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the isolation and characterization of nrc-1 and nrc-2, two genes encoding protein kinases that control growth and development in neurospora crassa.using an insertional mutagenesis approach, a series of neurospora crassa mutants affected in the ability to control entry into the conidiation developmental program were isolated. one such mutant, gth16-t4, was found to lack normal vegetative hyphae and to undergo constitutive conidiation. the affected gene has been named nrc-1, for nonrepressible conidiation gene #1. the nrc-1 gene was cloned from the mutant genomic dna by plasmid rescue, and was found to encode a protein closely related to the ...19989584090
role of the negative charges in the cytosolic domain of tom22 in the import of precursor proteins into mitochondria.tom22 is an essential mitochondrial outer membrane protein required for the import of precursor proteins into the organelles. the amino-terminal 84 amino acids of tom22 extend into the cytosol and include 19 negatively and 6 positively charged residues. this region of the protein is thought to interact with positively charged presequences on mitochondrial preproteins, presumably via electrostatic interactions. we constructed a series of mutant derivatives of tom22 in which 2 to 15 of the negativ ...19989584158
cell wall active antifungal compounds produced by the marine fungus hypoxylon oceanicum ll-15g256. iii. biological properties of 15g256 gamma.15g256 gamma is a cyclic lipopeptide antifungal agent discovered in a mechanism of action screen for cell wall acting antifungal agents. the compound shows moderate activity in both greenhouse tests against plant disease caused by pathogenic fungi and in in vitro tests against human fungal pathogens. microscopic examination of treated fungi suggests that the compound acts by the inhibition of cell wall biosynthesis. however, in vitro inhibition of neurospora crassa glucan and chitin synthase wer ...19989589067
bacterial expression and characterization of the mitochondrial outer membrane channel. effects of n-terminal modifications.several forms of the voltage-dependent anion-selective channel (vdac) have been expressed at high yield in escherichia coli. full-length constructs of the proteins of neurospora crassa and saccharomyces cerevisiae (ncvdac and scvdac) have been made with 20-residue-long, thrombin-cleavable, his6-containing n-terminal extensions. ncvdac purified from bacteria or mitochondria displays a far-uv cd spectrum (in 1% lauryl dimethylamine oxide at ph 6-8) similar to that of bacterial porins, indicating e ...19989593723
complex i from the fungus neurospora crassa.respiratory chain complex i is a complicated enzyme of mitochondria, that couples electron transfer from nadh to ubiquinone to the proton translocation across the inner membrane of the organelle. the fungus neurospora crassa has been used as one of the main model organisms to study this enzyme. complex i is composed of multiple polypeptide subunits of dual genetic origin and contains several prosthetic groups involved in its activity. most subunits have been cloned and those binding redox centre ...19989593837
transcriptional activation of the aspergillus nidulans gpda promoter by osmotic signals.a differentially expressed gpda cdna clone was isolated from nacl-adapted aspergillus nidulans (fgsc359) and identified as glyceraldehyde-3-phosphate dehydrogenase (gpda) on the basis of its nucleotide sequence. the level of gpda rna substantially increased in cultures gradually adapted to nacl but was greatly reduced in cultures exposed briefly to a high concentration of nacl. a pyrg auxotroph of a. nidulans (a773) was cotransformed with a gpda-uida construct and a plasmid containing the neuros ...19989603839
cloning and sequencing of a cdna encoding a copper-zinc superoxide dismutase enzyme from the marine yeast debaryomyces hansenii.cu-zn superoxide dismutase (sod-1) is a ubiquitously occurring eukaryotic enzyme with a variety of important effects on respiring organisms. a gene (dhsod-1) encoding a cu-zn superoxide dismutase of the marine yeast debaryomyces hansenii was cloned using mrna by the rt-pcr technique. the deduced amino-acid sequence shows approximately 70% homology with that of cytosolic superoxide dismutase from saccharomyces cerevisiae and neurospora crassa, as well as lower homologies (between 55 and 65%) with ...19989605507
molecular characterisation of the 76 kda iron-sulphur protein subunit of potato mitochondrial complex i.genes encoding subunits of complex i (ec 1.6.5.3) of the mitochondrial respiratory chain vary in their locations between the mitochondrial and nuclear genomes in different organisms, whereas genes for a homologous multisubunit complex in chloroplasts have to date only been found on the plastid genome. in potato (solanum tuberosum l.), the gene coding for the mitochondrial 76 kda iron-sulphur protein is identified in the nuclear genome. the gene is transcribed into polyadenylated mrna which is mo ...19989615461
cos1, a two-component histidine kinase that is involved in hyphal development in the opportunistic pathogen candida albicans.two-component histidine kinases recently have been found in eukaryotic organisms including fungi, slime molds, and plants. we describe the identification of a gene, cos1, from the opportunistic pathogen candida albicans by using a pcr-based screening strategy. the sequence of cos1 indicates that it encodes a homolog of the histidine kinase nik-1 from the filamentous fungus neurospora crassa. cos1 is also identical to a gene called canik1 identified in c. albicans by low stringency hybridization ...19989618540
analysis of models involving enzymatic activities for the occurrence of c-->t transition mutations during repeat-induced point mutation (rip) in neurospora crassa.the phenomenon of repeat-induced point mutation (rip), acting during the sexual phase of the model eukaryote neurospora crassa, is considered to study the putative in vivo relationships existing between cellular levels of s-adenosylmethionine (sam), cytosine methylation and the occurrence of c-->t transition mutations. we analyse the kinetic behaviour of the different enzymatic models proposed to explain the underlying mutagenic mechanisms of rip. the dependence of the mutation rate on the cellu ...19989628839
interaction between major nitrogen regulatory protein nit2 and pathway-specific regulatory factor nit4 is required for their synergistic activation of gene expression in neurospora crassa.in neurospora crassa, the major nitrogen regulatory protein, nit2, a member of the gata family of transcription factors, controls positively the expression of numerous genes which specify nitrogen catabolic enzymes. expression of the highly regulated structural gene nit-3, which encodes nitrate reductase, is dependent upon a synergistic interaction of nit2 with a pathway-specific control protein, nit4, a member of the gal4 family of fungal regulatory factors. the nit2 and nit4 proteins both bind ...19989632783
the preprotein translocation channel of the outer membrane of mitochondria.the preprotein translocase of the outer membrane of mitochondria (tom complex) facilitates the recognition, insertion, and translocation of nuclear-encoded mitochondrial preproteins. we have purified the tom complex from neurospora crassa and analyzed its composition and functional properties. the tom complex contains a cation-selective high-conductance channel. upon reconstitution into liposomes, it mediates integration of proteins into and translocation across the lipid bilayer. tom complex pa ...19989635430
