups a total of 12 mb were subjected to a manual gene extraction and annotation process. the genome comprises a small number of repetitive elements, a low degree of segmental duplications and very few paralogous genes. the analysis of the 3218 iden ...200312655011
analysis of interactions between domains of a small heat shock protein, hsp30 of neurospora crassa.the alpha-crystallin-related, small heat shock proteins (shsps), despite their overall variability in sequence, have discrete regions of conserved sequence that are involved in structural organization, as well as nonconserved regions that may perform similar roles in each protein. recent x-ray diffraction analyses of an archeal and a plant shsp have revealed both similarities and differences in how they are organized, suggesting that there is variability, particularly in the oligomeric organizat ...200212653482
primary structure of the histone 2b gene in the white root rot fungus, rosellinia necatrix.the nucleotide sequence of the histone 2b (h2b) gene in the white root rot fungus, rosellinia necatrix, was determined. the gene has two introns in the coding region at positions conserved in the neurospora crassa and aspergillus nidulans h2b genes, but the third intron present in the h2b gene from n. crassa and a. nidulans is absent in the r. necatrix h2b gene. the amino acid sequence of the coding region of the r. necatrix gene resembled that of n. crassa and a. nidulans. therefore, the third ...200212652911
a hex-1 crystal lattice required for woronin body function in neurospora crassa.the woronin body is a dense-core vesicle specific to filamentous ascomycetes (euascomycetes), where it functions to seal the septal pore in response to cellular damage. the hex-1 protein self-assembles to form this solid core of the vesicle. here, we solve the crystal structure of hex-1 at 1.8 a, which provides the structural basis of its self-assembly. the structure reveals the existence of three intermolecular interfaces that promote the formation of a three-dimensional protein lattice. consis ...200312640443
synthesis of signals for de novo dna methylation in neurospora crassa.most 5-methylcytosine in neurospora crassa occurs in a:t-rich sequences high in tpa dinucleotides, hallmarks of repeat-induced point mutation. to investigate how such sequences induce methylation, we developed a sensitive in vivo system. tests of various 25- to 100-bp synthetic dna sequences revealed that both t and a residues were required on a given strand to induce appreciable methylation. segments composed of (taaa)(n) or (ttaa)(n) were the most potent signals; 25-mers induced robust methyla ...200312640122
the mapkk kinase stec regulates conidiophore morphology and is essential for heterokaryon formation and sexual development in the homothallic fungus aspergillus nidulans.environmental signals can be transduced into intracellular responses by the action of map kinase cascades. sequential phosphorylation results in the transient activation of a map kinase, which in turn activates certain transcription factors and thus a set of pathway-specific genes. many steps in this cascade are conserved, and homologues have been discovered from yeast to human. we have characterized the mapkk kinase, stec, a homologue of saccharomyces cerevisiae ste11, in the filamentous fungus ...200312622813
on the independence of barrage formation and heterokaryon incompatibility in neurospora crassa.a barrage is a line or zone of demarcation that may develop at the interface where genetically different fungi meet. barrage formation represents a type of nonself recognition that has often been attributed to the heterokaryon incompatibility system, which limits the co-occurrence of genetically different nuclei in the same cytoplasm during the asexual phase of the life cycle. while the genetic basis of the heterokaryon incompatibility system is well characterized in neurospora crassa, barrage f ...200312620257
analysis of expressed sequence tags from gibberella zeae (anamorph fusarium graminearum).gibberella zeae is a broad host range pathogen that infects many crop plants, including wheat and barley, and causes head blight and rot diseases throughout the world. to better understand fungal development and pathogenicity, we have generated 7996 ests from three cdna libraries. two libraries were generated from carbon-(c-) and nitrogen- (n-) starved mycelia and one library was generated from cultures of maturing perithecia (p). in other fungal pathogens, starvation conditions have been shown ...200312620255
regulation of expression of the rhizopus oryzae uricase and urease enzymes.the regulation of intracellular urease and uricase activities was examined in rhizopus oryzae. urease activity (2.4 u/mg protein) was present in r. oryzae mycelium grown in minimal medium containing nh4ci as sole nitrogen source. this activity increased threefold under nitrogen derepression conditions, but no induction by urea was detected. control of urease activity in r. oryzae differs from that found in neurospora crassa but resembles the situation in aspergillus nidulans. no uricase activity ...200212619824
putative homologs of ssk22 mapkk kinase and pbs2 mapk kinase of saccharomyces cerevisiae encoded by os-4 and os-5 genes for osmotic sensitivity and fungicide resistance in neurospora crassa.we cloned and characterized neurospora ncssk22 and ncpbs2 genes, similar to yeast ssk22 mitogen-activated protein (map) kinase kinase kinase and the pbs2 map kinase kinase genes, respectively. disruptants of the ncssk22 gene were sensitive to osmotic stress and resistant to iprodione and fludioxonil. their phenotypes were similar to those of osmotic-sensitive (os) mutants os-1, os-2, os-4, and os-5. the os-4 mutant strain transformed with the wild-type ncssk22 gene grew on a medium containing 4% ...200312619694
