structure and function analysis of the calcium-related gene spray in neurospora crassa.the spray gene was cloned, and wildtype and mutant alleles were sequenced. spray(+) has a 3452-bp open reading frame plus seven introns. the spray mutant had a t --> g transversion close to the carboxyl end, creating a stop codon (tga). the sequence shows no match to genes of known function, but the carboxyl end shows seven transmembrane domains and matches putative membrane proteins of yeast. the most abundant transcript detected was 4.4 kb in size. repeat-induced point mutagenesis produced the ...200111343401
identification of two tetranuclear fes clusters on the ferredoxin-type subunit of nadh:ubiquinone oxidoreductase (complex i).the proton-translocating nadh:ubiquinone oxidoreductase of respiratory chains (complex i) contains one flavin mononucleotide and five epr-detectable iron-sulfur clusters as redox groups. because of the number of conserved motifs typical for binding iron-sulfur clusters and the high content of iron and acid-labile sulfide of complex i preparations, it is predicted that complex i contains additional clusters which have not yet been detected by epr spectroscopy. to search for such clusters, we used ...200111352750
genome quality control: rip (repeat-induced point mutation) comes to (repeat-induced point mutation) is a silencing process discovered in neurospora crassa and so far clearly established only in this species as a currently occurring process. rip acts premeiotically on duplicated sequences, resulting in c-g to t-a mutations, with a striking preference for cpa/tpg dinucleotides. in podospora anserina, an rip-like event was observed after several rounds of sexual reproduction in a strain with a 40 kb tandem duplication resulting from homologous integration of a ...200111359565
structural requirements of tom40 for assembly into preexisting tom complexes of mitochondria.tom40 is the major subunit of the translocase of the outer mitochondrial membrane (the tom complex). to study the assembly pathway of tom40, we have followed the integration of the protein into the tom complex in vitro and in vivo using wild-type and altered versions of the neurospora crassa tom40 protein. upon import into isolated mitochondria, tom40 precursor proteins lacking the first 20 or the first 40 amino acid residues were assembled as the wild-type protein. in contrast, a tom40 precurso ...200111359915
protein-dependent transition states for ribonucleoprotein assembly.native folding and splicing by the saccharomyces cerevisiae mitochondrial bi5 group i intron rna is facilitated by both the s. cerevisiae cbp2 and neurospora crassa cyt-18 protein cofactors. both protein-bi5 rna complexes splice at similar rates, suggesting that the rna active site structure is similar in both ribonucleoproteins. in contrast, the two proteins assemble with the bi5 rna by distinct mechanisms and bind opposing, but partially overlapping, sides of the group i intron catalytic core. ...200111399081
tom40, the pore-forming component of the protein-conducting tom channel in the outer membrane of mitochondria.tom40 is the main component of the preprotein translocase of the outer membrane of mitochondria (tom complex). we have isolated tom40 of neurospora crassa by removing the receptor tom22 and the small tom components tom6 and tom7 from the purified tom core complex. tom40 is organized in a high molecular mass complex of approximately 350 kd. it forms a high conductance channel. mitochondrial presequence peptides interact specifically with tom40 reconstituted into planar lipid membranes and decreas ...200111402060
photochemical reaction cycle and proton transfers in neurospora was recently found that nop-1, a membrane protein of neurospora crassa, shows homology to haloarchaeal rhodopsins and binds retinal after heterologous expression in pichia pastoris. we report on spectroscopic properties of the neurospora rhodopsin (nr). the photocycle was studied with flash photolysis and time-resolved fourier-transform infrared spectroscopy in the ph range 5-8. proton release and uptake during the photocycle were monitored with the ph-sensitive dye, pyranine. kinetic and spe ...200111435422
molecular cloning of the rps0 gene from candida tropicalis.the saccharomyces cerevisiae rps0 a and b genes encode proteins essential for maturation of the 40s ribosomal subunit precursors. we have isolated a homologue of the rps0 gene from candida tropicalis, which we named ctrps0. the c. tropicalis rps0 encodes a protein of 261 amino acid residues with a predicted molecular weight of 28.65 kda and an isoelectric point of 4.79. ctrps0p displays significant amino acid sequence homology with rps0p from c. albicans, s. cerevisiae, neurospora crassa, schizo ...200111447603
identification of the gene at the pmg locus, encoding system ii, the general amino acid transporter in neurospora crassa.mutations at the pmg locus in neurospora crassa cause a deficiency in a transport system for a broad range of amino acids. we have isolated a gene that encodes a protein with a high degree of sequence similarity to the gap1 general amino acid permease in saccharomyces cerevisiae. our data indicate that this is the gene at the pmg locus. it encodes a 572-residue protein with a molecular mass of 62,649 da. the predicted secondary structure has 12 membrane-spanning regions, a feature characteristic ...200111456465
regions of microsynteny in magnaporthe grisea and neurospora crassa.a bacterial artificial chromosome (bac) clone containing 110,467 bp of genomic dna from magnaporthe grisea was sequenced, annotated, and compared to the genomes of neurospora crassa, candida albicans, and saccharomyces cerevisiae. twenty-six open reading frames (orfs), involved in multiple biochemical pathways, were identified in the bac sequence. a region of 53 kb, containing 18 of the 26 orfs, was found to be syntenic to a portion of the n. crassa genome. subregions of complete colinearity as ...200111456466