the sensor regions of vdac are translocated from within the membrane to the surface during the gating processes.the motion of the sensor regions in a mitochondrial voltage-gated channel called vdac were probed by attaching biotin at specific locations and determining its ability to bind to added streptavidin. site-directed mutagenesis was used to introduce single cysteine residues into neurospora crassa vdac (naturally lacks cysteine). these were chemically biotinylated and reconstituted into planar phospholipid membranes. in the 19 sites examined, only two types of results were observed upon streptavidin ...19989635747
functional analysis of different regions of the positive-acting cys3 regulatory protein of neurospora crassa.in the filamentous fungus neurospora crassa during conditions of sulfur limitation, cys3, a major positive-acting regulatory protein, turns on the expression of an entire set of genes which encode permeases and enzymes involved in the acquisition of sulfur from environmental sources. cys3 functions as a homodimeric protein and possesses a b-zip domain that confers sequence-specific dna binding. expression of various hybrid gal4-cys3 fusion proteins in yeast was used to detect regions involved in ...19989644202
dab1: a degenerate retrotransposon-like element from neurospora crassa.a repeated dna sequence in the genome of neurospora crassa has been identified as a family of degenerate retroelements. retroelements encode protein sequences with clear homology to the reverse transcriptase, rnase h and endonuclease products of the pol genes common to retroviruses and retrotransposons. these sequence comparisons place the n. crassa element within the gypsy group of retrotransposons, akin to other elements found in filamentous fungi. however, the neurospora element is defective, ...19989648750
analysis of complex formation between hsp80 and hsp70, cytosolic molecular chaperones of neurospora crassa, by enzyme-linked immunosorbent assays (elisa).a physical association between hsp70 and hsp80, the major cytosolic stress proteins of neurospora crassa, was demonstrated previously by interprotein crosslinking and by binding of hsp80 to hsp70 immobilized on atp-agarose. in the present study, [hsp70: hsp80] complex formation was analyzed by enzyme-linked immunosorbent assays (elisa), using specific antibodies. one protein was fixed onto elisa plate wells and binding of the second mobile protein was monitored by retention of its cognate igg. b ...19989666311
the membrane domain of complex i is not assembled in the stopper mutant e35 of neurospora.the assembly of mitochondrial nadh: ubiquinone oxidoreductase (complex i) was studied in the e35 stopper mutant of neurospora crassa at different times during growth in liquid media. assembly of complex i as well as of its membrane domain is impaired in this strain throughout the growth period. nevertheless, a structure that resembles the peripheral aim of the enzyme is still formed in the mitochondria of this mutant. the absence of the membrane domain of complex i in e35 can be attributed to th ...19989666316
complementation of the mpg1 mutant phenotype in magnaporthe grisea reveals functional relationships between fungal hydrophobins.the functional relationship between fungal hydrophobins was studied by complementation analysis of an mpg1(-) gene disruption mutant in magnaporthe grisea. mpg1 encodes a hydrophobin required for full pathogenicity of the fungus, efficient elaboration of its infection structures and conidial rodlet protein production. seven heterologous hydrophobin genes were selected which play distinct roles in conidiogenesis, fruit body development, aerial hyphae formation and infection structure elaboration ...19989670001
pleiotropic deficiencies of the laccase-derepressed mutant lah-1 are caused by constitutively increased expression of the cross-pathway control gene cpc-1 in neurospora crassa.in neurospora crassa, expression of the laccase gene is induced by treatment with the protein synthesis inhibitor cycloheximide (chx). this expression is mediated by cpc1, which acts as a general transcriptional activator when mycelia are treated with chx or starved for any one of the amino acids. a laccase-derepressed mutant, lah-1, shows pleiotropic deficiencies in growth, hyphal morphology, chx sensitivity, and production of protoperithecia. moreover, in the lah-1 mutant, transcript levels of ...19989671030
identification, cloning and characterization of a derepressible na+-coupled phosphate transporter in saccharomyces cerevisiae.based on the high sequence homology between the yeast orf ybr296c (accession number p38361 in the swiss-prot database) and the pho4 gene of neurospora crassa, which codes for a na+/pi cotransporter with twelve putative transmembrane domains, the ybr296c orf was considered to be a promising candidate gene for a plasma membrane-bound phosphate transporter in saccharomyces cerevisiae. therefore, this gene, here designated pho89, was cloned and a set of deletion mutants was constructed. we then stud ...19989671031
calcineurin subunit b is required for normal vegetative growth in neurospora crassa.a series of neurospora crassa mutants affected in the ability to regulate entry into conidiation (an asexual developmental program) were isolated by using an insertional mutagenesis procedure followed by a screening protocol. one of the mutants isolated by this approach consisted entirely of cells with an abnormal morphology. the mutant produces chains of swollen septated cells. the developmentally regulated ccg-1 gene is constitutively expressed in these cells, suggesting that they have entered ...19989680955
analysis of two transcription activation elements in the promoter of the developmentally regulated con-10 gene of neurospora crassa.the con-10 gene of neurospora crassa is activated during conidiation. crs-b (conidiation response sequence-b) and cge (con-10 general enhancer) elements have been proposed to function as sites of transcriptional activation and as possible elements that confer developmental regulation to con-10. in a specific analysis of the roles of these elements we found that two crs-b elements are necessary for full activation of con-10 during macroconidiation, whereas two cge elements are functionally redund ...19989680956
tissue-specific repression of starvation and stress responses of the neurospora crassa con-10 gene is mediated by rco1.the neurospora crassa con-10 gene is weakly expressed in mycelia but is induced approximately 1000-fold during macroconidiation. studies of the promoter elements and trans-acting factors that regulate con-10 expression are needed to gain a detailed understanding of developmental regulation. the rco-1 mutant displays a 10-fold elevated basal level of expression of con-10. in contrast to the wild type, con-10 expression in mycelia of the rco-1 mutant was rapidly induced to high levels by starvatio ...19989680957
the essential aspergillus nidulans gene pmaa encodes an homologue of fungal plasma membrane h(+)-atpases.pmaa, an aspergillus nidulans gene encoding a p-atpase, has been cloned by heterologous hybridization with the yeast pma1 gene. the putative 990-residue pmaa polypeptide shows 50% identity to saccharomyces cerevisiae and neurospora crassa plasma membrane h(+)-atpases and weak (19-26%) identity to other yeast p-type cation-translocating atpases. pmaa contains all catalytic domains characterizing h(+)-atpases. pmaa transcript levels are not regulated by pacc, the transcription factor mediating ph ...19989680959