primary structure of cytochrome c gene from the white root rot fungus rosellinia necatrix.the nucleotide sequence of the cytochrome c (cytc) gene of the white root rot fungus rosellinia necatrix was analyzed. the structure of this gene, which had three introns in the coding region, was similar to that of aspergillus nidulans. the second intron of the r. necatrix cytc gene was not present in neurospora crassa or fusarium oxysporum. however, the amino acid sequence of r. necatrix was most similar to that of neurospora crassa. thus, it seemed that the second intron of the r. necatrix cy ...200312619691
inhibition of dna methylation and reactivation of silenced genes by zebularine.gene silencing by abnormal methylation of promoter regions of regulatory genes is commonly associated with cancer. silenced tumor suppressor genes are obvious targets for reactivation by methylation inhibitors such as 5-azacytidine (5-aza-cr) and 5-aza-2'-deoxycytidine (5-aza-cdr). however, both compounds are chemically unstable and toxic and neither can be given orally. we characterized a new demethylating agent, zebularine [1-(beta-d-ribofuranosyl)-1,2-dihydropyrimidin-2-one], which is a chemi ...200312618505
amplification of hmg-coa reductase production enhances carotenoid accumulation in neurospora crassa.neurospora crassa, a filamentous fungus, naturally produces the carotenoids lycopene and neurosporaxanthin. to increase the carbon flux through the carotenoid biosynthetic pathway, the 1658-bp region of the hmg1 gene encoding the catalytic domain (chmg1) of 3-hydroxy-3-methylglutaryl coenzyme a (hmg-coa) reductase of saccharomyces cerevisiae was expressed in n. crassa under control of the strong, constitutive glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter and the inducible alcohol dehyd ...200212616689
single fungal kinesin motor molecules move processively along microtubules.conventional kinesins are two-headed molecular motors that move as single molecules micrometer-long distances on microtubules by using energy derived from atp hydrolysis. the presence of two heads is a prerequisite for this processive motility, but other interacting domains, like the neck and k-loop, influence the processivity and are implicated in allowing some single-headed kinesins to move processively. neurospora kinesin (nkin) is a phylogenetically distant, dimeric kinesin from neurospora c ...200312609885
role for antisense rna in regulating circadian clock function in neurospora crassa.the prevalence of antisense rna in eukaryotes is not known and only a few naturally occurring antisense transcripts have been assigned a function. however, the recent identification of a large number of putative antisense transcripts strengthens the view that antisense rnas might affect a wider variety of processes than previously thought. here we show that in the model organism neurospora crassa entrainment of the circadian clock, which is critical for the correct temporal expression of genes a ...200312607002
the n-acetylglutamate synthase/n-acetylglutamate kinase metabolon of saccharomyces cerevisiae allows co-ordinated feedback regulation of the first two steps in arginine biosynthesis.in saccharomyces cerevisiae, which uses the nonlinear pathway of arginine biosynthesis, the first two enzymes, n-acetylglutamate synthase (nags) and n-acetylglutamate kinase (nagk), are controlled by feedback inhibition. we have previously shown that nags and nagk associate in a complex, essential to synthase activity and protein level [abadjieva, a., pauwels, k., hilven, p. & crabeel, m. (2001) j. biol. chem.276, 42869-42880]. the nagks of ascomycetes possess, in addition to the catalytic domai ...200312603335
cloning and sequence analysis of a mitochondrial gene cluster encoding cytochrome c oxidase subunit iii from trichoderma pseudokoningii.a mitochondrial gene cluster encoding cytochrome c oxidase subunit iii (cox3), an orf (called orf250) similar to nadh dehydrogenase subunit vi (nd6), ten trna molecules, partial rrna small subunit and rrna large subunit from trichoderma pseudokoningii s38 was cloned and sequenced. these genes are tandemly clustered on the mitochondrial genome of trichoderma pseudokoningii s38. phylogenetic analysis showed that cytochrome c oxidase subunits iii exhibited high degree of similarity to sequences fro ...200212592707
role of the two type ii myosins, myo2 and myp2, in cytokinetic actomyosin ring formation and function in fission yeast.the formation and contraction of a cytokinetic actomyosin ring (car) is essential for the execution of cytokinesis in fission yeast. unlike most organisms in which its composition has been investigated, the fission yeast car contains two type ii myosins encoded by the genes myo2(+) and myp2(+). myo2(+) is an essential gene whilst myp2(+) is dispensable under normal growth conditions. myo2 is hence the major contractile protein of the car whilst myp2 plays a more subtle and, as yet, incompletely ...200312589679
the qde-3 homologue recq-2 co-operates with qde-3 in dna repair in neurospora crassa.the post-transcriptional gene silencing mechanism in neurospora crassa, called quelling, was shown to involve the products of three genes termed quelling-defective. a homologue to the qde-3 gene encoding a putative recq-type dna helicase was isolated and was named recq-2. characterisation of the recq-2 gene has revealed that it is not involved in quelling, but may co-operate with the qde-3 gene product in a pathway that repairs damage to dna caused by the chemical mutagens methyl methanesulfonat ...200312589473
