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genetic regulation of the quinic acid utilization (qut) gene cluster in aspergillus nidulans.a large number of quinic acid non-utilizing qut mutants of aspergillus nidulans deficient in the induction of all three quinic acid specific enzymes have been analysed. one class the qutd mutants, are all recessive and are non-inducible at ph 6.5 due to inferred deficiency in a quinate ion permease. two regulatory genes have been identified. the quta gene encodes an activator protein since most quta mutants are recessive and non-inducible although a few fully dominant mutants have been found. th ...19883049934
the yeast aminopeptidase y.a metal-dependent aminopeptidase (ec 3.4.11.-), designated apase y, has been purified to homogeneity by conventional methods. the enzyme is composed of a single polypeptide chain with molecular mass of 102 kilodaltons, estimated by sodium dodecyl sulphate - polyacrylamide gel electrophoresis, with a blocked n-terminal amino acid. it possesses neither endopeptidase nor carboxypeptidase activity and is strongly inhibited by metal-chelating agents, zn2+, and the protein inhibitor from neurospora cr ...19883042113
regulation of fructose 2,6-bisphosphate levels in neurospora crassa.both wild type and cr-1 mutant (adenylate cyclase and cyclic amp-deficient) strains of neurospora crassa contain fructose 2,6-bisphosphate at levels of 27 nmol/g dry tissue weight. this level decreases by about 50% in both strains upon depriving the cells of carbon or nitrogen sources for 3 h. an increase in cyclic amp levels produced by addition of lysine to nitrogen-starved cells produced no increase in fructose 2,6-bisphosphate levels. both strains respond to short-term addition of salicylate ...20083040112
monophenol monooxygenase from neurospora crassa. 19873037257
isolation and characterization of the nuclear gene encoding the rieske iron-sulfur protein (rip1) from saccharomyces cerevisiae.the nuclear gene encoding the rieske iron-sulfur protein of the cytochrome bc1 complex of the mitochondrial respiratory chain has been isolated and characterized from saccharomyces cerevisiae. we used a segment of the iron-sulfur protein gene from neurospora crassa (harnisch, u., weiss, h., and sebald, w. (1985) eur. j. biochem. 149, 95-99) to detect the yeast gene by southern analysis. five different but overlapping clones were then isolated by probing a yeast genomic library carried on yep 13 ...19873036836
some aspects of the regulation of pyruvate kinase levels in neurospora crassa.pyruvate kinase levels were monitored in neurospora crassa mycelium (grown on different carbon sources for varying time intervals) by immunoprecipitation using polyclonal antibodies raised against a purified enzyme preparation. pyruvate kinase specific mrna was demonstrated by hybridization of northern and dot blots of total rna with a n. crassa pyruvate kinase gene fragment. two pyruvate kinase specific mrna species were detected in mycelia of all ages examined. an age-dependent and carbon sour ...19873036325
structure and expression of the overlapping nd4l and nd5 genes of neurospora crassa mitochondria.genes homologous to the mammalian mitochondrial nadh dehydrogenase subunit genes nd4l and nd5 were identified in the mitochondrial genome of the filamentous fungus neurospora crassa, and the structure and expression of these genes was examined. the nd4l gene (interrupted by one intervening sequence) potentially encodes an 89 residue long hydrophobic protein that shares about 26% homology (or 41% homology if conservative amino acid substitutions are allowed) with the analogous human mitochondrial ...19873035337
a single precursor protein for two separable mitochondrial enzymes in neurospora crassa.the arg-6 locus of neurospora crassa encodes two early enzymes of the arginine biosynthetic pathway, acetylglutamate kinase and acetylglutamyl-phosphate reductase. previous genetic and biochemical analyses of this locus and its products showed that: 1) strains carrying polar nonsense mutations in the acetylglutamate kinase gene lacked both enzyme activities (davis, r.h., and weiss, r.l. (1983) mol. gen. genet. 192, 46-50), and 2) the proteins isolated from mitochondria were completely separable ...19873032945
gmp-stimulation of the cyanide-insensitive mitochondrial respiration in heat-shocked conidia of neurospora crassa.in mitochondria of heat-shocked conidia of neurospora exogenous nadh and succinate were oxidized mainly via the alternative, hydroxamate-sensitive pathway (70%) and only 30% via the cytochromic, cyanide-sensitive pathway which was predominant in untreated conidia; the alternative oxidase pathway was markedly stimulated by guanosine 5'-monophosphate (gmp).19873032673
characterization of two abundantly expressed constitutive genes of neurospora crassa.two abundantly expressed, constitutive genes of neurospora crassa were isolated during differential screening of neurospora genomic libraries. the coding regions of these two genes, designated rlf1 and rlf3, were identified by hybridization of the cloned dna sequences with cdna probes made from polyadenylated rna. the rlf3 gene was carried on a 15-kilobase neurospora bamhi dna fragment present in a lambda 1059 recombinant; a 2-kilobase restriction fragment that contains rlf3 was subcloned into p ...19873032385
molecular cloning of a cdna for a human adp/atp carrier which is growth-regulated.we have identified in a human cdna library a clone (hp2f1) whose cognate rna is growth-regulated. the insert has been sequenced and the nucleotide sequence shows a strong homology to the nucleotide sequences of the adp/atp carrier cdna and gene, respectively, isolated from neurospora crassa and saccharomyces cerevisiae. the putative amino acid sequence of hp2f1 shows an 87% homology to the amino acid sequence of the adp/atp carrier from beef heart mitochondria. we conclude that the insert of hp2 ...19873031073