photoregulation of cot-1, a kinase-encoding gene involved in hyphal growth in neurospora crassa.blue light plays a key role as an environmental signal in the regulation of growth and development of fungi and plants. here we demonstrate that in neurospora crassa hyphae branch more frequently in cultures grown in light. previous studies had identified cot-1 as a gene that controls apical hyphal cell elongation. in the cot-1 mutant, cessation of elongation is accompanied by hyperbranching. here we demonstrate that the cot-1 gene encodes two transcript species of about 2100 nt (cot-1 (s)) and ...19989680960
heat shock effects on second messenger systems of neurospora crassa.exposure of growing hyphae of neurospora crassa to heat shock (44 degrees c) or ethanol (2.6 m) for 1 h induced a significant increase in the camp level, which reached a maximum approximately 2 min after the beginning of treatment and then decreased to control values despite continued heat or ethanol exposure. a 10-s heat shock or a 5-s ethanol shock also resulted in a transient camp increase 2 min after the pulse. heat shock or ethanol treatment led to an increase in the amount of catalytic sub ...19989683659
analysis of conversion tracts associated with recombination events at the am locus of neurospora crassa.in cross b163, heteroallelic am1 am6 and heterozygous for both conventional genetic flanking markers and closer molecular markers, we previously found that the majority on flanker exchanges were remote from events that generated prototrophic recombinants. we report here that natural polymorphisms distinguishing the parents of cross b163 also include sequences within and closely flanking am. segregation of these markers in b163 prototrophs confirms that the majority of meiotic recombination event ...19989683674
cpc-3, the neurospora crassa homologue of yeast gcn2, encodes a polypeptide with juxtaposed eif2alpha kinase and histidyl-trna synthetase-related domains required for general amino acid control.based on characteristic amino acid sequences of kinases that phosphorylate the alpha subunit of eukaryotic translation initiation factor 2 (eif2alpha kinases), degenerate oligonucleotide primers were constructed and used to polymerase chain reaction-amplify from genomic dna of neurospora crassa a sequence encoding part of a putative protein kinase. with this sequence an open reading frame was identified encoding a predicted polypeptide with juxtaposed eif2alpha kinase and histidyl-trna synthetas ...19989685394
trichostatin a causes selective loss of dna methylation in neurospora.both dna methylation and hypoacetylation of core histones are frequently associated with repression of gene expression. possible connections between these processes were investigated by taking advantage of genes controlled by methylation in neurospora crassa. trichostatin a (tsa), a potent inhibitor of histone deacetylase, derepressed a copy of hph that was repressed by dna methylation which resulted from repeat-induced point mutation (rip) acting on sequences flanking hph. derepression by tsa w ...19989689097
a methylated neurospora 5s rrna pseudogene contains a transposable element inactivated by repeat-induced point mutation.in an analysis of 22 of the roughly 100 dispersed 5s rrna genes in neurospora crassa, a methylated 5s rrna pseudogene, psi63, was identified. we characterized the psi63 region to better understand the control and function of dna methylation. the 120-bp 5s rrna-like region of psi63 is interrupted by a 1.9-kb insertion that has characteristics of sequences that have been modified by repeat-induced point mutation (rip). we found sequences related to this insertion in wild-type strains of n. crassa ...19989691037
analysis of a distal cluster of binding elements and other unusual features of the promoter of the highly regulated nit-3 gene of neurospora crassa.in neurospora crassa, the expression of the nit-3 gene (nitrate reductase) is dependent upon nitrogen derepression and nitrate induction and is regulated by two positive-acting transcription factors, nit2 and nit4, and a negative regulator, nmr. the presence of a tightly linked cluster of nit2 and nit4 binding sites suggested that their close spacing might be required for a synergistic interaction of the nit2 and nit4 proteins. we show here that the nit2 and nit4 binding sites can be separated w ...19989693010
isolation and characterization of strains defective in vacuolar ornithine permease in neurospora crassa.vacuolar uptake of ornithine and lysine was characterized in neurospora crassa using a cupric ion permeabilization system. michaelis constants were measured as 1.4 mm for lysine and 11.0 mm for ornithine, and maximal velocities were determined. vacuolar lysine uptake was shown to be inhibited competitively by l-arginine and histidine while ornithine uptake was inhibited by a variety of amino acids. strains defective in the vacuolar ornithine permease were isolated using a filtration enrichment m ...19989693025
photoregulation of cot-1, a kinase-encoding gene involved in hyphal growth in neurospora crassa.blue light plays a key role as an environmental signal in the regulation of growth and development of fungi and plants. here we demonstrate that in neurospora crassa hyphae branch more frequently in cultures grown in light. previous studies had identified cot-1 as a gene that controls apical hyphal cell elongation. in the cot-1 mutant, cessation of elongation is accompanied by hyperbranching. here we demonstrate that the cot-1 gene encodes two transcript species of about 2100 nt (cot-1 (s)) and ...19989693026
phosphate permeases of saccharomyces cerevisiae.the pho84 and pho89 genes of saccharomyces cerevisiae encode two high-affinity phosphate cotransporters of the plasma membrane. hydropathy analysis suggests a secondary structure arrangements of the proteins in 12 transmembrane domains. the derepressible pho84 and pho89 transporters appear to have characteristic similarities with the phosphate transporters of neurospora crassa. the pho84 protein catalyzes a proton-coupled phosphate transport at acidic ph, while the pho89 protein catalyzes a sodi ...19989693717
redox components and structure of the respiratory nadh:ubiquinone oxidoreductase (complex i).the proton-pumping nadh:ubiquinone oxidoreductase is the first complex in the respiratory chains of many purple bacteria and of mitochondria of most eucaryotes. the bacterial complex consists of 14 different subunits. the mitochondrial complex contains at least 29 additional proteins that do not directly participate in electron transfer and proton translocation. we analysed electron micrographs of isolated and negatively stained complex i particles from escherichia coli and neurospora crassa and ...19989693737
a novel pathway for cytochromes c biogenesis in chloroplasts.the cytochromes c are a useful model for the study of the pathways and mechanisms of assembly of the cofactor-containing components of energy transducing membranes. genetic analyses have identified proteins that are required for the assembly of c-type cytochromes in mitochondria, bacteria and chloroplasts. the components of the pathway operating in fungal and animal mitochondria, i.e. the cytochrome (cyt) c and c1 heme lyases in the intermembrane space, were identified over a decade ago through ...19989693743
changes in fatty acid composition of neurospora crassa accompany sexual development and ascospore germination.fatty acid composition was determined during several stages of sexual development in neurospora crassa. triacylglycerol was the predominant acyl lipid in cultures undergoing sexual development. the absolute amounts of triacylglycerol in fertilized cultures varied over time, in contrast to control (unfertilized or mock-fertilized) cultures, in which the amount of triacylglycerol decreased linearly with age. in cultures competent to undergo sexual development, alpha-linoleate was the predominant f ...19989695905