dna methylation inhibits expression and transposition of the neurospora tad retrotransposon.tad is a line-like retrotransposon of the filamentous fungus neurospora crassa. we have analyzed both expression and transposition of this element using strains with a single copy of tad located in the 5' noncoding sequences of the am (glutamate dehydrogenase) gene. tad in this position has been shown to carry a de novo cytosine methylation signal which causes reversible methylation of both tad and am upstream sequences. here we find that methylation of the tad sequences inhibits both tad expres ...200111459196
the methionine biosynthetic pathway from homoserine in pseudomonas putida involves the metw, metx, metz, meth and mete gene products.biosynthesis of methionine from homoserine in pseudomonas putida takes place in three steps. the first step is the acylation of homoserine to yield an acyl-l-homoserine. this reaction is catalyzed by the products of the metxw genes and is equivalent to the first step in enterobacteria, gram-positive bacteria and fungi, except that in these microorganisms the reaction is catalyzed by a single polypeptide (the product of the meta gene in escherichia coli and the met5 gene product in neurospora cra ...200111479715
evidence that fungal mep proteins mediate diffusion of the uncharged species nh(3) across the cytoplasmic membrane.methylammonium and ammonium (mep) permeases of saccharomyces cerevisiae belong to a ubiquitous family of cytoplasmic membrane proteins that transport only ammonium (nh(4)(+) + nh(3)). transport and accumulation of the ammonium analog [(14)c]methylammonium, a weak base, led to the proposal that members of this family were capable of energy-dependent concentration of the ammonium ion, nh(4)(+). in bacteria, however, atp-dependent conversion of methylammonium to gamma-n-methylglutamine by glutamine ...200111486013
ca(2+) shuttling in vesicles during tip growth in neurospora crassa.tip-growing organisms maintain an apparently essential tip-high gradient of cytoplasmic ca(2+). in the oomycete saprolegnia ferax, in pollen tubes and root hairs, the gradient is produced by a tip-localized ca(2+) influx from the external medium. such a gradient is normally dispensable for neurospora crassa hyphae, which may maintain their ca(2+) gradient by some form of internal recycling. we localized ca(2+) in n. crassa hyphae at the ultrastructural level using two techniques (a) electron spe ...200111495575
functional analysis of pathogenicity genes in a genomics world.genome-wide mutational and expression analyses have been performed in yeast and provide a model for large-scale analysis of gene function in filamentous fungi. the recent completion of the neurospora crassa genome offers a resource for comparative analysis with plant pathogenic filamentous fungi. these advances have important implications for molecular genetic studies of pathogenicity genes.200111495799
a family of dna repeats in aspergillus nidulans has assimilated degenerated the course of a chromosomal walk towards the centromere of chromosome iv of aspergillus nidulans, several cross-hybridizing genomic cosmid clones were isolated. restriction mapping of two such clones revealed that their restriction patterns were similar in a region of at least 15 kb, indicating the presence of a large repeat. the nature of the repeat was further investigated by sequencing and southern analysis. the study revealed a family of long dispersed repeats with a high degree of sequen ...200111523805
the grp78 promoter of neurospora crassa: constitutive, stress and differentiation-dependent protein-binding order to characterize the transcriptional regulation of the grp78 (glucose-regulated protein 78 kda) gene of neurospora crassa, a 1.65-kb genomic fragment upstream of the protein-coding region was sequenced and analysed. a single transcription start point was mapped 160 nt upstream of the first codon. several distinct protein-dna binding sites were identified in the promoter region by non-radioactive scanning electrophoretic mobility shift analysis during growth, stress treatment and differen ...200111525405
spectroscopic and kinetic characterization of the bifunctional chorismate synthase from neurospora crassa: evidence for a common binding site for 5-enolpyruvylshikimate 3-phosphate and nadph.chorismate synthase catalyzes the anti-1,4-elimination of the phosphate group and the c-(6pror) hydrogen from 5-enolpyruvylshikimate 3-phosphate to yield chorismate, a central building block in aromatic amino acid biosynthesis. the enzyme has an absolute requirement for reduced fmn, which in the case of the fungal chorismate synthases is supplied by an intrinsic fmn:nadph oxidoreductase activity, i.e. these enzymes have an additional catalytic activity. therefore, these fungal enzymes have been ...200111526120
a new yeast metabolon involving at least the two first enzymes of arginine biosynthesis: acetylglutamate synthase activity requires complex formation with acetylglutamate reading frame yjl071w of saccharomyces cerevisiae was shown to be arg2 and identified as the structural gene for acetylglutamate synthase, first step in arginine biosynthesis. the three ascomycete acetylglutamate synthases characterized to date appear homologous, but unlike the other enzymes of the yeast arginine biosynthesis pathway, they showed no significant similarity to their prokaryotic equivalents. the measured synthase activity did not increase with the number of arg2 gene copies un ...200111553611