import of cytochrome c into mitochondria. cytochrome c heme lyase.the import of cytochrome c into mitochondria can be resolved into a number of discrete steps. here we report on the covalent attachment of heme to apocytochrome c by the enzyme cytochrome c heme lyase in mitochondria from neurospora crassa. a new method was developed to measure directly the linkage of heme to apocytochrome c. this method is independent of conformational changes in the protein accompanying heme attachment. tryptic peptides of [35s]cysteine-labelled apocytochrome c, and of enzymat ...19873030750
the primary structure of cytochrome c1 from neurospora crassa.the primary structure of the cytochrome c1 subunit of ubiquinol-cytochrome-c reductase from mitochondria of neurospora crassa was determined by sequencing the cdna of a bank cloned in escherichia coli. from the coding region the sequence of 332 amino acids, corresponding to the molecular mass of 36,496 da, was derived for the precursor protein. the mature protein, the n terminus of which was previously sequenced [tsugita et al. (1979) in cytochrome oxidase (king, t. e. et al., eds) pp. 67-77, el ...19873030747
purification and properties of a single strand-specific endonuclease from mouse cell mitochondria.a nuclease was purified from mitochondria of the mouse plasmacytoma cell line, mcp-11 which acts on single-stranded dna endonucleolytically and appears to have no activity upon native dna. it degrades unordered rna somewhat more effectively than it does dna. the enzyme activity and the major detectable polypeptide migrate to a position corresponding to an mr of 37,400 on denaturing polyacrylamide gels; in its native form the activity has an s value of 4.7, which corresponds to a molecular weight ...19863027656
alkaline protease deficiency in the cr-1 (crisp) mutant of neurospora crassa.the adenyl cyclase deficient cr-1 mutant of neurospora crassa grew poorly in bovine serum albumin as an alternative and only source of either sulfur, nitrogen or carbon. the low growth of the cr-1 mutant in protein was correlated with limited secretion of extracellular alkaline protease. the defect was specific for the cr-1 mutant and was suppressed by exogenous cyclic amp. cyclic amp relieved protease deficiency under carbon, nitrogen or sulfur limiting conditions to unequal extents. protease s ...19863026533
an upstream signal is required for in vitro transcription of neurospora 5s rna genes.the dna sequences upstream of the 5s rna genes in neurospora crassa are largely different from one another, but share a short consensus sequence located in the segment 29 to 26 nucleotides preceding the transcribed region. differences among flanking sequences do not appear to affect transcription. deletion analysis indicates, however, that a dna segment including the conserved "tata box" is required for in vitro transcription of neurospora 5s rna genes.19863025558
nuclear endo-exonuclease of neurospora crassa. evidence for a role in dna repair.the major nuclease activity in nuclei of mycelia of neurospora crassa has been identified as that of endoexonuclease, an enzyme purified and characterized previously from mitochondria and vacuoles which acts endonucleolytically on single-stranded dna and rna and possesses highly processive exonuclease activity with double-stranded dna. cross-contamination from the other organelles was eliminated as a source of the activity. endo-exonuclease of nucleoplasm, chromatin, and nuclear matrix showed 80 ...19873025215
neurospora crassa ribosomal dna: sequence of internal transcribed spacer and comparison with n. intermedia and n. sitophila.using [32p]dna probes from a clone containing 17s, 5.8s and 26s rrna of neurospora crassa, the remainder of the repeat unit (ru) for ribosomal dna (rdna) has been cloned. combining restriction analysis of the cloned dna and restriction digests of genomic dna, the ru was found to be 8.7 kb. the nucleotide sequence was determined for the internal transcribed spacer (its) regions one and two, for 5.8s rrna and for portions of 17s and 26s rrnas immediately flanking the its regions, and compared to t ...19863021585
pyrophosphate-caused inhibition of the aminoacylation of trna by the leucyl-trna synthetase from neurospora crassa.inorganic pyrophosphate inhibits the aminoacylation of trnaleu by the leucyl-trna synthetase from neurospora crassa giving very low kapp.i, ppi values of 3-20 microm. the inhibition by pyrophosphate, together with earlier kinetic data, suggest a reaction mechanism where leucine, atp and trna are bound to the enzyme in almost random order, and pyrophosphate is dissociated before the rate-limiting step. a kinetic analysis of this mechanism shows that the measured kapp.i values do not give the real ...19863021454
sequence analysis of the pyr-4 (orotidine 5'-p decarboxylase) gene of neurospora crassa.the pyr-4 gene of neurospora crassa encodes orotidine-5' -phosphate decarboxylase, which catalyses the sixth step in the pyrimidine biosynthetic pathway. the complete nucleotide sequence of a 1.8-kb genomic fragment containing the pyr-4 gene has been determined. using transposon mutagenesis, the coding region has been identified, and the amino acid (aa) sequence deduced. comparison of the pyr-4 aa sequence with ura3, the equivalent gene of saccharomyces cerevisiae, showed extensive blocks of hom ...19863019837