distribution patterns of 104 kda stress-associated protein in rice.a 104 kda protein (sap 104) accumulates in rice seedlings in response to several abiotic stress conditions and immunological homologues of rice sap 104 have been detected in several monocot and dicot species, as also neurospora crassa, a fungus. in this report, we show that the amino acid sequence of a tryptic peptide generated from purified sap 104 bears significant homology with an atp-binding domain of the hsp 100 family proteins of arabidopsis thaliana and glycine max. it is further shown th ...19989700064
apolar growth of neurospora crassa leads to increased secretion of extracellular proteins.protein secretion in filamentous fungi has been shown to be restricted to actively growing hyphal tips. to determine whether an increase in the amount f growing surface area of a fungus can lead to an increase in the amount of protein secretion, we examined secretion in a temperature-sensitive neurospora crassa mcb mutant that shows a loss of growth polarity when incubated at restrictive temperature. incubation of the mcb mutant at restrictive temperature results in a three- to fivefold increase ...19989701815
the chlamydomonas reinhardtii moco carrier protein is multimeric and stabilizes molybdopterin cofactor in a molybdate charged form.in chlamydomonas reinhardtii, molybdopterin cofactor (moco) able to reconstitute active nitrate reductase (nr) with apoenzyme from the neurospora crassa mutant nit-1 was found mostly bound to a carrier protein (cp). this protein is scarce in the algal free extracts and has been purified 520-fold. mococp is a protein of 64 kda with subunits of 16.5 kda and an isoelectric point of 4.5. in contrast to free moco, moco bound to cp was remarkably protected against inactivation under both aerobic condi ...19989708903
dynamics of the tom complex of mitochondria during binding and translocation of preproteins.translocation of preproteins across the mitochondrial outer membrane is mediated by the tom complex. this complex consists of receptor components for the initial contact with preproteins at the mitochondrial surface and membrane-embedded proteins which promote transport and form the translocation pore. in order to understand the interplay between the translocating preprotein and the constituents of the tom complex, we analyzed the dynamics of the tom complex of neurospora crassa and saccharomyce ...19989710610
structure of the chromosome vii centromere region in neurospora crassa: degenerate transposons and simple repeats.dna from the centromere region of linkage group (lg) vii of neurospora crassa was cloned previously from a yeast artificial chromosome library and was found to be atypical of neurospora dna in both composition (at rich) and complexity (repetitive). we have determined the dna sequence of a small portion (approximately 16.1 kb) of this region and have identified a cluster of three new retrotransposon-like elements as well as degenerate fragments from the 3' end of tad, a previously identified line ...19989710630
choline depletion, frq mutations, and temperature compensation of the circadian rhythm in neurospora crassa.in the fungus neurospora crassa, the chol-1 mutation blocks the synthesis of the lipid phosphatidylcholine and also lengthens the period of the circadian rhythm of conidiation under conditions of choline depletion. the frq mutations, which have no known metabolic defect, affect both the period of the rhythm and temperature compensation. in this article, the chol-1 mutant strain has been further characterized with respect to its temperature compensation and its interactions with frq. choline depl ...19989711502
structure, function, regulation: experimental analysis in groups of non-excitable cells coupled via permeable junctions.this presentation reviews the functional properties of different permeable junctions (microplasmodesmata of filamentous cyanobacteria, septal pores of fungi, and gap junctions of animals). examples of the distribution of power by ionic fluxes and electric current through permeable cell-cell junctions are considered. based on the analysis of intercellular communication occurring during apical growth of hyphae of wild-type and mutant lines of the mycelial fungus neurospora crassa, approaches to st ...19989718577
in vitro studies on mode of action of antifungal 8.o.4'-neolignans occurring in certain species of virola and related genera of myristicaceae.neutral racemic antifungal alcohols of 8.o.4'-neolignan type, were evaluated for inhibitory activity towards the fungal cell wall, using the whole cell neurospora crassa hyphal growth inhibition assay. results strongly suggested that these compounds could act by inhibiting cell wall polymer synthesis or assembly. active compounds were tested for their inhibitory activities against (1,3)-beta-glucan synthase, an enzyme that catalyzes the synthesis of the major wall polymer (1,3)-beta-glucan. alth ...19989720609
pyrrolnitrin from burkholderia cepacia: antibiotic activity against fungi and novel activities against streptomycetes.a bacterial strain identified as burkholderia cepacia nb-1 was isolated from water ponds in the botanical garden in tübingen, germany, and was found to produce a broad spectrum phenylpyrrole antimicrobial substance active against filamentous fungi, yeasts and gram-positive bacteria. in batch culture containing glycerol and l-glutamic acid, the isolate nb-1 produced the antibiotic optimally late in the growth phase and accumulated a main portion in their cells. isolation and purification of the a ...19989721657
roles in dimerization and blue light photoresponse of the pas and lov domains of neurospora crassa white collar proteins.the genes coding for white collar-1 and white collar-2 (wc-1 and wc-2) have been isolated previously, and their products characterized as zn-finger transcription factors involved in the control of blue light-induced genes. here, we show that the pas dimerization domains present in both proteins enable the wc-1 and wc-2 proteins to dimerize in vitro. homodimers and heterodimers are formed between the white collar (wc) proteins. a computer analysis of wc-1 reveals a second domain, called lov, also ...19989723912
regulation of pectic enzymes from the exo-1 mutant strain of neurospora crassa: effects of glucose, galactose, and galacturonic acid.the exo-1 mutant of neurospora crassa produced and secreted pectolytic activities when incubated in the presence of pectin-containing biological materials. this study shows that polygalacturonase, pectate lyase and pectin lyase activities were induced in media supplemented with galactose or galacturonic acid, indicating that these sugars induced the synthesis of pectinases. pectinesterase activity was undetectable. polygalacturonase activity was better induced by galactose than by galacturonic a ...19989726123
the alkaline single cell gel electrophoresis: a new test for assessing dna single strand breaks in neurospora crassa.the single cell gel electrophoresis (comet assay) is a potent technique in testing double and single strand breaks in dna. in this paper, we present an application of alkaline comet assay to filamentous fungi for genotoxicological assessment of heavy metals for the first time. a wild strain of neurospora crassa sla 4200 was grown in presence of cadmium sulfate (cdso4) (10 microm and 100 microm) for 12 h. protoplasts from 12-h old mycelia were prepared by using novozym 234 and dna damage was eval ...19989729255
the topology of vdac as probed by biotin modification.the outer membrane of mitochondria contains channels called vdac (mitochondrial porin), which are formed by a single 30-kda protein. cysteine residues introduced by site-directed mutagenesis at sites throughout neurospora crassa vdac (naturally devoid of cysteine) were specifically biotinylated prior to reconstitution into planar phospholipid membranes. from previous studies, binding of streptavidin to single biotinylated sites results in one of two effects: reduced single-channel conductance wi ...19989733730