cooperative action of the nit2 and nit4 transcription factors upon gene expression in neurospora crassa.in neurospora crassa, the nit-3 gene, which encodes nitrate reductase, an enzyme required for the utilization of inorganic nitrate, is subject to a high degree of genetic and metabolic regulation as a member of the nitrogen control circuit. the nit-3 gene promoter contains binding sites for a globally acting protein nit2 and a pathway-specific protein nit4. expression of the nit-3 gene absolutely requires both the nit2 and nit4 transcription factors and only occurs under conditions of nitrogen s ...200312589465
pex7p and pex20p of neurospora crassa function together in pts2-dependent protein import into peroxisomes.recruiting matrix proteins with a peroxisomal targeting signal type 2 (pts2) to the peroxisomal membrane requires species-specific factors. in saccharomyces cerevisiae, the pts2 receptor pex7p acts in concert with the redundant pex18p/pex21p, whereas in yarrowia lipolytica, pex20p might unite the function of both s. cerevisiae peroxins. herein, the genome of the filamentous fungus neurospora crassa was analyzed for peroxin-encoding genes. we identified a set of 18 peroxins that resembles that of ...200312589072
roles for white collar-1 in circadian and general photoperception in neurospora crassa.the transcription factors white collar-1 (wc-1) and white collar-2 (wc-2) interact to form a heterodimeric complex (wcc) that is essential for most of the light-mediated processes in neurospora crassa. wcc also plays a distinct non-light-related role as the transcriptional activator in the frequency (frq)/wcc feedback loop that is central to the n. crassa circadian system. although an activator role was expected for wc-1, unanticipated phenotypes resulting from some wc-1 alleles prompted a close ...200312586700
single-strand-specific nucleases.single-strand-specific nucleases are multifunctional enzymes and widespread in distribution. their ability to act selectively on single-stranded nucleic acids and single-stranded regions in double-stranded nucleic acids has led to their extensive application as probes for the structural determination of nucleic acids. intracellularly, they have been implicated in recombination, repair and replication, whereas extracellular enzymes have a role in nutrition. although more than 30 single-strand-spe ...200312586391
tok homologue in neurospora crassa: first cloning and functional characterization of an ion channel in a filamentous fungus.in contrast to animal and plant cells, very little is known of ion channel function in fungal physiology. the life cycle of most fungi depends on the "filamentous" polarized growth of hyphal cells; however, no ion channels have been cloned from filamentous fungi and comparatively few preliminary recordings of ion channel activity have been made. in an attempt to gain an insight into the role of ion channels in fungal hyphal physiology, a homolog of the yeast k(+) channel (sctok1) was cloned from ...200312582135
area directly mediates nitrogen regulation of gibberellin biosynthesis in gibberella fujikuroi, but its activity is not affected by nmr.area (nit2) is a general transcription factor involved in derepression of numerous genes responsible for nitrogen utilization in gibberella fujikuroi and many other fungi. we have previously shown that the deletion of area-gf resulted in mutants with significantly reduced gibberellin (ga) production. here we demonstrate that the expression level of six of the seven ga biosynthesis genes is drastically reduced in mutants lacking area. furthermore, we show that, despite the fact that gas are nitro ...200312581353
tim50, a novel component of the tim23 preprotein translocase of mitochondria.the preprotein translocase of the inner membrane of mitochondria (tim23 complex) is the main entry gate for proteins of the matrix and the inner membrane. we isolated the tim23 complex of neurospora crassa. besides tim23 and tim17, it contained a novel component, referred to as tim50. tim50 spans the inner membrane with a single transmembrane segment and exposes a large hydrophilic domain in the intermembrane space. tim50 is essential for viability of yeast. mitochondria from cells depleted of t ...200312574118
identification of a high-affinity phosphate transporter gene in a prasinophyte alga, tetraselmis chui, and its expression under nutrient limitation.a high-affinity phosphate transporter gene, tcpho, was isolated from a growth-dependent subtracted cdna library of the marine unicellular alga tetraselmis chui. the full-length cdna of tcpho obtained by 5' and 3' rapid amplification of cdna ends was 1,993 bp long and encoded an open reading frame consisting of 610 amino acids. the deduced amino acid sequence of tcpho exhibited 51.6 and 49.8% similarity to the amino acid sequences of pho89 from saccharomyces cerevisiae and pho4 from neurospora cr ...200312570992
the internal alternative nadh dehydrogenase of neurospora crassa mitochondria.an open reading frame homologous with genes of non-proton-pumping nadh dehydrogenases was identified in the genome of neurospora crassa. the 57 kda nadh:ubiquinone oxidoreductase acts as internal (alternative) respiratory nadh dehydrogenase (ndi1) in the fungal mitochondria. the precursor polypeptide includes a pre-sequence of 31 amino acids, and the mature enzyme comprises one fad molecule as a prosthetic group. it catalyses specifically the oxidation of nadh. western blot analysis of fungal mi ...200312556227