map-1 and iap-1, two novel aaa proteases with catalytic sites on opposite membrane surfaces in mitochondrial inner membrane of neurospora crassa.eukaryotic aaa proteases form a conserved family of membrane-embedded atp-dependent proteases but have been analyzed functionally only in the yeast saccharomyces cerevisiae. here, we have identified two novel members of this protein family in the filamentous fungus neurospora crassa, which were termed map-1 and iap-1. both proteins are localized to the inner membrane of mitochondria. they are part of two similar-sized high molecular mass complexes, but expose their catalytic sites to opposite me ...200111553723
a fungal perspective on human inborn errors of metabolism: alkaptonuria and beyond.crucial for the establishment and development of biochemical genetics as a self-standing discipline was beadle and tatum's choice of neurospora crassa as experimental organism some 60 years ago. although garrod's insights on biochemical genetics and his astonishingly modern concepts of biochemical individuality and susceptibility to disease had been ignored by their contemporaries, beadle acknowledged on several occasions how close garrod had come to the "one-gene-one-enzyme" hypothesis. in an u ...200111567547
transcriptional regulation of the nitrate reductase gene in chlorella vulgaris: identification of regulatory elements controlling expression.nitrate reductase (nr), the rate-limiting and primary control point of the nitrate assimilation pathway, is regulated at transcriptional and post-transcriptional levels. to better understand how nr is regulated at the transcriptional level in chlorella vulgaris, studies were performed to identify the factors regulating nr expression. sequence analysis of the nr promoter identified a number of potential sites that were investigated by mobility shift assays. of the protein-binding sites found, sev ...200111680822
cloning and sequence analysis of cnaa gene encoding the catalytic subunit of calcineurin from aspergillus oryzae.calcineurin has been implicated in ion-homeostasis, stress adaptation in yeast and for hyphal growth in filamentous fungi. genomic dna and cdna encoding the catalytic subunit of calcineurin (cnaa) were isolated from aspergillus oryzae. the cnaa open reading frame extended to 1727 bp and encoded a putative protein of 514 amino acids. comparative analysis of the nucleotide sequence of cnaa genomic dna and cdna confirmed the presence of three introns and a highly conserved calmodulin binding domain ...200111682197
unusual properties of the fungal conventional kinesin neck domain from neurospora crassa.fungal conventional kinesins are unusually fast microtubule motor proteins. to compare the functional organization of fungal and animal conventional kinesins, a set of c-terminal deletion mutants of the neurospora crassa conventional kinesin, nckin, was investigated for its biochemical and biophysical properties. while the shortest, monomeric construct comprising the catalytic core and the neck-linker (nckin343) displays very high steady-state atpase (k(cat) = 260/s), constructs including both t ...200111707394
conversion of pipecolic acid into lysine in penicillium chrysogenum requires pipecolate oxidase and saccharopine reductase: characterization of the lys7 gene encoding saccharopine reductase.pipecolic acid is a component of several secondary metabolites in plants and fungi. this compound is useful as a precursor of nonribosomal peptides with novel pharmacological activities. in penicillium chrysogenum pipecolic acid is converted into lysine and complements the lysine requirement of three different lysine auxotrophs with mutations in the lys1, lys2, or lys3 genes allowing a slow growth of these auxotrophs. we have isolated two p. chrysogenum mutants, named 7.2 and 10.25, that are una ...200111717275
partial amino acid sequences of peptidyl-prolyl isomerases offusarium sporotrichioides.peptidyl-proprylyl cis-trans isomerase (ppiase) activity was observed from crude extract of fusarium sporotrichioides. proteins from this fungi were separated by two-dimensional polyacrylamide gel electrophoresis and more than one thousand protein spots were separated. two cytosolic ppiases were found by the n-terminal sequencing from the two separated spots. the n-terminal 41 residues of the major protein spot showed high sequence identity (63.4%) with ppiase from neurospora crassa. this protei ...199511725072
molecular and kinetic study of catalase-1, a durable large catalase of neurospora crassa.catalase-1 (cat-1), one of the two monofunctional catalases of neurospora crassa, increases during asexual spore formation to constitute 0.6% of total protein in conidia. cat-1 was purified 170-fold with a yield of 48% from conidiating cultures. like most monofunctional catalases, cat-1 is a homotetramer, resistant to inactivation by solvents, fully active over a ph range of 4-12, and inactivated by 3-amino-1,2,4-triazole. unlike most monofunctional catalases, cat-1 consists of 88 kda monomers t ...200111728803
homologous expression and characterization of cel61a (eg iv) of trichoderma reesei.there are currently four proteins in family 61 of the glycoside hydrolases, from trichoderma reesei, agaricus bisporus, cryptococcus neoformans and neurospora crassa. the enzymatic activity of these proteins has not been studied thoroughly. we report here the homologous expression and purification of t. reesei cel61a [previously named endoglucanase (eg) iv]. the enzyme was expressed in high amounts with a histidine tag on the c-terminus and purified by metal affinity chromatography. this is the ...200111737205