mutations in the dim-1 gene of neurospora crassa reduce the level of dna methylation.mutants that show reduced dna methylation were identified in a mutant screen based on the assumptions that (i) the nucleoside analog 5-azacytidine (5-azac) promotes the formation of potentially lethal dna-methyltransferase adducts; (ii) reduction in dna methyltransferase will decrease the sensitivity of cells to 5-azac; and (iii) this potential selective advantage will be enhanced in mutants that are deficient in the repair of 5-azac-induced dna damage. of fifteen potential repair mutants screen ...19989738881
mapping fungal ion channel locationsion channel mapping techniques are described and the results for two fungal organisms, saprolegnia ferax and neurospora crassa, are presented. in these species, two channel types have been characterized, stretch-activated channels exhibiting significant calcium permeability and spontaneous channels having significant potassium permeability. two distinct analyses of patch clamp data, analysis of channel self-clustering and association between different channel types, and localization along the hy ...19989742193
what determines growth direction in fungal hyphae?we used high-resolution video microscopy and image analysis to map the trajectory of the spitzenkörper in growing hyphae of neurospora crassa and to correlate it with growth directionality. the spitzenkörper followed a tortuous trajectory produced by a dominant forward motion accompanied by frequent, transverse oscillations. in hyphae with a fixed growth direction, the regression line of the spitzenkörper trajectory corresponded to the longitudinal axis of the hypha. a permanent change in growth ...20079742196
agrobacterium tumefaciens-mediated transformation of filamentous fungi.agrobacterium tumefaciens transfers part of its ti plasmid, the t-dna, to plant cells during tumorigenesis. it is routinely used for the genetic modification of a wide range of plant species. we report that a. tumefaciens can also transfer its t-dna efficiently to the filamentous fungus aspergillus awamori, demonstrating dna transfer between a prokaryote and a filamentous fungus. we transformed both protoplasts and conidia with frequencies that were improved up to 600-fold as compared with conve ...19989743116
mutation of the gene for the second-largest subunit of rna polymerase i prolongs the period length of the circadian conidiation rhythm in neurospora crassa.the period length of the circadian conidiation rhythm was examined in a mutant strain of neurospora crassa, un-18, that is temperature sensitive for mycelial growth. the un-18 mutant showed a temperature-sensitive phenotype with respect to both mycelial growth and the period length of the conidiation rhythm. below 22 degrees c, the un-18 mutation did not affect the period length, but at temperatures between 22 degrees c and 32 degrees c, the period length of the un-18 mutant was approximately 2 ...19989749669
single mating type-specific genes and their 3' utrs control mating and fertility in cochliobolus heterostrophus.to determine the number of proteins required for mating type (mat) locus-regulated control of mating in cochliobolus heterostrophus, mat fragments of various sizes were expressed in mat deletion strains. as little as 1.5 kb of mat sequence, encoding a single unique protein in each mating type (mat-1 and mat-2), conferred mating ability, although an additional 160 bp of 3' utr was needed for production of ascospores. no other mating type-specific genes involved in mating identity or fertility wer ...19989749670
engineering of pyridine nucleotide specificity of nitrate reductase: mutagenesis of recombinant cytochrome b reductase fragment of neurospora crassa nadph:nitrate reductase.the cytochrome b reductase fragment of neurospora crassa nadph:nitrate reductase (ec 1.6.6.3) was overexpressed in escherichia coli with a his-tag for purification after mutation of the nadph binding site. the recombinant enzyme fragment was altered by site-directed mutagenesis guided by the three-dimensional structure of cytochrome b reductase fragment of corn nadh:nitrate reductase (ec 1.6.6.1). substitution of asp for ser920 (using residue numbering for holo-nadph:nitrate reductase of n. cras ...19989750171
a cell-free translation-translocation system reconstituted with subcellular fractions from the wall-less variant fz;sg;os-1v of neurospora crassa.a translation-translocation system reconstituted with subcellular fractions from the wall-less variant fz;sg;os-1v of neurospora crassa reproduces in vitro translocation and processing of a secretory protein. the translation extract was isolated from the wall-less variant by gently lysing cells by a freeze-thaw procedure. this method yielded more extract then the method developed previously (r. addison, j. biol. chem. 262: 17031, 1987) as well as reducing microsomal contamination. the microsomal ...19989756715
the enzymic synthesis of beta-[32p]udp-n-acetylglucosamine.cells of micrococcus sp. 2102 incorporate inorganic [32p]phosphate from the medium into the sugar-phosphate polymer of the wall. controlled acid hydrolysis of sodium dodecyl sulphate-extracted cells gives n-acetylglucosamine 6-[32p]-phosphate which can be purified by ion-exchange chromatography and incubated with utp in the presence of crude preparations of phosphoacetylglucosamine mutase from neurospora crassa and utp:n-acetylglucosamine 1-phosphate phosphotransferase from bacillus licheniformi ...19789762094
involvement of two novel chaperones in the assembly of mitochondrial nadh:ubiquinone oxidoreductase (complex i).the respiratory complex i of mitochondria consists of some 40 different subunits which form an l-shaped structure. perpendicular to a hydrophobic arm embedded in the inner mitochondrial membrane a peripheral arm protrudes into the matrix. assembly of the complex as studied in the fungus neurospora crassa involves the formation of discrete intermediates. the matrix arm and the membrane arm are formed independently of each other and are joined in the course of assembly. the membrane arm itself is ...19989769214
evidence for balancing selection operating at the het-c heterokaryon incompatibility locus in a group of filamentous fungi.in filamentous fungi, het loci (for heterokaryon incompatibility) are believed to regulate self/nonself-recognition during vegetative growth. as filamentous fungi grow, hyphal fusion occurs within an individual colony to form a network. hyphal fusion can occur also between different individuals to form a heterokaryon, in which genetically distinct nuclei occupy a common cytoplasm. however, heterokaryotic cells are viable only if the individuals involved have identical alleles at all het loci. on ...19989770498
pzl-1 encodes a novel protein phosphatase-z-like ser/thr protein phosphatase in neurospora crassa.the gene and cdna of a novel protein phosphatase were cloned from neurospora crassa. the pzl-1 gene encompasses three introns and is localized to the left arm of chromosome i between cyt-21 and fsr-12. it encodes a protein of 58.3 kda containing a ser/pro rich n-terminal segment, and a c-terminal domain that is similar to the catalytic subunit of type 1 protein phosphatases. the first 51 amino acid residues, including a potential n-myristoylation site, as well as the c-terminal domain (about 300 ...19989774742