the oligomeric state, complex formation, and chaperoning activity of hsp70 and hsp80 of neurospora crassa.molecular chaperones perform vital cellular functions under normal growth conditions and protect cells against stress-induced damage. the stress proteins hsp70 and hsp80 of neurospora crassa were extracted from heat-shocked mycelium, purified to near homogeneity, and examined with respect to their oligomeric state, complex formation, and chaperoning properties. their oligomeric state was assessed by dynamic light-scattering measurements, and both hsp70 and hsp80 were observed to form a range of ...200212555813
isolation and genetic characterization of the neurospora crassa rev1 and rev7 homologs: evidence for involvement in damage-induced mutagenesis.in a previous paper, we reported that the neurospora crassa upr-1 gene is a homolog of the yeast gene rev3, which encodes the catalytic subunit of dna polymerase zeta (polzeta). characterization of the upr-1 mutant indicated that the upr1 protein plays a role in dna repair and mutagenesis. to help understand the mechanisms of mutagenic dna repair in the n. crassa more extensively, we identified n. crassa homologs of yeast rev1 and rev7 and obtained mutants ncrev1 or ncrev7, which had similar phe ...200312547396
detection of unpaired dna at meiosis results in rna-mediated silencing.during meiosis, homologous chromosomes must pair in order to permit recombination and correct chromosome segregation to occur. two recent papers show that meiotic pairing is also important for correct gene expression during meiosis. they describe data for the filamentous fungus neurospora crassa that show that a lack of pairing generated by ectopic integration of genes can result in silencing of genes expressed during meiosis. this can result in aberrant meioses whose defects are specific to the ...200312539234
interaction of the mitotic inhibitor monastrol with human kinesin eg5.the microtubule-dependent kinesin-like protein eg5 from homo sapiens is involved in the assembly of the mitotic spindle. it shows a three-domain structure with an n-terminal motor domain, a central coiled coil, and a c-terminal tail domain. in vivo hseg5 is reversibly inhibited by monastrol, a small cell-permeable molecule that causes cells to be arrested in mitosis. both monomeric and dimeric eg5 constructs have been examined in order to define the minimal monastrol binding domain on hseg5. nmr ...200312525161
expression of protein phosphatase 1 during the asexual development of neurospora crassa.we cloned and sequenced the cdna and the gene encoding the catalytic subunit of protein phosphatase 1 from the filamentous fungus neurospora crassa. the gene, designated ppp-1 (phosphoprotein phosphatase 1), was mapped by restriction fragment length polymorphism to linkage group iii, in the vicinity of con-7 and trp-1. the expression of the gene was monitored by reverse transcriptase and polymerase chain reactions, by western blotting, and by protein phosphatase activity assays in synchronized c ...200312524044
mcp1 encodes the molybdenum cofactor carrier protein in chlamydomonas reinhardtii and participates in protection, binding, and storage functions of the cofactor.the molybdenum cofactor (moco) is essential for the activity of all molybdoenzymes except nitrogenase. the cdna for the moco carrier protein (mococp) of chlamydomonas reinhardtii has been cloned by reverse transcription pcr approaches with primers designed from microsequenced peptides of this protein. the c. reinhardtii mococp has been expressed in escherichia coli. the recombinant protein has been purified to electrophoretic homogeneity and is found assembled into a homotetramer when moco is no ...200312519777
cellular rna-dependent rna polymerase involved in posttranscriptional gene silencing has two distinct activity modes.recent genetic data suggest that proteins homologous to a plant rna-dependent rna polymerase (rdrp) play a central role in posttranscriptional gene silencing (ptgs) in many organisms. we show here that purified recombinant protein qde-1, a genetic component of ptgs ("quelling") in the fungus neurospora crassa, possesses rna polymerase activity in vitro. the full-length enzyme and its enzymatically active c-terminal fragment perform two different reactions on single-stranded rna templates, synthe ...200212504016
characterization of dna binding and the cysteine rich region of sre, a gata factor in neurospora crassa involved in siderophore synthesis.several homologous genes encoding proteins involved in regulating siderophore synthesis in fungi have been isolated, including the sre gene from the filamentous fungus neurospora crassa. we present data that further characterize sre and provide new insights into the regulation of iron homeostasis in neurospora. sre is a member of the gata factor family, which is comprised of transcription factors that contain either one or two zinc finger motifs that recognize and bind to gata-containing dna seq ...200212484767
identification and functional expression of taha, a filamentous fungal gene involved in copper trafficking to the secretory pathway in trametes versicolor.in this study, cdna and genomic clones encoding a homologue of the yeast gene anti-oxidant 1 (atx1) from the white-rot fungus trametes versicolor, a basidiomycete known to produce several laccase isoenzymes involved in lignin degradation, were identified. this gene, named trametes atx homologue (taha), encodes a protein of 7.9 kda with 56% identity to the yeast atx1p sequence. two different alleles of taha were obtained that differed mainly in their intervening sequences and in a 425 nt insertio ...200212480908