identification of genes in the bw/be regulatory cascade in ustilago the phytopathogenic fungus ustilago maydis, the switch to filamentous growth and pathogenic development is controlled by a heterodimeric transcription factor consisting of the bw and be homeodomain proteins. to identify genes in the regulatory cascade triggered by the bw/be heterodimer, we have constructed strains in which transcription of the b genes is inducible by either arabinose or nitrate. at different time-points after induction, genes that are switched on or off were identified throug ...200111737646
identification of transcriptionally expressed pheromone receptor genes in filamentous ascomycetes.detection of pheromone genes in filamentous ascomycetes implicated the presence of pheromone receptor genes. similar to yeasts and basidiomycetes, these might be involved in a g-protein triggered signal transduction pathway during mating. we have identified two pheromone receptor genes, named pre1 and pre2, in the genome of the heterothallic filamentous ascomycete neurospora crassa and the closely related homothallic sordaria macrospora. the deduced pre1 gene product is a putative seven-transmem ...200111738813
amplified uvra protein can ameliorate the ultraviolet sensitivity of an escherichia coli reca mutant.when a reca strain of escherichia coli was transformed with the multicopy plasmid psf11 carrying the uvra gene of e. coli, its extreme ultraviolet (uv) sensitivity was decreased. the sensitivity of the lexa1 (ind(-)) strain to uv was also decreased by psf11. the reca cells expressing neurospora crassa uv damage endonuclease (uvde), encoding uv-endonuclease, show uv resistance. on the other hand, only partial amelioration of uv sensitivity of the reca strain was observed in the presence of the pl ...200111738941
mips: a database for genomes and protein sequences.the munich information center for protein sequences (mips-gsf, neuherberg, germany) continues to provide genome-related information in a systematic way. mips supports both national and european sequencing and functional analysis projects, develops and maintains automatically generated and manually annotated genome-specific databases, develops systematic classification schemes for the functional annotation of protein sequences, and provides tools for the comprehensive analysis of protein sequence ...200211752246
the role of g protein alpha subunits in the infection process of the gray mold fungus botrytis identify signal transduction pathways of the gray mold fungus botrytis cinerea involved in host infection, we used heterologous hybridization and a polymerase chain reaction (pcr)-based approach to isolate two genes (bcg1 and bcg2) encoding alpha subunits of heterotrimeric gtp-binding proteins. both genes have homologues in other fungi: bcg1 is a member of the g alpha(i) class, whereas bcg2 has similarities to the magc gene of magnaporthe grisea and the gna-2 gene of neurospora crassa. revers ...200111763127
analysis of genes expressed during rice-magnaporthe grisea interactions.expressed sequence tag (est) analysis was applied to identify rice genes involved in defense responses against infection by the blast fungus magnaporthe grisea and fungal genes involved in growth within the host during a compatible interaction. a total of 511 clones was sequenced from a cdna library constructed from rice leaves (oryza sativa cv. nipponbare) infected with m. grisea strain 70-15 to generate 296 nonredundant ests. the sequences of 293 clones (57.3%) significantly matched national c ...200111763134
protein import into mitochondria.most mitochondrial proteins are encoded by the nuclear genome and thus have to be imported into mitochondria from the cytosol. protein translocation across and into the mitochondrial membranes is a multistep process facilitated by the coordinated action of at least four specialized translocation systems in the outer and inner membranes of mitochondria. the outer membrane contains one general translocase, the tom complex, whereas three distinct translocases are located in the inner membrane, whic ...200111798021
targeting vectors for gene diversification by meiotic recombination in neurospora crassa.we have constructed a pair of vectors, pdv2 and pdv3, that enable targeted insertion of exogenous dna into linkage group i of neurospora crassa at the his-3 locus. transplaced sequences are inserted between his-3 and the cog(l) recombination hot spot and include his-3 mutations that allow meiotic recombination initiated by cog(l) to be monitored. selection of correctly placed transforming dna is based on complementation between different his-3 alleles borne by the plasmids and transformation hos ...200211798282
osmoregulation and fungicide resistance: the neurospora crassa os-2 gene encodes a hog1 mitogen-activated protein kinase homologue.neurospora crassa osmosensitive (os) mutants are sensitive to high osmolarity and therefore are unable to grow on medium containing 4% nacl. we found that os-2 and os-5 mutants were resistant to the phenylpyrrole fungicides fludioxonil and fenpiclonil. to understand the relationship between osmoregulation and fungicide resistance, we cloned the os-2 gene by using sib selection. os-2 encodes a putative mitogen-activated protein (map) kinase homologous to hog1 and can complement the osmosensitive ...200211823187
the oxa1 protein forms a homooligomeric complex and is an essential part of the mitochondrial export translocase in neurospora crassa.the oxa1 protein is a ubiquitous constituent of the inner membrane of mitochondria. oxa1 was identified in yeast as a crucial component of the protein export machinery known as the oxa translocase, which facilitates the integration of proteins from the mitochondrial matrix into the inner membrane. we have identified the neurospora crassa oxa1 protein which shows a sequence identity of 22% to the yeast homologue. despite the low level of identity, the function of the homologues is conserved as th ...200211823466