auto-adp-ribosylation of nad glycohydrolase from neurospora crassa.nad glycohydrolase (nadase; ec 3.2.2.5) is an enzyme that catalyzes hydrolysis of nad to produce adp-ribose and nicotinamide. we recently demonstrated that self-inactivation of nadase from rabbit erythrocytes was due to an auto-adp-ribosylation. in the present study, a mechanism of self-inactivation of nadase from neurospora crassa by its substrate was investigated by using intact mycelia of n. crassa and purified nadase, which had molecular characteristics different from mammalian nadases. the ...19989787786
protein phosphatase 2a is involved in hyphal growth of neurospora crassa.cantharidin and calyculin a, natural toxins that are inhibitors of protein phosphatases 1 and 2a (ppi and pp2a, respectively). inhibit neurospora crassa hyphal growth. when n. crassa was grown in the presence of either drug, abnormalities were observed at hyphal tips. in addition, both drugs induced an increase in hyphal branching. cantharidin inhibited n. crassa hyphal growth in a temperature-dependent manner, as the effect of the drug was more pronounced at 34 degrees c than at 25 degrees c. i ...19989790584
isolation and characterization of a new gene, sre, which encodes a gata-type regulatory protein that controls iron transport in neurospora crassa.multiple gata factors - regulatory proteins with consensus zinc finger motifs that bind to dna elements containing a gata core sequence - exist in the filamentous fungus neurospora crassa. one gata factor, nit2. controls nitrogen metabolism, whereas two others, wc-1 and wc-2, regulate genes responsive to blue light induction. a gene encoding a new gata factor, named sre, was isolated from neurospora using a pcr-mediated method. sequence analysis of the new gata factor gene revealed an orf specif ...19989790585
multiple forms of arginase are differentially expressed from a single locus in neurospora crassa.the neurospora crassa catabolic enzyme, arginase (l-arginine amidinohydrolase, ec 3.5.3.1), exists in multiple forms. multiple forms of arginase are found in many vertebrates, but this is the only reported example in a microbial organism. the two major forms are structurally similar with subunit sizes of 36 and 41 kda, respectively. the larger form is produced by mycelia growing in arginine-supplemented medium. both forms are localized in the cytosol. the structural gene for arginase, aga, has b ...19989792692
identification of the first fungal annexin: analysis of annexin gene duplications and implications for eukaryotic evolution.annexin homologues have been found in animals, plants, and distinct protist lineages. we report the identification of the first fungal annexin, encoded by the anx14 gene of the filamentous ascomycete neurospora crassa. annexins have a complex evolutionary history and exhibit a large number of gene duplications and gene losses in various taxa, including the complete loss of annexin sequences from another ascomycete, the budding yeast saccharomyces cerevisiae. surprisingly, the n. crassa annexin h ...19989797403
the two-component hybrid kinase regulator canik1 of candida albicans.using degenerate primers of highly conserved regions of two-component response regulators for pcr amplification, a two-component response regulator was cloned from candida albicans that is homologous to nik-1+ of neurospora crassa. this two-component hybrid kinase, canik1, also shows features of bacterial two-component response regulators, including a putative unorthodox second histidine kinase motif at the carboxy-terminal end. canik1 was expressed at low levels in both the white and opaque swi ...19989802013
promoter analysis of the acetate-inducible isocitrate lyase gene (acu-3) from neurospora crassa.analysis of the promoter region of the acetate-induced isocitrate lyase gene (acu-3) of neurospora crassa was undertaken. a series of deletions in the 5' non-transcribed region were constructed and the effects of these mutations on the enzyme levels following growth on sucrose and transfer to acetate were measured. sequences within the region -603 to -271 relative to the transcription start site appear essential for transcription. the region -950 to -1278 is required for sucrose repression, whic ...19989804981
a new uv-sensitive mutant that suggests a second excision repair pathway in neurospora crassa.in an attempt to understand the relationship between photorepair and dark repair in neurospora crassa, a new mutant was isolated, which showed defects in both repair processes. the new mutant, mus-38, is moderately sensitive to uv and shows imperfect photoreactivation following uv irradiation. dna was purified from this mutant and the other uv-sensitive mutants, and analyzed for the removal of cyclobutane pyrimidine dimers (cpds). uv-specific endonuclease-sensitive sites (ess) completely disappe ...19989806416
carbon catabolite repression of gene expression and conidiation in neurospora crassa. 19989806802
temperature adaptation of house keeping and heat shock gene expression in neurospora crassa.adaptation of house keeping and heat shock gene expression was determined in neurospora crassa during continuous exposure to different temperatures. steady-state values of total protein synthesis differed little after incubation for 24 h at temperatures between 15 and 42 degreesc. adaptation kinetics at 42 degreesc showed an initial, transient inhibition of total protein synthesis. similar kinetics were observed with actin synthesis and tubulin mrna. a priming 1-h heat shock of 42 degreesc 2 h p ...19989806804
characterization of aspergillus nidulans peroxisomes by immunoelectron microscopy.in previous work, we have demonstrated that oleate induces a massive proliferation of microbodies (peroxisomes) in aspergillus nidulans. although at a lower level, proliferation of peroxisomes also occurs in cells growing under conditions that induce penicillin biosynthesis. here, microbodies in oleate-grown a. nidulans cells were characterized by using several antibodies that recognize peroxisomal enzymes and peroxins in a broad spectrum of eukaryotic organisms such as yeast, and plant, and mam ...19989818356
analysis of actin and actin-related protein 3 (arp3) gene expression following induction of hyphal tip formation and apolar growth in neurospora.the genes encoding actin and arp3 in the filamentous fungus neurospora crassa were cloned and sequenced. the actin structural gene is interrupted by four introns and encodes a polypeptide of 375 amino acids, which shows very high degree of identity with actin from other sources. n. crassa arp3 is 439 amino acids in length and is 71% to 80% identical to arp3s from five other organisms, while actin is 40% to 50% identical to these same arp3s. transcript levels for actin and arp3 decrease upon indu ...19989819052
the mating type locus of neurospora crassa: identification of an adjacent gene and characterization of transcripts surrounding the idiomorphs.complexity in and around the a and a mating type idiomorphs in neurospora crassa was examined. six sets of transcripts surrounding the idiomorphs were identified by northern analysis. several different patterns of regulation were observed. the two pairs of transcripts closest to the centromere-proximal idiomorph flanks (variable regions) exhibited mating type-specific size differences. dna sequence was obtained for the region surrounding the four transcripts which showed mating type-specific siz ...19989819054
the evolutionarily conserved eukaryotic arginine attenuator peptide regulates the movement of ribosomes that have translated it.translation of the upstream open reading frame (uorf) in the 5' leader segment of the neurospora crassa arg-2 mrna causes reduced initiation at a downstream start codon when arginine is plentiful. previous examination of this translational attenuation mechanism using a primer-extension inhibition (toeprint) assay in a homologous n. crassa cell-free translation system showed that arginine causes ribosomes to stall at the uorf termination codon. this stalling apparently regulates translation by pr ...19989819438