detection of mutagenic activities of various pesticides in neurospora crassa. 200312478420
a blumeria graminisf.sp. hordei bac library--contig building and microsynteny studies.a bacterial artificial chromosome (bac) library of blumeria graminis f.sp. hordei, containing 12,000 clones with an average insert size of 41 kb, was constructed. the library represents about three genome equivalents and bac-end sequencing showed a high content of repetitive sequences, making contig-building difficult. to identify overlapping clones, several strategies were used: colony hybridisation, pcr screening, fingerprinting techniques and the use of single-copy expressed sequence tags. th ...200212478389
an ensemble method for identifying regulatory circuits with special reference to the qa gene cluster of neurospora crassa.a chemical reaction network for the regulation of the quinic acid (qa) gene cluster of neurospora crassa is proposed. an efficient monte carlo method for walking through the parameter space of possible chemical reaction networks is developed to identify an ensemble of deterministic kinetics models with rate constants consistent with rna and protein profiling data. this method was successful in identifying a model ensemble fitting available rna profiling data on the qa gene cluster.200212477937
multiple functions of mfa-1, a putative pheromone precursor gene of neurospora crassa.a putative pheromone precursor gene of neurospora crassa, mfa-1 (which encodes mating factor a-1), was identified as the most abundant clone in starved mycelial and perithecial cdna libraries. northern analysis demonstrated high mfa-1 expression in all mating type a tissues and suggested low expression levels in mat a tissues. the mfa-1 gene was expressed as an approximately 1.2-kb transcript predicted to encode a 24-residue peptide, followed by a long 3' untranslated region (3' utr). the predic ...200212477799
cloning and characterization of scon-3+, a new member of the neurospora crassa sulfur regulatory system.the sulfur regulatory system of neurospora crassa consists of a group of sulfur-regulated structural genes (e.g., arylsulfatase) that are under coordinate control of the cys3 positive regulator and sulfur controller (scon) negative regulators. here we report on the cloning of scon-3(+), which encodes a polypeptide of 171 amino acids and is a skp1 family homolog. repeat-induced point mutation of scon-3(+) resulted in a phenotype of constitutive expression of arylsulfatase, a phenotype consistent ...200212477788
shared and independent roles for a galpha(i) protein and adenylyl cyclase in regulating development and stress responses in neurospora crassa.growth and development are regulated using cyclic amp (camp)-dependent and -independent pathways in neurospora crassa. the cr-1 adenylyl cyclase mutant lacks detectable camp and exhibits numerous defects, including colonial growth habit, short aerial hyphae, premature conidiation on plates, inappropriate conidiation in submerged culture, and increased thermotolerance. evidence suggests that the heterotrimeric galpha protein gna-1 is a direct positive regulator of adenylyl cyclase. deltagna-1 str ...200212456011
colletotrichum trifolii tb3 kinase, a cot1 homolog, is light inducible and becomes localized in the nucleus during hyphal elongation.colletotrichum trifolii is a fungal pathogen responsible for anthracnose disease of alfalfa. previously, a serine/threonine protein kinase gene from this fungus (tb3), which is a functional homolog of the neurospora crassa cot1 kinase, has been isolated in our laboratory and appears to be associated with hyphal elongation and branching. in this report we show that light treatment rapidly induces tb3 expression and hyphal branching frequency. western analysis showed tb3 localization in both the c ...200212456010
characterization of inhibitor-resistant histone deacetylase activity in plant-pathogenic fungi.hc-toxin, a cyclic peptide made by the filamentous fungus cochliobolus carbonum, is an inhibitor of histone deacetylase (hdac) from many organisms. it was shown earlier that the hdac activity in crude extracts of c. carbonum is relatively insensitive to hc-toxin as well as to the chemically unrelated hdac inhibitors trichostatin and d85, whereas the hdac activity of aspergillus nidulans is sensitive (g. brosch et al., biochemistry 40:12855-12863, 2001). here we report that hc-toxin-resistant hda ...200212456002
a g-protein beta subunit required for sexual and vegetative development and maintenance of normal g alpha protein levels in neurospora crassa.the genome of the filamentous fungus neurospora crassa contains a single gene encoding a heterotrimeric g-protein beta subunit, gnb-1. the predicted gnb-1 protein sequence is most identical to g beta proteins from the filamentous fungi cryphonectria parasitica and aspergillus nidulans. n. crassa gnb-1 is also 65% identical to the human gnb-1 protein but only 38 and 45% identical to g beta proteins from budding and fission yeasts. previous studies in animal and fungal systems have elucidated phen ...200212455986