identification and characterization of novel cytochrome p450 genes from the white-rot basidiomycete, coriolus versicolor.using a reverse-transcription polymerase chain reaction (rt-pcr) technique, cytochrome p450 genes were cloned from the lignin-degrading basidiomycete, coriolus versicolor. one possible p450 gene was identified, which consisted of 1,672 nucleotides and a poly(a) tail and encoded a deduced protein containing 449 amino acids. the deduced amino acid sequence revealed the presence of the p450 heme-binding motif, strongly suggesting that this protein belongs to the p450 superfamily, then designated cy ...200211831480
ds-like restless deletion derivatives occur in tolypocladium inflatum and two foreign hosts, neurospora crassa and penicillium chrysogenum.single copies of the transposon restless from tolypocladium inflatum were introduced into neurospora crassa and penicillium chrysogenum. excision of restless from its donor site was investigated in n. crassa and in p. chrysogenum using direct selective conditions. in n. crassa, forward selection was also analyzed. deleted restless elements were frequently obtained in addition to the expected complete removal of restless from its donor site. similar deleted elements were also identified in t. inf ...200211848679
trna-like recognition of group i introns by a tyrosyl-trna synthetase.the neurospora crassa mitochondrial tyrosyl-trna synthetase (cyt-18 protein) functions in splicing group i introns by promoting the formation of the catalytically active rna structure. previous work suggested that cyt-18 recognizes a conserved trna-like structure of the group i intron catalytic core. here, directed hydroxyl-radical cleavage assays show that the nucleotide-binding fold and c-terminal domains of cyt-18 interact with the expected group i intron cognates of the aminoacyl-acceptor st ...200211854463
mutants of the carotene cyclase domain of al-2 from neurospora crassa.phytoene synthase and carotene cyclase, two key enzymes in carotenoid biosynthesis, are encoded by two separate genes in bacteria and plants, but by a single bifunctional gene in fungi. the cyclase function has been demonstrated for the products of the genes crtyb from the basidiomycete xanthophyllomyces dendrorhous, and for carra and carrp from the zygomycetes phycomyces blakesleeanus and mucor circinelloides, respectively. these three genes are highly similar to al-2 from neurospora crassa. ta ...200211862485
identification and characterization of the genes encoding the core histones and histone variants of neurospora crassa.we have identified and characterized the complete complement of genes encoding the core histones of neurospora crassa. in addition to the previously identified pair of genes that encode histones h3 and h4 (hh3 and hh4-1), we identified a second histone h4 gene (hh4-2), a divergently transcribed pair of genes that encode h2a and h2b (hh2a and hh2b), a homolog of the f/z family of h2a variants (hh2az), a homolog of the h3 variant cse4 from saccharomyces cerevisiae (hh3v), and a highly diverged h4 ...200211901114
the fle1 gene encoding a c2h2 zinc finger protein co-ordinates male and female sexual differentiation in podospora anserina.the flexuosa (fle1-1) mutant, isolated in podospora anserina, displays vegetative defects and two antagonistic sexual phenotypes: it produces several 1000-fold fewer microconidia (male gametes) than the wild-type strain and, conversely, more abundant protoperithecia (female organs). cloning and sequencing of the fle1 gene and of cdna identified an open reading frame encoding a 382-amino-acid polypeptide with two c2h2 zinc finger motifs. the predicted fle1 protein shares 46% identity with the flb ...200211918811
evolution of the fot1 transposons in the genus fusarium: discontinuous distribution and epigenetic understand the evolution of fot1, a member of the pogo family widely dispersed in ascomycetes, we have performed a phylogenetic survey across the genus fusarium divided into six sections. the taxonomic distribution of fot1 is not homogeneous but patchy; it is prevalent in the fusarium oxysporum complex, absent in closely related sections, and found in five species from the most distant section martiella. multiple copies of fot1 were sequenced from each strain in which the element occurs. in t ...200211919292
the dna damage response in filamentous fungi.the mechanisms used by fungal cells to repair dna damage have been subjects of intensive investigation for almost 50 years. as a result, the model yeasts schizosaccharomyces pombe and saccharomyces cerevisiae have led the way in yielding critical insights into the nature of the dna damage response. at the same time, largely through the efforts of etta kafer, hirokazu inoue, and colleagues, a substantial collection of aspergillus nidulans and neurospora crassa dna repair mutants has been identifi ...200211929209
a genetic map of blumeria graminis based on functional genes, avirulence genes, and molecular markers.a genetic map of the powdery mildew fungus, blumeria graminis f. sp. hordei, an obligate biotrophic pathogen of barley, is presented. the linkage analysis was conducted on 81 segregating haploid progeny isolates from a cross between 2 isolates differing in seven avirulence genes. a total of 359 loci were mapped, comprising 182 amplified fragment length polymorphism markers, 168 restriction fragment length polymorphism markers including 42 ltr-retrotransposon loci and 99 expressed sequence tags ( ...200211929213