ssp1, a site-specific parvulin homolog from neurospora crassa active in protein folding.peptidyl-prolyl cis-trans-isomerases (ppiases) are enzymes capable of isomerizing a xaa-pro peptide bond. three families of ppiases are known: cyclophilins, fkbps, and parvulins. the physiological functions of the ppiases are only poorly understood. eucaryotic members of the parvulin family have recently been shown to be essential for regulation of mitosis. here we describe the purification and characterization of ssp1, an abundant parvulin homolog from neurospora crassa, which is unique among t ...19989822668
isolation and characterization of alpha-tubulin genes from septoria tritici and rhynchosporium secalis, and comparative analysis of fungal alpha-tubulin sequences.the alpha-tubulin genes from septoria tritici and rhynchosporium secalis have been cloned and sequenced. the predicted amino acid sequence and intron structure showed strong homology with other known filamentous fungal alpha-tubulins. comparison of sixteen fungal alpha-tubulin sequences based on amino acid sequence homology and intron structure identified five groups of proteins. group 1 consists of filamentous fungi, including s. tritici and r. secalis, the dimorphic fungus histoplasma capsulat ...19989829778
characterization of a gene encoding a single-subunit bacteriophage-type rna polymerase from maize which is alternatively spliced.single-subunit rna polymerases belonging to the t3/t7 bacteriophage family are thought to be common throughout eukaryotes. we report the isolation and characterization of a nucleus-encoded single-subunit rna polymerase gene from maize. this gene is highly homologous to other single-subunit rna polymerase genes from arabidopsis, chenopodium. yeast and neurospora crassa involved in organellar transcription. genomic southern analysis reveals 10 to 15 hybridising fragments, suggesting that maize con ...19989829825
differential stress gene expression during the development of neurospora crassa and other fungi.stress genes are differentially expressed during the development of neurospora crassa and other fungi. large amounts of constitutive heat shock protein 70 (hsc70) are found in dormant conidia of n. crassa, whereas little mrna of the related glucose-regulated protein (grp78) is detected. it is, however, not generally clear whether heat shock protein or mrna is preferentially stored in dormant spores. germinating spores of n. crassa increase the level of grp mrna. during this developmental stage, ...19989835024
mitop: database for mitochondria-related proteins, genes and diseases.the mitop database http://websvr.mips.biochem.mpg. de/proj/medgen/mitop/ consolidates information on both nuclear- and mitochondrial-encoded genes and their proteins. the five species files- saccharomyces cerevisiae, mus musculus, caenorhabditis elegans, neurospora crassa and homo sapiens -include annotated data derived from a variety of online resources and the literature. a wide spectrum of search facilities is given in the interelated sections 'gene catalogues', 'protein catalogues', 'homolog ...19999847163
spermidine determines the sensitivity to the calmodulin antagonist, chlorpromazine, for the circadian conidiation rhythm but not for the mycelial growth in neurospora crassa.the gene that suppresses the phenotype of the cpz-2 mutation, which results in changing the sensitivity to chlorpromazine in relation to mycelial growth and circadian rhythms, was cloned in neurospora crassa. this gene is not the cpz-2 gene itself but rather is identical to the spe-3 gene that encodes spermidine synthase in neurospora. the intracellular content of spermidine was lowered in the cpz-2 strain compared to that of the wild-type strain. by integration of the spe-3 gene or by the addit ...19989850006
isolation of a class iv chitin synthase gene from a zygomycete fungus, rhizopus oligosporus.we found the presence of dna sequence which shows sequence similarity to the class iv chitin synthase gene (chs3) of saccharomyces cerevisiae in the genome of 14 rhizopus species which belong to zygomycetes. we cloned a gene (chs3), which might correspond to one of these homologous sequences, from rhizopus oligosporus by low stringency plaque hybridization probed with chs3. the deduced amino acid sequence of this gene showed highest similarity to the class iv chitin synthase of neurospora crassa ...19989851030
isolation of a nitrogen response regulator gene (nrr1) from metarhizium anisopliae.attempts to improve the effectiveness of entomopathogenic fungi as biological control agents require a clear understanding of the pathogenicity determinants at both the biochemical and molecular level. proteases play a key role in entomopathogenicity, allowing the fungus to penetrate the insect cuticle and rapidly invade the host. the most extensively studied of these protease activities, pr1a and pr2, are both subject to nitrogen derepression. the metarhizium anisopliae nrr1 (nitrogen response ...19989852945
ancf, the ccaat binding complex of aspergillus nidulans, contains products of the hapb, hapc, and hape genes and is required for activation by the pathway-specific regulatory gene amdr.ccaat binding factors (cbfs) positively regulating the expression of the amds gene (encoding acetamidase) and two penicillin biosynthesis genes (ipna and aata) have been previously found in aspergillus nidulans. the factors were called ancf and penr1, respectively. deletion of the hapc gene, encoding a protein with significant similarity to hap3p of saccharomyces cerevisiae, eliminated both ancf and penr1 binding activities. we now report the isolation of the genes hapb and hape, which encode pr ...19999858535
phosphate transporter gene family of arabidopsis thaliana.using a high-affinity phosphate transporter gene of arabidopsis thaliana, pht1, as a probe, three arabidopsis homologs were isolated, named pht4, pht5 and pht6, in addition to the previously isolated pht2 and pht3. the amino acid sequences deduced from the three nucleotides were 32-42% homologous with microbial phosphate transporters of saccharomyces cerevisiae (pho84), neurospora crassa (pho-5) and glomus versiforme (gvpt). pht1, pht2, pht3 and pht6 were clustered in a 25-kbp region of chromoso ...19989872450
mutational activation of a galphai causes uncontrolled proliferation of aerial hyphae and increased sensitivity to heat and oxidative stress in neurospora crassa.heterotrimeric g proteins, consisting of alpha, beta, and gamma subunits, transduce environmental signals through coupling to plasma membrane-localized receptors. we previously reported that the filamentous fungus neurospora crassa possesses a galpha protein, gna-1, that is a member of the galphai superfamily. deletion of gna-1 leads to defects in apical extension, differentiation of asexual spores, sensitivity to hyperosmotic media, and female fertility. in addition, deltagna-1 strains have low ...19999872952
molybdenum-free nitrate reductases from vanadate-reducing bacteria.two catalytically distinct molybdenum-free dissimilatory nitrate reductases, a soluble periplasmic and a membrane-bound one, were isolated from the vanadate-reducing facultatively anaerobic bacterium pseudomonas isachenkovii and purified to electrophoretic homogeneity. the enzymes did not contain molybdenum, the periplasmic enzyme contained vanadium, whereas the membrane-bound enzyme was vanadium-free. both nitrate reductases lacked molybdenum cofactor. this fact was proved by reconstitution of ...19989883895