neurospora clock-controlled gene 9 (ccg-9) encodes trehalose synthase: circadian regulation of stress responses and development.the circadian clock of neurospora crassa regulates the rhythmic expression of a number of genes encoding diverse functions which, as an ensemble, are adaptive to life in a rhythmic environment of alternating levels of light and dark, warmth and coolness, and dryness and humidity. previous differential screens have identified a number of such genes based solely on their cycling expression, including clock-controlled gene 9 (ccg-9). sequence analysis now shows the predicted ccg-9 polypeptide to be ...200212455969
distinct signaling pathways from the circadian clock participate in regulation of rhythmic conidiospore development in neurospora crassa.several different environmental signals can induce asexual spore development (conidiation) and expression of developmentally regulated genes in neurospora crassa. however, under constant conditions, where no environmental cues for conidiation are present, the endogenous circadian clock in n. crassa promotes daily rhythms in expression of known developmental genes and of conidiation. we anticipated that the same pathway of gene regulation would be followed during clock-controlled conidiation and ...200212455961
aspergillus nidulans catalase-peroxidase gene (cpea) is transcriptionally induced during sexual development through the transcription factor stua.catalases, peroxidases, and catalase-peroxidases are important enzymes to cope with reactive oxygen species in pro- and eukaryotic cells. in the filamentous fungus aspergillus nidulans three monofunctional catalases have been described, and a fourth catalase activity was observed in native polyacrylamide gels. the latter activity is probably due to the bifunctional enzyme catalase-peroxidase, which we characterized here. the gene, named cpea, encodes an 81-kda polypeptide with a conserved motif ...200212455692
effects of iprodione and fludioxonil on glycerol synthesis and hyphal development in candida albicans.we investigated the effects of iprodione and fludioxonil on the pathogenic yeast candida albicans. growth of the wild-type ifo1385 strain of c. albicans was inhibited by both fungicides, while saccharomyces cerevisiae was basically unaffected by them even at a concentration of 25 microg/ml. both fungicides stimulated glycerol synthesis in c. albicans but not in s. cerevisiae. the antioxidant alpha-tocopherol acetate and the cytochrome p-450 inhibitor piperonyl butoxide antagonized the fungitoxic ...200212450134
morphological alterations induced by cold-shock in neurospora crassa.the effects of cold-shock on neurospora crassa was here investigated. in cold-shock treated hyphae (6 h at 4 degrees c) the branching pattern was changed and dichotomy as well as a fan-like-pattern structure were temporarily detected at the tips. when exogenous ca(2+) (500 mm) was added to the hyphae, these morphological alterations were not detected. in addition, the protein electrophoretic profile of cultures submitted to cold-shock was different from that of cultures kept at 30 degrees c. aft ...200212442300
a non-modular type b feruloyl esterase from neurospora crassa exhibits concentration-dependent substrate inhibition.feruloyl esterases, a subclass of the carboxylic acid esterases (ec 3.1.1.1), are able to hydrolyse the ester bond between the hydroxycinnamic acids and sugars present in the plant cell wall. the enzymes have been classified as type a or type b, based on their substrate specificity for aromatic moieties. we show that neurospora crassa has the ability to produce multiple ferulic acid esterase activities depending upon the length of fermentation with either sugar beet pulp or wheat bran substrates ...200312435269
an ip3-activated ca2+ channel regulates fungal tip growth.hyphal extension in fungi requires a tip-high ca(2+) gradient, which is generated and maintained internally by inositol (1,4,5)-trisphosphate (ip(3))-induced ca(2+) release from tip-localized vesicles and subapical ca(2+) sequestration. using the planar bilayer method we demonstrated the presence of two types of ip(3)-activated ca(2+) channels in neurospora crassa membranes with different conductances: one low (13 picosiemens), the other high (77 picosiemens). on sucrose density gradients the lo ...200212432087
analysis of three separate probes suggests the absence of endocytosis in neurospora crassa hyphae.reports of the existence of endocytosis in filamentous fungi have been conflicting and inconclusive. for this reason, we have tested three independent markers in neurospora crassa: the electron opaque marker lanthanum (la) and the fluorescent probes lucifer yellow (ly) and fm4-64. both la and ly were endocytosed by saccharomyces cerevisiae cells, which were used as positive controls for endocytosis, but the probes did not accumulate in n. crassa hyphae. only fm4-64 became internalized into n. cr ...200212431457
evidence for moea-dependent formation of the molybdenum cofactor from molybdate and molybdopterin in escherichia coli.the function of the moea protein in the biosynthesis of the molybdenum cofactor (moco) was analyzed in vitro, using purified his(6)-moea from escherichia coli, molybdopterin (mpt) isolated from buttermilk xanthine oxidase and molybdate. the formation of moco was monitored by the reconstitution of nitrate reductase activity in extracts of the neurospora crassa nit-1 mutant. formation of moco from mpt and molybdate required moea and l-cysteine or glutathione. the reaction proceeded at micromolar m ...200212420167