genetical and molecular analyses of qa-2 transformants in neurospora crassa.neurospora crassa qa-2+ transformants from five different donor dna clones were analyzed by genetical and molecular techniques. none of the 32 transformants have the qa-2+ dna replacing the qa-2- gene in linkage group vii. in one transformant, the qa-2+ dna was inserted adjacent to the qa-2- gene. thirty-one transformants have the qa-2+ inserts at sites not linked, or not closely linked, to the qa-2 locus in lg vii. plasmid sequences were integrated along with the qa-2+ gene in 28 transformants. ...19863015719
[nad+-kinase from neurospora crassa: isolation and properties].the nad+ kinase (ec 2.7.1.23) of the filamentous fungus n. crassa is localized in cytosol. the activity in the dialyzed cell free extract has a ph optimum 8.3; it utilizes only atp but not inorganic polyphosphates as a phosphoryl donor. a method for 200-fold purification of nad+ kinase with a 20% yield has been developed. the procedure includes 105000 g centrifugation, fractionation with (nh4)2so4, isoelectrofocusing in a ultrodex layer and preparative electrophoresis in polyacrylamide gel. the ...19863015266
a potassium-proton symport in neurospora crassa.combined ion flux and electrophysiological measurements have been used to characterized active transport of potassium by cells of neurospora crassa that have been moderately starved of k+ and then maintained in the presence of millimolar free calcium ions. these conditions elicit a high-affinity (k1/2 = 1-10 microm) potassium uptake system that is strongly depolarizing. current-voltage measurements have demonstrated a k+-associated inward current exceeding (at saturation) half the total current ...20113014042
an immunochemical study of neurospora nucleases.nucleases derived from neurospora crassa mycelia with neutral single-strand (ss) endodeoxyribonuclease activity have been examined by immunochemical techniques and by sodium dodecyl sulfate - dna gel electrophoresis. all of the intracellular nucleases, which have different divalent metal ion requirements, different strand specificities with single- and double-strand dna, different modes of action on dna and rna, and other distinguishing characteristics, are immunochemically related to neurospora ...19863013242
cloning of the arg-12 gene of neurospora crassa and regulation of its transcript via cross-pathway amino acid control.the arg-12 locus of neurospora crassa encodes ornithine carbamoyl transferase, which is one of many amino acid synthetic enzymes whose activity is regulated through cross-pathway (or general) amino acid control. we report here the use of probes derived from the functionally equivalent arg-b gene of aspergillus nidulans to identify and clone a 10 kb neurospora dna fragment carrying the arg-12 gene. short neurospora dna probes derived from this fragment were used to identify a 1.5 kb polya+ transc ...19863012277
the e35 stopper mutant of neurospora crassa: precise localization of deletion endpoints in mitochondrial dna and evidence that the deleted dna codes for a subunit of nadh dehydrogenase.two types of defective mitochondrial dna molecules with large deletions (5 kbp and 40 kbp) have previously been identified in the stopper mutant, e35, of neurospora crassa. the junction fragments spanning the deletion endpoints have now been cloned and sequenced, and their sequences compared with those of the corresponding wild-type fragments. we show that both types of defective mitochondrial dnas result from deletions of sequences flanked by short direct repeats, which are themselves parts of ...19863011426
primary structure and spectroscopic studies of neurospora copper metallothionein.when neurospora crassa is grown in the presence of cu(ii) ions, it accumulates the metal with the concomitant synthesis of a low molecular weight copper-binding protein. the molecule binds 6 g-atom of copper per mole protein (mr = 2200) and shows a striking sequence homology to the zinc- and cadmium-binding vertebrate metallothioneins. absorption, circular dichroism, and electron paramagnetic resonance spectroscopy of neurospora metallothionein indicate the copper to be bound to cysteinyl residu ...19863011391
the qa repressor gene of neurospora crassa: wild-type and mutant nucleotide sequences.the qa-1s gene, one of two regulatory genes in the qa gene cluster of neurospora crassa, encodes the qa repressor. the qa-1s gene together with the qa-1f gene, which encodes the qa activator protein, control the expression of all seven qa genes, including those encoding the inducible enzymes responsible for the utilization of quinic acid as a carbon source. the nucleotide sequence of the qa-1s gene and its flanking regions has been determined. the deduced coding sequence for the qa-1s protein en ...20103010294
elevation of alu i-induced frequencies of chromosomal aberrations in chinese hamster ovary cells by neurospora crassa endonuclease and by ammonium sulfate.the frequencies of chromosomal aberrations induced by the restriction endonuclease alu i (recognition site ag/ct) can be elevated to a similar extent by additional treatments with a single-strand-specific endonuclease from neurospora crassa (ec 3.1.30.1), or with ammonium sulfate in which the neurospora endonuclease is suspended. these data indicate that alu i does not produce dna single-strand breaks in the chromatin of living cells, which can be recognized by the neurospora endonuclease. the s ...19863010098
gene cloning in aspergillus nidulans: isolation of the isocitrate lyase gene (acud).an aspergillus nidulans gene library was constructed in a high-frequency transformation vector, pdjb3, based on the neurospora crassa pyr4 gene. this gene library was used to isolate the structural gene for isocitrate lyase (acud) by complementation of a deficiency mutation following transformation of a. nidulans. plasmids rescued in escherichia coli were able to transform five different a. nidulans acud mutants. transformation using plasmids containing the cloned fragment resulted in integratio ...19863010050