functional analysis of the escherichia coli molybdopterin cofactor biosynthesis protein moea by site-directed mutagenesis.five moea mutants were generated by replacing some conserved amino acids of moea by site-directed mutagenesis. the mutants were assayed for the ability to restore in vivo nitrate reductase activity of the moea mutant escherichia coli jrg97 and in vitro neurospora crassa nit-1 nitrate reductase activity. the replacements asp59alagly60ala, asp259ala, pro298alapro301ala abolished the function of moea in mo-molybdopterin formation and stabilization, reflected in the inability to restore nitrate redu ...200211934270
cia30 complex i assembly factor: a candidate for human complex i deficiency?the human mitochondrial nadh:ubiquinone oxidoreductase (complex i), the first complex of the oxidative phosphorylation system, is composed of at least 42 subunits. little is known about the assembly process of these subunits into the mature complex. recently, two proteins in neurospora crassa have been found to be involved in the assembly of complex i. these proteins are not constituent parts of the mature complex but are associated with smaller intermediate complexes of the assembly process and ...200211935339
genes encoding chimeras of neurospora crassa erg-3 and human tm7sf2 proteins fail to complement neurospora and yeast sterol c-14 reductase mutants.the human gene tm7sf2 encodes a polypeptide (sr-1) with high sequence similarity to sterol c-14 reductase, a key sterol biosynthetic enzyme in fungi, plants and mammals. in neurospora and yeast this enzyme is encoded by the erg-3 and erg24 genes respectively. in an effort to demonstrate sterol c-14 reductase activity for sr-1 we constructed six recombinant genes coding for chimeras of the neurospora erg-3 and sr-1 protein sequences and tested them for complementation of the neurospora erg-3 muta ...200211937680
effects of radiation during space flight on microorganisms and plants on the biosatellite ii and gemini xi missions.the results of recent experiments with the lysogenic bacteria, escherichia coli and salmonella typhimurium, the bread mold neurospora crassa and the flowering plant tradescantia on the biosatellite ii and gemini xi missions will be summarized. in the lysogenic bacteria experiment (dr. rudolf h.t. mattoni, nus corporation) on the biosatellite ii mission significant effects of space flight were found on both growth rate and the induction of prophage. in that part of the neurospora experiment on bo ...196911949688
interaction of mitochondria with microtubules in the filamentous fungus neurospora crassa.the establishment and maintenance of the 3d structure of eukaryotic cells depends on active transport and positioning of organelles along cytoskeletal elements. the biochemical basis of these processes is only poorly understood. we analysed the interaction of mitochondria with microtubules in the filamentous fungus neurospora crassa. mitochondria were fluorescently labelled by expression of matrix-targeted green fluorescent protein. upon isolation, mitochondria collapsed to round spherical struc ...200211956324
molecular and biochemical characterization of the neurospora crassa glycogen synthase encoded by the gsn cdna.glycogen synthases catalyze the transfer of a glucosyl moiety from a nucleotide phosphosugar to a nascent glycogen chain via an alpha1-->4 linkage. although many genes coding for glycogen synthases have been described, the enzymes from rabbit and yeast are the best characterized. the fungus neurospora crassa accumulates glycogen during exponential growth, and mobilizes it at the onset of stationary phase, or when placed at high temperature or starved for carbon. through a pcr methodology, the gs ...200211976968
cloning and characterization of vmaa, the gene encoding a 69-kda catalytic subunit of the vacuolar h+-atpase during alkaline ph mediated growth of aspergillus oryzae.screening of a cdna library constructed under alkaline ph mediated growth of aspergillus oryzae implicated a vacuolar h+-atpase gene (vmaa) as a putative candidate involved in alkaline ph adaptation. a. oryzae vmaa genomic dna extended to 2072 bp including three introns and encoded a protein of 605 amino acids. vmaap was homologous to vma-1p from neurospora crassa (71%), vma1p from saccharomyces cerevisiae (69%) and atp6a2 from human (49%). the vmaa cdna complemented s. cerevisiae v-atpase disru ...200212007818
[arg-13 is implicated in mitochondrial ornithine transport in neurospora crassa].arg-13 is a leaky mutation involved in arginine metabolism. a tight selection is developed using similar amount of lysine and ornithine replacing other nitrogen source in minimal medium. this selection strongly inhibits the growth of arg-13 under stringent sorbose/glucose condition but allows arg-13 to grow under spot test conditions. as ornithine is build up through mitochondrial ornithine biosynthesis and transport from cytoplasm to mitochondria, arg-13 is combined in genetic crosses with arg- ...199812016957
carnitine biosynthesis in neurospora crassa: identification of a cdna coding for epsilon-n-trimethyllysine hydroxylase and its functional expression in saccharomyces cerevisiae.the biosynthesis of l-carnitine in eukaryotic organisms was first elucidated in the ascomycete neurospora crassa. the first step of the pathway is catalysed by epsilon-n-trimethyllysine hydroxylase (tmlh), which converts epsilon-n-trimethyllysine into beta-hydroxy-n-epsilon-trimethyllysine in a reaction dependent on alpha-ketoglutarate, fe2+ and oxygen. here we report on the cloning of the n. crassa tmlh cdna and its functional expression in saccharomyces cerevisiae. the tmlh cdna contains an op ...200212023072