radioactivity-based and spectrophotometric assays for isoorotate decarboxylase: identification of the thymidine salvage pathway in lower eukaryotes.a few organisms, notably some fungi, have the ability to metabolize thymidine to uracil, thus conserving the pyrimidine ring for subsequent metabolic use. neurospora crassa possesses this pathway, termed the thymidine salvage pathway, and can utilize thymidine as a total pyrimidine source. the enzyme isoorotate decarboxylase (idcase) completes this pathway via the enzymatic removal of the carboxylate from isoorotate to yield uracil. we describe in this communication two assays for idcase and the ...19999887216
folylpolyglutamate synthesis in neurospora crassa: primary structure of the folylpolyglutamate synthetase gene and elucidation of the met-6 mutation.in neurospora crassa, the met-6+ gene encodes folylpoly-gamma-glutamate synthetase (fpgs) which catalyzes the formation of polyglutamate forms of folate. methionine auxotrophy of the neurospora crassa met-6 mutant is related to a lesion affecting this enzyme. functional complementation of the mutant strain was achieved by introducing copies of the wild-type met-6+ gene into mutant spheroplasts. the complementing sequences were found to be contained on a 3.5 kb ecori-bamhi restriction fragment. t ...19989887523
search for novel redox groups in mitochondrial nadh:ubiquinone oxidoreductase (complex i) by diode array uv/vis spectroscopy.the proton-translocating nadh:ubiquinone oxidoreductase of mitochondria (complex i) is a large l-shaped multisubunit complex. the peripheral matrix arm contains one fmn and a number of iron-sulfur (fes) clusters and is involved in nadh oxidation and electron transfer to the membrane intrinsic arm. there, following a yet unknown mechanism, the redox-driven proton translocation and the ubiquinone reduction take place. redox groups that would be able to link electron transfer with proton translocat ...19989914816
chitin synthase genes in the arbuscular mycorrhizal fungus glomus versiforme: full sequence of a gene encoding a class iv chitin synthase.chitin synthase genes of the arbuscular mycorrhizal fungus glomus versiforme were sought in an investigation of the molecular basis of fungal growth. three dna fragments (gvchs1, gvchs2 and gvchs3) corresponding to the conserved regions of distinct chitin synthase (chs) genes were amplified by means of the polymerase chain reaction (pcr) with two sets of degenerate primers. gvchs1 and gvchs2 encode two class i chitin synthases, whereas gvchs3 encodes a class iv chitin synthase. a genomic library ...19999919652
molecular characterization of tol, a mediator of mating-type-associated vegetative incompatibility in neurospora crassa.the mating-type locus in the haploid filamentous fungus, neurospora crassa, controls mating and sexual development. the fusion of reproductive structures of opposite mating type, a and a, is required to initiate sexual reproduction. however, the fusion of hyphae of opposite mating type during vegetative growth results in growth inhibition and cell death, a process that is mediated by the tol locus. mutations in tol are recessive and suppress mating-type-associated heterokaryon incompatibility. i ...19999927450
cloning and expression of the cdna encoding an alternative oxidase gene from aspergillus niger wu-2223l.a cdna fragment encoding the mitochondrial alternative oxidase, the enzyme responsible for cyanide-insensitive and salicylhydroxamic acid (sham)-sensitive respiration, from the citric acid-producing fungus aspergillus niger wu-2223l was cloned and expressed in escherichia coli as a host strain. synthetic primers were designed from the conserved nucleotide sequences of the alternative oxidase genes from higher plants and a yeast. the 210-bp dna fragment was amplified by pcr with these primers usi ...19999933359
the b regulatory subunit of protein phosphatase 2a is required for completion of macroconidiation and other developmental processes in neurospora crassa.rgb-1, encoding the tentative b regulatory subunit of the type 2a ser/thr phosphatase in neurospora crassa, was isolated from cdna and genomic libraries. based on analysis of cdna and genomic clones, rgb-1 is 3387 nucleotides in length, contains seven putative introns and encodes a 461-amino-acid polypeptide. intron i, which is 5' to the presumed translation initiation codon, contains a uorf encoding 34 amino acids. intron vi undergoes alternative splicing. inactivation of rgb-1 by the repeat-in ...19999987122
the aspergillus nidulans gata factor srea is involved in regulation of siderophore biosynthesis and control of iron uptake.a gene encoding a new gata factor from aspergillus nidulans, srea, was isolated and characterized. srea displays homology to two fungal regulators of siderophore biosynthesis: about 30% overall identity to sre from neurospora crassa and about 50% identity to urbs1 from ustilago maydis over a stretch of 200 amino acid residues containing two gata-type zinc finger motifs and a cysteine-rich region. this putative dna binding domain, expressed as a fusion protein in escherichia coli, specifically bi ...19999988696
cloning of two genes encoding potassium transporters in neurospora crassa and expression of the corresponding cdnas in saccharomyces cerevisiae.two neurospora crassa genes, trk-1 and hak-1, encode k+ transporters that show sequence similarities to the trk transporters described in saccharomyces cerevisiae and schizosaccharomyces pombe, and to the hak transporters described in schwanniomyces occidentalis and barley. the n. crassa trk1 and hak1 transporters expressed by the corresponding cdnas in a trk1 delta trk2 delta mutant of s. cerevisiae exhibited a high affinity for rb+ and k+. northern blot analysis and comparison of the kinetic c ...199910027968
the goodwin model: simulating the effect of cycloheximide and heat shock on the sporulation rhythm of neurospora crassa.the goodwin model is a negative feedback oscillator which describes rather closely the putative molecular mechanism of the circadian clock of neurospora and drosophila. an essential feature is that one or two clock proteins are synthesized and degraded in a rhythmic fashion. when protein synthesis in n. crassa (wild-type frq+and long-period mutant frq7) was inhibited by continuous incubation with increasing concentrations of cycloheximide (chx) the period of the circadian sporulation rhythmicity ...199910036201
the wd protein cpc2p is required for repression of gcn4 protein activity in yeast in the absence of amino-acid starvation.the cpc2 gene of the budding yeast saccharomyces cerevisiae encodes a g beta-like wd protein which is involved in regulating the activity of the general control activator gcn4p. the cpc2 gene encodes a premrna which is spliced and constitutively expressed in the presence or absence of amino acids. loss of cpc2 gene function suppresses a deletion of the gcn2 gene encoding the general control sensor kinase, but not a deletion in the gcn4 gene. the resulting phenotype has resistance against amino-a ...199910048025
isolation, characterization and disruption of the area nitrogen regulatory gene of gibberella fujikuroi.the gene area-gf, a homologue of the major nitrogen regulatory genes nit-2, area, nre and nut1 of neurospora crassa, aspergillus nidulans, penicillium chrysogenum and magnaporthe grisea, respectively, was cloned from the gibberellin (ga)-producing rice pathogen gibberella fujikuroi. area-gf encodes a protein of 972 amino acid residues which contains a single putative zinc finger dna-binding domain that is at least 98% identical to the zinc finger domains of the homologous fungal proteins. the ar ...199910071216
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