a missense mutation in the oxi-3 gene of the [mi-3] extranuclear mutant of neurospora crassa.we have determined the dna sequence of the oxi-3 gene and its 5' flanking region in the extranuclear [mi-3] mutant of neurospora crassa. the oxi-3 gene encodes subunit 1 of cytochrome c oxidase, a protein known to be altered in the [mi-3] mutant (bertrand, h., and werner, s. (1979) eur. j. biochem. 98, 9-18). when the sequence from [mi-3] was compared to previously published sequences of the same region of mtdna from wild-type n. crassa, a total of five differences was found. four of these diffe ...19863007516
cloning and characterization of the multifunctional his-3 gene of neurospora crassa.we have cloned the his-3 gene of neurospora crassa and determined its nucleotide sequence. the gene specifies a protein of 863 amino acids (aa) and contains a 59-bp intron which interrupts aa 800, a proline residue. the 5' end of the his-3 transcript is heterogeneous with major starts 122 and 124 bp upstream from the start codon. there are three possible polyadenylation sites, 119, 120 and 121 bp after the uaa stop codon. the protein shows two regions of homology to the yeast his4 gene which cor ...19853005109
characterization of neurospora crassa cyclic amp phosphodiesterase activated by calmodulin.activation of cyclic amp phosphodiesterase i by brain or neurospora calmodulin was studied. the stimulation required micromolar concentrations of ca2+, and it was observed at cyclic amp concentrations between 0.1 and 500 microm. activation was blocked by edta and some neuroleptic drugs such as chlorpromazine and fluphenazine. these drugs inhibit the elongation of n. crassa wild-type aerial hyphae. these results reinforce the evidence towards the recognition of ca2+-calmodulin as one of the syste ...19853004404
the mitochondrial urf1 gene in neurospora crassa has an intron that contains a novel type of urf.in neurospora crassa, a 2670 base-pair segment of the mitochondrial dna was sequenced including a gene homologous to the mammalian urf1 that was recently shown to encode a subunit of the respiratory chain nadh dehydrogenase complex. urf1 of n. crassa is interrupted by an intron of 1118 base-pairs that divides the protein-coding sequence into two exons of 636 and 480 base-pairs length, respectively. the deduced urf1 polypeptide of 371 residues was aligned with that of other eukaryotes, revealing ...19853003362
primary structure of tyrosinase from streptomyces glaucescens.the complete amino acid sequence of streptomyces glaucescens tyrosinase is reported. the molecule consists of 273 amino acids and has a mr of 30 900 including two copper atoms. the primary structure was determined by a combination of amino acid and dna sequence analysis. peptide sequence information was derived from the cyanogen bromide, tryptic, and thermolytic fragments of apotyrosinase by automated edman degradation and aminopeptidase m and carboxypeptidase c digestions. the nucleotide sequen ...19853002431
nuclear genes for cytochrome c oxidase subunits of neurospora crassa. isolation and characterization of cdna clones for subunits iv, v, vi, and possibly vii.we obtained cdna clones for cytochrome oxidase subunits iv, v, vi, and possibly vii by constructing a lambda gt11 library of neurospora crassa cdna and probing it with antiserum directed against neurospora cytochrome oxidase holoenzyme. positive clones were further characterized with antisera directed against individual cytochrome oxidase subunits and subsequently by dna sequencing. the clones for subunits iv and v encode proteins with regions matching the known n-terminal amino acid sequences o ...19863001085
development of a high-frequency transforming vector for aspergillus nidulans.the pyr4 gene of neurospora crassa, which codes for orotidine-5'-phosphate decarboxylase, is capable of transforming an aspergillus nidulans pyrg mutant by chromosomal integration, despite low homology between the transforming dna and the recipient genome. integration of pfb6, a plasmid carrying pyr4 and capable of replication in escherichia coli, was not observed at the pyrg locus. the efficiency of transformation was considerably enhanced (50-100 fold) by inclusion in the transforming vector o ...19853000883
correct removal by splicing of a neurospora intron in yeast.processing of intron-containing nuclear messenger rnas in yeast require an internal conserved sequence (ics) element, uacuaac. similar elements (ugcuagac) have been identified in sequences interrupting nuclear genes of the related ascomycete neurospora crassa. to examine the structural splicing requirements in yeast, we constructed hybrid genes containing the intron of the neurospora histone h3 gene and cloned them into high copy number yeast vectors. subsequently we analyzed the rnas transcribe ...19852999703
calmodulin and ca2+-dependent cyclic amp phosphodiesterase activity in trypanosoma cruzi.calmodulin has been purified from trypanosoma cruzi epimastigote forms by ion-exchange chromatography, gel filtration and affinity chromatography on 2-chloro-10-(3-aminopropyl)phenotiazine-sepharose. upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the factor showed a polypeptide band with an apparent molecular weight of 16 000. in addition, cyclic amp phosphodiesterase activity from t. cruzi epimastigote forms was purified by ion-exchange chromatography and affinity chromatograph ...19852999589