a 3d model of the voltage-dependent anion channel (vdac).eukaryotic porins are a group of membrane proteins whose best known role is to form an aqueous pore channel in the mitochondrial outer membrane. as opposed to the bacterial porins (a large family of protein whose 3d structure has been determined by x-ray diffraction), the structure of eukaryotic porins (also termed vdacs, voltage-dependent anion-selective channels) is still a matter of debate. we analysed the secondary structure of vdac from the yeast saccharomyces cerevisiae, the fungus neurosp ...200212044860
identification, cloning and expression of the mouse n-acetylglutamate synthase ureotelic animals, n-acetylglutamate (nag) is an essential allosteric activator of carbamylphosphate synthetase i (cpsi), the first enzyme in the urea cycle. nag synthase (nags; ec catalyses the formation of nag from glutamate and acetyl-coa in liver and intestinal mitochondria. this enzyme is supposed to regulate ureagenesis by producing variable amounts of nag, thus modulating cpsi activity. moreover, inherited deficiencies in nags have been associated with hyperammonaemia, probabl ...200212049647
disruption of iron-sulphur cluster n2 from nadh: ubiquinone oxidoreductase by site-directed mutagenesis.we have cloned and inactivated, by repeat-induced point mutations, the nuclear gene encoding the 19.3 kda subunit of complex i (ec from neurospora crassa, the homologue of the bovine psst polypeptide. mitochondria from mutant nuo19.3 lack the peripheral arm of complex i while its membrane arm accumulates. transformation with wild-type cdna rescues this phenotype and assembly of complex i is restored. to interfere with assembly of a proposed bound iron-sulphur cluster, site-directed muta ...200212049648
toeprint analysis of the positioning of translation apparatus components at initiation and termination codons of fungal mrnas.the ability to map the position of ribosomes and their associated factors on mrnas is critical for an understanding of translation mechanisms. earlier approaches to monitoring these important cellular events characterized nucleotide sequences rendered nuclease-resistant by ribosome binding. while these approaches furthered our understanding of translation initiation and ribosome pausing, the pertinent techniques were technically challenging and not widely applied. here we describe an alternative ...200212054887
a cytosine methyltransferase homologue is essential for repeat-induced point mutation in neurospora crassa.during sexual development, neurospora crassa inactivates genes in duplicated dna segments by a hypermutation process, repeat-induced point mutation (rip). rip introduces c:g to t:a transition mutations and creates targets for subsequent dna methylation in vegetative tissue. the mechanism of rip and its relationship to dna methylation are not fully understood. mutations in dim-2, a dna methyltransferase (dmt) responsible for all known cytosine methylation in neurospora, does not prevent rip. we u ...200212072568
the upr-1 gene encodes a catalytic subunit of the dna polymerase zeta which is involved in damage-induced mutagenesis in neurospora crassa.the upr-1 mutant was one of the first mutagen-sensitive mutants to be isolated in neurospora crassa. however, the function of the upr-1 gene has not yet been elucidated, although some genetic and biochemical data have been accumulated. in order to clone the upr-1 gene, we performed a chromosome walk from the mat locus, the closest genetic marker to upr-1 for which a molecular probe was available, towards the centromere, and a chromosomal contig of about 300-400 kb was constructed. some of these ...200212073042
functional analysis of the c6 zinc finger gene pro1 involved in fungal sexual development.the pro1 gene, controlling fruiting body development in the homothallic ascomycete sordaria macrospora, encodes a c6 zinc finger protein with a typical dna binding domain of gal4-like c6 zinc finger proteins as well as a putative nuclear targeting signal. in the corresponding mutant pro1, the pro1 gene is deleted, and the transition of primordia into mature fruiting bodies is prevented. to further characterize the pro1 polypeptide, the yeast system was used for identifying a transactivation doma ...200212081464
interaction between mating-type proteins from the homothallic fungus sordaria macrospora.mating-type genes control sexual development in ascomycetes. little is known about their function in homothallic species, which are self-fertile and do not require a mating partner for sexual reproduction. the function of mating-type genes in the homothallic fungus sordaria macrospora was assayed using a yeast system in order to find properties typical of eukaryotic transcription factors. we were able to demonstrate that the mating-type proteins smta-1 and smta-1 have domains capable of activati ...200212111096
a regulator gene for acetate utilisation from neurospora crassa.the neurospora crassa homologue of the aspergillus nidulans regulatory gene facb has been cloned. the gene encodes a putative transcriptional activator of 865 amino acids that contains a dna-binding domain with a zn(ii)(2)cys(6) binuclear cluster, a linker region and a leucine zipper-like heptad repeat. two internal amino acid sequences are identical to peptide sequences determined from proteolytic fragments of a dna-binding protein complex specific for genes involved in acetate utilisation and ...200212111557
an osmosensing histidine kinase mediates dicarboximide fungicide resistance in botryotinia fuckeliana (botrytis cinerea).a two-component histidine protein kinase gene, homologous to os-1 from neurospora crassa, was cloned and sequenced from a single ascospore isolate of botryotinia fuckeliana. a series of nine spontaneous mutants resistant to dicarboximide fungicides was selected from this strain and characterized with respect to fungicide resistance and osmotic sensitivity. genetic crosses of the mutants with an authentic daf1 strain showed that the phenotypes mapped to this locus. single point mutations (seven t ...200212135574