validation of cdc68p as a novel antifungal target.candida albicans is the main cause of systemic fungal infections for which there is an urgent need for novel antifungal drugs. the cp (cdc68p-pob3p) complex, which is involved in transcription elongation, was evaluated as a putative antifungal target. in order to predict the consequences of inhibition of this complex, the largest cp subunit in saccharomyces cerevisiae, cdc68p, was the first novel target to be tested in gate, a recently described, quantitative target inactivation system. depletio ...200212420162
the effects of ropy-1 mutation on cytoplasmic organization and intracellular motility in mature hyphae of neurospora crassa.we have used light and electron microscopy to document the cytoplasmic effects of the ropy (ro-1) mutation in mature hyphae of neurospora crassa and to better understand the role(s) of dynein during hyphal tip growth. based on video-enhanced dic light microscopy, the mature, growing hyphae of n. crassa wild type could be divided into four regions according to cytoplasmic organization and behavior: the apical region (i) and three subapical regions (ii, iii, and iv). a well-defined spitzenkörper d ...200212409101
a fourier transform infrared study of neurospora rhodopsin: similarities with archaeal rhodopsins.the nop-1 gene from the eukaryote neurospora crassa, a filamentous fungus, has recently been shown to encode an archaeal rhodopsin-like protein nop-1. to explore the functional mechanism of nop-1 and its possible similarities to archaeal and visual rhodopsins, static and time-resolved fourier transform infrared difference spectra were measured from wild-type nop-1 and from a mutant containing an asp-->glu substitution in the schiff base (sb) counterion, asp131 (d131e). several conclusions could ...200212403457
characterization of neurospora crassa tom40-deficient mutants and effect of specific mutations on tom40 assembly.the tom complex (translocase of the outer mitochondrial membrane) is responsible for the recognition of mitochondrial preproteins synthesized in the cytosol and for their translocation across or into the outer mitochondrial membrane. tom40 is the major component of the tom complex and forms the translocation pore. we have created a tom40 mutant of neurospora crassa and have demonstrated that the gene is essential for the viability of the organism. mitochondria with reduced levels of tom40 were d ...200312399467
the trichoderma atroviride seb1 (stress response element binding) gene encodes an agggg-binding protein which is involved in the response to high osmolarity stress.the chitinase genes of trichoderma spp. (ech42, chit33, nag1) contain one or more copies of a pentanucleotide element (5'-agggg-3') in their 5'-noncoding regions. in saccharomyces cerevisiae, this motif is recognized and bound by the stress response regulator proteins msn2p/msn4p. to test whether this motif in the chitinase promoters is bound by a trichoderma msn2/4p homolog, we have cloned a gene (seb1) from t. atroviride which encodes a c2h2 zinc-finger protein that is 62 (64)% identical to s. ...200212395196
modeling the three-dimensional structure of h+-atpase of neurospora crassa.homology modeling in combination with transmembrane topology predictions are used to build the atomic model of neurospora crassa plasma membrane h+-atpase, using as template the 2.6 a crystal structure of rabbit sarcoplasmic reticulum ca2+-atpase [toyoshima, c., nakasako, m., nomura, h. & ogawa, h. (2000) nature 405, 647-655]. comparison of the two calcium-binding sites in the crystal structure of ca2+-atpase with the equivalent region in the h+-atpase model shows that the latter is devoid of mo ...200212392557
definition of the nuclear encoded protein composition of bovine heart mitochondrial complex i. identification of two new subunits.mitochondrial nadh:ubiquinone oxidoreductase (complex i) from bovine heart is a complicated multisubunit, membrane-bound assembly. seven subunits are encoded by mitochondrial dna, and the sequences of 36 nuclear encoded subunits have been described. the subunits of complex i and two subcomplexes (ialpha and ibeta) were resolved on one- and two-dimensional gels and by reverse-phase high performance liquid chromatography. mass spectrometric analysis revealed two previously unknown subunits in comp ...200212381726
the transition between active and de-activated forms of nadh:ubiquinone oxidoreductase (complex i) in the mitochondrial membrane of neurospora crassa.the mammalian mitochondrial nadh:ubiquinone oxidoreductase (complex i) has been shown to exist in two kinetically and structurally distinct slowly interconvertible forms, active (a) and de-activated (d) [vinogradov and grivennikova (2001) iubmb life 52, 129-134]. this work was undertaken to investigate the putative complex i a-d transition in the mitochondrial membrane of the lower eukaryote neurospora crassa and in plasma membrane of the prokaryote paracoccus denitrificans, organisms that are e ...200312379145
a new class of tetraspanins in fungi.tetraspanins are animal proteins involved in membrane complexes that are involved in cell adhesion, differentiation, and motility. the pls1 gene from rice blast fungus magnaporthe grisea encodes a protein (pls1p) structurally related to tetraspanins that is required for pathogenicity. in botrytis cinerea public sequences, we identified an est homologous to pls1. using degenerated oligonucleotides, we amplified sequences homologous to pls1 in fungi colletotrichum lindemuthianum and neurospora cra ...200212372414
acyl derivatives of homoserine as substrates for homocysteine synthesis in neurospora crassa, yeast, and escherichia coli. 196712325384