excision-amplification of mitochondrial dna during senescence in podospora anserina. dna sequence analysis of three unique "plasmids".during senescence in the filamentous fungus podospora anserina, specific regions of the mitochondrial genome, termed sendna are excised, ligated and amplified. we have cloned in their entirety three such autonomously replicating plasmids, alpha, beta and epsilon sendna. none of these plasmids displayed cross-hybridization nor did we detect any significant dna homology by computer analysis. the complete dna sequence of the 2.5 kb alpha, the 5.5 kb epsilon and about 3.4 kb of the 9.8 kb beta sendn ...19852997455
neurospora crassa and s1 nuclease cleavage in hsp 83 gene chromatin.we have examined the distribution of neurospora crassa and s1 nuclease cleavage products in the chromatin of the hsp 83 heat shock gene from the drosophila melanogaster cytogenetic locus 63 bc. both of these nucleases generate double strand breaks in chromatin at specific sites close to the 5' end of the hsp 83 gene. with n. crassa nuclease we observe one major 5' fragment which is derived from nuclease cleavage in a dna segment mapping approximately 120 base-pairs from the beginning of the tran ...19852995684
isolation and sequence analysis of a cdna encoding the atp/adp translocator of zea mays l.a cdna complementary to the mrna for the atp/adp translocator of maize (zea mays l.) has been identified by virtue of hybridisation with the homologous gene from yeast. the cloned cdna has been shown by dna sequence analysis to contain an open reading frame of 954bp., which encodes a polypeptide of molecular weight 40,519. this polypeptide exhibits a high degree of homology to the translocator polypeptides of beef heart and neurospora crassa mitochondria.19852994015
cloning of the aro cluster gene of neurospora crassa and its expression in escherichia coli.we have constructed a phage, lambda ncl, which comprises a 4.0 kb hindiii insert of neurospora dna into the immunity region of the vector lambda 598. lambda ncl complements the arod6 mutation of e. coli, permitting the formation of galaxy plaques on medium lacking aromatic supplements, and transforms an aro-9 qa-2 neurospora mutant to prototrophy at a low frequency. low levels of 5-dehydroquinate hydrolyase (e.c.4.2.1.10.), with properties unlike those of the catabolic isoenzyme that is coded by ...19852993794
comparison of the mitochondrial endonucleases from neurospora crassa and saccharomyces cerevisiae.the endonucleases from neurospora crassa and saccharomyces cerevisiae are not closely related antigenically. they also differ with respect to their activity at ph 8, their degree of hydrophobicity, and their sensitivity to elevated temperatures. however, the two nucleases have similar specific activities, are inhibited by edta, and have nearly identical substrate specificities. since the enzymes also have the same mode of action and intracellular location, these similarities may indicate that th ...19852992732
a regulatory protein for orthophosphate-regulated cyclic phosphodiesterase in neurospora crassa.a regulatory protein for orthophosphate-regulated cyclic phosphodiesterase (cpdase) was detected in mycelial extracts of neurospora crassa. the protein, designated neucrassin, was precipitated by ammonium sulfate between 60 to 100% saturation, and fractionated by gel filtration through a tsk-gel column. the molecular weight was estimated to be 65,000. neucrassin inhibited the hydrolyzing activity of cpdase for cyclic 3',5'-amp in the presence of mncl2 in a noncompetitive manner, whereas it stimu ...19852992492
accurate transcription of cloned neurospora rna polymerase ii-dependent genes in vitro by homologous soluble extracts.we have developed soluble extracts from neurospora crassa capable of accurately initiating the transcription of cloned neurospora protein-encoding genes by rna polymerase ii in vitro. the genes encoding glutamate dehydrogenase (am) and histones h3 and h4 were transcribed by the extracts, and transcription was sensitive to alpha-amanitin at 1 mg/ml. the 5' heterogeneity of the in vitro initiation reactions was highly specific. of the 17 transcription initiation sites within the inducible qa gene ...19852991927
cloning and expression of the fbc operon encoding the fes protein, cytochrome b and cytochrome c1 from the rhodopseudomonas sphaeroides b/c1 complex.the gene for the fes protein of the rhodopseudomonas sphaeroides b/c1 complex was identified by means of cross-hybridization with a segment of the gene encoding the corresponding fes protein of neurospora crassa. plasmids (prsf1-14) containing the cross-hybridizing region, covering in total 13.5 kb of chromosomal dna, were expressed in vitro in a homologous system. one rsf plasmid directed the synthesis of all three main polypeptides of the r. sphaeroides b/c1 complex: the fes protein, cytochrom ...19852990901
rna processing in neurospora crassa mitochondria: use of transfer rna sequences as signals.we have used rna gel transfer hybridization, s1 nuclease mapping and primer extension to analyze transcripts derived from several genes in neurospora crassa mitochondria. the transcripts studied include those for cytochrome oxidase subunit iii, 17s rrna and an unidentified open reading frame. in all three cases, initial transcripts are long, include trna sequences, and are subsequently processed to generate the mature rnas. we find that endpoints of the most abundant transcripts generally coinci ...19852990893
structure of a neurospora rna polymerase i promoter defined by transcription in vitro with homologous extracts.a neurospora in vitro transcription system has been developed which specifically and efficiently initiates transcription of a cloned neurospora crassa ribosomal rna gene by rna polymerase i. the initiation site of transcription (both in vitro and in vivo) appears to be located about 850 bp from the 5' end of mature 17s rrna. however, the primary rrna transcripts are normally cleaved very rapidly at a site 120-125 nt from the 5' end in vitro and in vivo. the nucleotide sequence surrounding the in ...19852989792