cloning and characterization of the gene encoding ribosomal p0 phosphoprotein from neurospora crassa.a gene for ribosomal protein p0 that belongs to the family of ribosomal p proteins was isolated from a neurospora crassa cdna library, using polyclonal antibodies against recombinant p0 protein from saccharomyces cerevisiae. this is the first gene for ribosomal p0 protein to be cloned from filamentous fungi. the derived p0 protein sequence has a strong homology to other eukaryotic p0 proteins; yet, there is a notable alteration in the conservative c-terminal region, placing this protein among th ...200212136931
cellular responses to environmental salinity in the halophilic black yeast hortaea werneckii.the development of crop plants with increased salt tolerance necessitates the study of naturally salt-tolerant eukaryotic species. we studied the bio-synthesis of glycerol as a compatible solute in the halophilic eukaryotic microorganism, black yeast hortaea werneckii. a restriction fragment-differential display technique was used to investigate the transcriptome of the organism. eight differentially expressed genes were identified in response to growth at different salinities. although the puta ...200212139614
regulation and oxidation of two large monofunctional catalases.the two neurospora crassa catalase genes cat-1 and cat-3 were shown to encode cat-1 and cat-3 large monofunctional catalases. cat-1 and cat-3 genes are regulated differentially during the asexual life cycle and under stress conditions. a stepwise increase in catalase activity occurs during conidiation. conidia have 60 times more catalase activity than exponentially growing hyphae. cat-1 activity was predominant in conidia, during germination and early exponential growth. it was induced during pr ...200212160934
cloning and expression of an endo-1,6-beta-d-glucanase gene (neg1) from neurospora crassa.a gene (neg1) encoding an endo-1,6-beta-d-glucanase from neurospora crassa was cloned. the putative neg1 was 1443-bp long and encoded a mature endo-1,6-beta-d-glucanase protein of 463 amino acids and signal peptide of 17 amino acids. the purified recombinant protein (neg1) obtained from escherichia coli showed 1,6-beta-d-glucanase activity. no genes similar in sequence were found in yeasts and fungi.200212162562
[effect of microbial elicitors on sclerotia biomass and carotenoid yield in surface cultures of penicillium sp.pt95].six kinds of elicitors were prepared respectively from neurospora crassa, monascus purpureus, sporobolomyces roseus, rhodotorula rubra, nocardia sp. n89 and actinoplanes sp. a05. when penicillium sp. pt95 was incubated in czapek's agar plates containing appropriate amounts of elicitors, both its sclerotia biomass and carotenoid content accumulated in sclerotia were enhanced significantly (p < 0.01). among tested elicitors, the elicitors from the fungi n. crassa, m. purpureus, s.-roseus and r. ru ...200212192877
repeat-induced point mutation (rip) in magnaporthe grisea: implications for its sexual cycle in the natural field context.repeat-induced point mutation (rip) is a process that detects dna duplications and peppers their sequences with c:g to t:a transitions in the sexual phase of the life cycle. so far, this unique mechanism has been identified as a currently active process in only two fungal species, neurospora crassa and podospora anserina. to determine whether a rip-like process operates in the plant pathogenic fungus magnaporthe grisea, the retrotransposon maggy and the hygromycin b phosphotransferase gene were ...200212207702
improving the efficiency of gene replacements in neurospora crassa: a first step towards a large-scale functional genomics, we report the use of the mating type heterokaryon incompatibility system as a counterselection to increase the probability of identifying gene replacements in neurospora crassa. we compared the frequencies of gene replacements observed among transformants obtained by using plasmids with or without the mat a-1(+) gene (hereby called "toxic gene") placed adjacent to disruption cassettes. on an average, we were 20x more likely to identify a correct gene replacement by incorporating the toxic ...200212223190
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
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
the hypocrea jecorina gal10 (uridine 5'-diphosphate-glucose 4-epimerase-encoding) gene differs from yeast homologues in structure, genomic organization and 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
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
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
acyl derivatives of homoserine as substrates for homocysteine synthesis in neurospora crassa, yeast, and escherichia coli. 196712325384
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
identification and functional expression of taha, a filamentous fungal gene involved in copper trafficking to the secretory pathway in trametes 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
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
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
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
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
isolation and genetic characterization of the neurospora crassa rev1 and rev7 homologs: evidence for involvement in damage-induced 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
Displaying items 1301 - 1400 of 5889