a large ca2+-dependent channel formed by recombinant adp/atp carrier from neurospora crassa resembles the mitochondrial permeability transition pore.strong support for the central role of the adp/atp carrier (aac) in the mitochondrial permeability transition (mpt) is provided by the single-channel current measurements in patch-clamp experiments with the isolated reconstituted aac. in previous work [brustovetsky, n., and klingenberg, m. (1996) biochemistry 35, 8483-8488], this technique was applied to the aac isolated from bovine heart mitochondria. here we used recombinant aac (raac) from neurospora crassa expressed in e. coli, since aac fro ...200212269823
mutants of chlamydomonas with aberrant responses to sulfur deprivation.in the absence of sulfur, chlamydomonas reinhardtii, a unicellular green alga, increases its rate of sulfate import and synthesizes several periplasmic proteins, including an arylsulfatase (ars). these changes appear to help cells acclimate to a sulfur-deficient environment. the elevated rate of sulfate import results from an increase in the capacity and affinity of the transport system for sulfate. the synthesis of ars, a periplasmic enzyme that cleaves sulfate from aromatic compounds, enables ...199412244220
a new wc-1 mutant of neurospora crassa shows unique light sensitivity in the circadian conidiation rhythm.a new clock mutant ( rhy-2) was isolated by dna insertion mutagenesis using a plasmid that contains a region located upstream of the cmd gene in the genome of neurospora crassa. this mutant is arrhythmic with regard to conidiation in continuous darkness but rhythmic under a light-dark cycle. after plasmid rescue from genomic dna of the rhy-2 strain, the insertion was localized to the gene white collar-1 ( wc-1). plasmid dna was inserted 3' to the sequence encoding the polyglutamine region of the ...200212242499
identification of vib-1, a locus involved in vegetative incompatibility mediated by het-c in neurospora crassa.a non-self-recognition system called vegetative incompatibility is ubiquitous in filamentous fungi and is genetically regulated by het loci. different fungal individuals are unable to form viable heterokaryons if they differ in allelic specificity at a het locus. to identify components of vegetative incompatibility mediated by allelic differences at the het-c locus of neurospora crassa, we isolated mutants that suppressed phenotypic aspects of het-c vegetative incompatibility. three deletion mut ...200212242225
the hypocrea jecorina gal10 (uridine 5'-diphosphate-glucose 4-epimerase-encoding) gene differs from yeast homologues in structure, genomic organization and expression.as part of a comprehensive study on lactose metabolism in hypocrea jecorina (anamorph: trichoderma reesei), a genomic clone of the gal10 gene encoding h. jecorina uridine 5'-diphosphate (udp)-glucose 4-epimerase has been cloned and sequenced. it contains an open reading frame of 1548-base pair, interrupted by three introns, and encoding a 370-amino acids protein with similarity to pro- and eukaryotic udp-glucose-4-epimerases. h. jecorina gal10 does not contain the c-terminal mutarotase domain wh ...200212242021
nonself recognition is mediated by het-c heterocomplex formation during vegetative incompatibility.nonself recognition during vegetative growth in filamentous fungi is mediated by heterokaryon incompatibility (het) loci. in neurospora crassa, het-c is one of 11 het loci. three allelic specificity groups, termed het-c(or), het-c(pa) and het-c(gr), exist in natural populations. heterokaryons or partial diploids that contain het-c alleles of alternative specificity show severe growth inhibition, repression of conidiation and hyphal compartmentation and death (hcd). using epitope-tagged het-c, we ...200212234924
only the mature form of the plastidic chorismate synthase is enzymatically active.coding regions of a cdna for precursor and mature chorismate synthase (cs), a plastidic enzyme, from corydalis sempervirens were expressed in escherichia coli as translational fusions to glutathione-s-transferase. fusion proteins were purified, and precursor and mature forms of cs were then released by proteolytic cleavage with factor xa. although mature cs was enzymatically active after release, activity could be detected neither for the precursor cs nor for corresponding glutathione-s-transfer ...199512228531
live-cell imaging of vegetative hyphal fusion in neurospora crassa.the process of hyphal fusion (anastomosis) in growing colonies of neurospora crassa, stained with the membrane-selective dyes fm1-43 and fm4-64, was visualized by confocal microscopy. time-lapse, live-cell imaging illustrated the dynamics of hyphal growth and anastomosis during its pre-contact, contact and post-contact, and post-fusion stages. fusion-competent hyphae were morphologically distinct and exhibited remote sensing, resulting in branch initiation and/or re-direction of growth to facili ...200212223195
the vea gene activates sexual development in aspergillus nidulans.the previously isolated gene complementing the vea1 mutation was confirmed to be the vea gene. the determined nucleotide sequence of the gene demonstrated that there is an open reading frame (orf) of a 573 amino acid polypeptide. the nucleotide sequence matched some clones of which functions were not assigned yet and the amino acid sequence matched that of neurospora crassa vea-related protein with 61% similarity. the nucleotide sequence of the vea1 mutant gene differed from that of the wild typ ...200212223191
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