nucleotide sequence of yeast gdh1 encoding nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase.the yeast gdh1 gene encodes nadp-dependent glutamate dehydrogenase. this gene was isolated by complementation of an escherichia coli glutamate auxotroph. nadp-dependent glutamate dehydrogenase was overproduced 6-10-fold in saccharomyces cerevisiae bearing gdh1 on a multicopy plasmid. the nucleotide sequence of the 1362-base pair coding region and 5' and 3' flanking sequences were determined. transcription start sites were located by s1 nuclease mapping. regulation of gdh1 was not maintained when ...19852989290
an equilibrium between distorted and undistorted dna in the adult chicken beta a-globin gene.we have used single strand specific nucleases to map dna distortion in the adult chicken beta a-globin gene. we have detected two structures of that kind and have mapped nuclease-cutting sites at one base resolution. one prominent site is centered at -190 relative to the rna capping site and is positioned at the center of a stretch of contiguous c residues. the second site is near the first intron/exon junction (+620) and appears as a series of discrete 1-base-long enzyme-cutting sites. based up ...19852989280
a water-soluble form of porin from the mitochondrial outer membrane of neurospora crassa. properties and relationship to the biosynthetic precursor form.mitochondrial porin, the outer membrane pore-forming protein, was isolated in the presence of detergents and converted into a water-soluble form. this water-soluble porin existed under nondenaturing conditions as a mixture of dimers and oligomers. the proportion of dimers increased with decreasing porin concentration during conversion. water-soluble porin inserted spontaneously into artificial bilayers as did detergent-solubilized porin. whereas the latter form had no specific requirements for t ...19852989279
cloning and characterization of the rdna repeat unit of podospora anserina.dna coding for ribosomal rna in podospora anserina has been cloned and was found as a tandemly repeated 8.3 kb sequence. the cloned rdna was characterized by restriction endonuclease mapping. the location of 5.8s, 18s and 28s rrna coding regions was established by dna-rna hybridization and s1 nuclease mapping. the organization of p. anserina rrna genes is similar to that of neurospora crassa and aspergillus nidulans. the rdna unit does not contain the sequence coding for 5s rna.19852987647
acetylglutamate kinase-acetylglutamyl-phosphate reductase complex of neurospora crassa. evidence for two polypeptides.mutations at the arg-6 locus in neurospora crassa are divided into two complementation groups (a and b) and a third noncomplementing group. there are many suppressible nonsense mutations among mutants in complementation group b and one in the noncomplementing group; no nonsense mutations exist among mutants in complementation group a (davis, r. h., and weiss, r. l. (1983) mol. gen. genet. 192, 46-50). we show here that the mutants are defective in either or both of two enzymes of arginine biosyn ...19852987210
influence of neurospora endonuclease on trenimon-induced structural chromosomal aberrations and sister-chromatid exchanges in human peripheral lymphocytes and chinese hamster ovary cells.human peripheral lymphocytes and chinese hamster ovary cells were treated in the g1 phase of the cell cycle with the trifunctional alkylating agent trenimon (trn) and post-treated with a single-strand specific endonuclease from neurospora crassa (ne). trn induces chromosomal aberrations of the chromatid type (ca) and sister-chromatid exchanges (sce). ne post-treatment leads to an elevation of the frequencies of ca but not of sces. this indicates that trn induced ca are the result of dna double-s ...19852985980
evidence for a common siderophore transport system but different siderophore receptors in neurospora crassa.uptake and competition experiments were performed with neurospora crassa and penicillium parvum by using 14c-labeled coprogen and 55fe-labeled ferrichrome-type siderophores. several siderophores of the ferrichrome family, such as ferrichrome, ferricrocin, ferrichrysin, and tetraglycyl-ferrichrome as well as the semisynthetic ferricrocin derivatives o-(phenyl-carbamoyl)-ferricrocin and o-(sulfanilyl-carbamoyl)-ferricrocin were taken up by n. crassa. the ferrichrome-type siderophores used vary in ...19852985545
concerted evolution of dispersed neurospora crassa 5s rna genes: pattern of sequence conservation between allelic and nonallelic genes.about 100 genes coding for 5s rna in neurospora crassa are dispersed throughout the genome (selker et al., cell 24:815-818, 1981; r. l. metzenberg, j. n. stevens, e. u. selker, and e. morzycka-wroblewska, manuscript in preparation). the majority of them correspond to the most abundant species (alpha) of 5s rna found in the cell. gene conversion, gene transposition, or both may be responsible for the maintenance of sequence homogeneity (concerted evolution) of alpha-type 5s genes. to explore thes ...19852984554
[detection and intracellular localization of 2 polyphosphate phosphohydrolases with different substrate specificities in the slime variant of neurospora crassa]. 19852982559
the neurospora mitochondrial genome: the region coding for the polycistronic cytochrome oxidase subunit i transcript is preceded by a transfer rna gene.we have sequenced a 682 bp fragment of neurospora crassa mitochondrial dna to complete the sequence between the gene for cytochrome b and the unassigned reading frame, urf u. the sequence contains a gene for a cysteine trna. the 5' end of the 6 kb polycistronic transcript of cytochrome c oxidase subunit 1 is immediately downstream from this trna. this shows that also in fungal mitochondria trnas can be used as processing signals, whereas palindromic sequences containing double pst i sites, also ...19852981709
loss of temperature compensation of circadian period length in the frq-9 mutant of neurospora crassa.a new circadian clock mutant of neurospora crassa has been isolated, whose most distinctive characteristic is the complete loss of temperature compensation of its period length. the q10 of the period length was found to be equal to about 2 in the temperature range from 18 degrees to 30 degrees c. the period length was also found to be dependent on the composition of the medium, including the nature and concentration of both the carbon source and the nitrogen source. although the rate of the cloc ...19862980965
phase shifting of the circadian conidiation rhythm in neurospora crassa by calmodulin antagonists.the effects of chemicals capable of antagonizing the functions of calmodulin, such as trifluoperazine, chlorpromazine, imipramine, alprenolol, w7, and w13, on the circadian conidiation rhythm of neurospora crassa were examined. trifluoperazine, at a 30-microm concentration, was most effective in shifting the phase of the conidiation rhythm and caused a maximum phase delay at circadian time (ct) 6 and maximum phase advance at ct 9. chlorpromazine was less effective than trifluoperazine, and a 300 ...19862980964
high-performance liquid chromatography of siderophores from fungi.a reversed-phase hplc separation of iron(iii) chelates of 16 representative fungal siderophores including ferrichromes, coprogens and triacetylfusarinine c was established in order to investigate siderophore production of fungi. for comparison purposes, the widely used bacterial siderophore ferrioxamine b was included. culture filtrates of the fungi penicillium resticulosum, fusarium dimerum, aspergillus fumigatus and neurospora crassa were quantitatively analyzed for the presence of known and u ...19882978959
ferricrocin functions as the main intracellular iron-storage compound in mycelia of neurospora crassa.neurospora crassa produces several structurally distinct siderophores: coprogen, ferricrocin, ferrichrome c and some minor unknown compounds. under conditions of iron starvation, desferricoprogen is the major extracellular siderophore whereas desferriferricrocin and desferriferichrome c are predominantly found intracellularly. mössbauer spectroscopic analyses revealed that coprogen-bound iron is rapidly released after uptake in mycelia of the wild-type n. crassa 74a. the major intracellular targ ...19882978956
dual roles for calcium ions in apical growth of neurospora crassa.we report initial attempts to define the role of ca2+ in the polarized extension of neurospora crassa. growth of the organism was diminished in media containing less than 1 mm-ca2+; extension was more severely impaired than biomass synthesis, resulting in the formation of stubby, bulbous hyphae, even of spherical cells. reduced extension and abnormal morphology were correlated with the loss of surface-bound ca2+, probably associated with the cell wall. intracellular ca2+ may be represented by ma ...19882978297
dna methylation and control of genome organization in neurospora crassa. 19882977770
defective myo-inositol-1-phosphate synthase production in an inositolless double mutant neurospora crassa strain.a slow growing inl+/- mutant was isolated from an inositol dependent (inl) neurospora crassa strain. the latter strain produces defective myo-inositol-1-phosphate synthase which has residual activity. inositol, similarly to that found in wild and inl mutant strains, represses the enzyme production in the inl+/- strain as well. withdrawing inositol from the medium results in derepression of the enzyme synthesis. derepression is hindered by cycloheximide. inl+/- character in the double mutant is b ...19882977674
chemical synthesis and expression of copper metallothionein gene of neurospora crassa.the gene coding for the neurospora crassa copper metallothionein (mt) was synthesized and inserted in the lacz' gene of puc18 plasmid to give the same translational reading frame as the latter gene. the mt-beta-galactosidase fused gene was expressed in escherichia coli to produce a fused protein in which the amino and carboxy termini of mt are linked to the beta-galactosidase through methionine residues. an mt derivative containing an extra homoserine residue at the carboxy terminus was prepared ...19882977386
blue light induces circadian rhythms in the bd mutant of neurospora: double mutants bd,wc-1 and bd,wc-2 are blind.this paper describes a new blue light effect for neurospora crassa, the photoinduction of circadian rhythms in the bd mutant. the wc-1 and wc-2 genes are necessary for this effect.19882977186
the amino acid sequence of ribonuclease n1, a guanine-specific ribonuclease from the fungus neurospora crassa.the complete amino acid sequence of ribonuclease n1 (rnase n1), a guanine-specific ribonuclease from a fungus, neurospora crassa, was determined by conventional protein sequencing, using peptide fragments obtained by tryptic digestion of cyanogen bromide-treated rnase n1 and by staphylococcus aureus v8 protease digestion of heat-denatured rnase n1. the results showed that the protein is composed of a single polypeptide chain of 104 amino acid residues cross-linked by two disulfide bonds and has ...19882977130
mating type response in neurospora crassa. early and transient changes in the patterns of protein synthesis in sexually stimulated mycelia.1. pulse labeling with [35s]-methionine, one-dimensional sds-polyacrylamide gel electrophoresis and fluorography were used to study the pattern of protein synthesis in neurospora crassa mycelia undergoing sexual development. 2. contact of sexually-competent mycelium with cells of the opposite mating type elicited a rapid and transient increase in the synthesis of two predominant proteins of 58 kda and 40 kda localized in the cytosol fraction. 3. marked changes in the pattern of protein synthesis ...19882977101
molecular organisation of the quinic acid utilization (qut) gene cluster in aspergillus nidulans.the functional integrity of the qutb gene (encoding quinate dehydrogenase) has been confirmed by transformation of a qutb mutant strain. the dna sequence of the contiguous genes qutd (quinate permease), qutb and qutg (function unknown) has been determined and analysed, together with that of qute (catabolic 3-dehydroquinase). the qutb sequence shows significant homology with the shikimate dehydrogenase function of the complex arom locus of aspergillus nidulans, and with the qa-3 quinate dehydroge ...19882976880
chitinase from neurospora crassa. 20102976112
molecular characterization of the mitochondrial dna of a new stopper mutant er-3 of neurospora crassa.an ethidium bromide-induced stopper mutant of neurospora crassa is characterized at the molecular level. the mutant has two populations of mitochondrial dna: a defective predominant mutant molecule and a basal level of the wild-type molecule. the aberrant dna resulted after a 25-kbp deletion from the wild-type mitochondrial chromosome, which included major genes such as cytb, co1 and oli2. the deletion endpoints are located in the second intron of the nd5 gene, and in a sequence 250 nucleotides ...19882976009
characterization of a mutation that causes overproduction of inositol in neurospora crassa.slow-growing (inl+/-) spontaneous mutants have been isolated from an inositol requiring (inl) strain of neurospora crassa that produces defective myo-inositol-1-phosphate synthase (mips), the enzyme responsible for the production of inositol-1-phosphate from glucose-6-phosphate. the defective enzyme has some residual activity. in the inl+/- strain the synthesis of the defective enzyme is enhanced, which enables the strain to grow slowly on minimal medium. the mutation (opi1) responsible for the ...19882975749
sequence analysis of mitochondrial dna from podospora anserina. pervasiveness of a class i intron in three separate genes.a 48 kb region of the 95 kb mitochondrial genome of podospora anserina has been mapped and sequenced (1 kb = 10(3) base-pairs). the dna sequence of the genes for nd2, 3, 4, atpase 6 and urfc are presented here. as in neurospora crassa, the nd2 and 3 genes consist of a unit separated by one taa stop codon. nd3, 4 and atpase 6 are interrupted by class i introns. all three introns are remarkably similar in the c-domain of their secondary structure, sufficient enough to designate them as new subgrou ...19882975708
metabolism of d-glucose in a wall-less mutant of neurospora crassa examined by 13c and 31p nuclear magnetic resonances: effects of insulin.13c nmr and 31p nmr have been used to investigate the metabolism of glucose by a wall-less strain of neurospora crassa (slime), grown in a supplemented nutritionally defined medium and harvested in the early stationary stage of growth. with d-[1-13c]- or d-[6-13c]glucose as substrates, the major metabolic products identified from 13c nmr spectra were [2-13c]ethanol, [3-13c]alanine, and c1- and c6-labeled trehalose. several observations suggested the existence of a substantial hexose monophosphat ...19882975509
luminescence from the carbon monoxide derivative of agaricus bispora tyrosinase.the luminescence of the co adduct of two isozymic tyrosinases isolated from agaricus bispora, an edible white mushroom, has been studied. at room temperature the emission appears as a single smooth peak centered at 530 nm with fwhm of 2700 cm-1 and a lifetime of 36 microseconds. the lifetime and wavelength of the emission are virtually unchanged on lowering the temperature from 298 to 77 degrees k. solvent composition affects the wavelength of emission minimally. the emission is quenched by oxyg ...19882975158
polyamine transport in neurospora crassa.polyamine transport in neurospora crassa is concentrative and energy dependent in a dilute buffer. the saturable systems governing the uptake of putrescine (km = 0.6 mm), spermidine (km = ca. 0.24 mm), and spermine (km = 0.07 mm) share components, as indicated by mutual inhibition among the polyamines. in addition, nonsaturable components prevail for putrescine and spermidine, particularly the former. radiolabeled substrates, once in the cell, are released only slowly, even if unlabeled polyamin ...19882975157
comparison of the orotidine 5'-monophosphate decarboxylase sequences of eight species.predicted amino acid sequences of the enzyme orotidine 5'-phosphate decarboxylase (ec 4.1.1.23) from eight different organisms are compared. the comparisons are made on the basis of primary structural differences, primary amino acid sequence, hydropathy profiles, and secondary structure predictions. the organisms compared are mus musculus, aspergillus nidulans, neurospora crassa, kluyveromyces lactis, saccharomyces cerevisiae, schizosaccharomyces pombe, escherichia coli, and salmonella typhimuri ...19882974823
presence of abnormal synaptonemal complexes in heterothallic species of neurospora.synaptonemal complex abnormalities are frequent in reconstructed meiotic prophase nuclei of neurospora crassa and neurospora intermedia. three kinds of synaptonemal complex anomalies were seen: lateral component splits, lateral component junctions, and multiple complexes. the anomalies apparently are formed during or after the pairing process, as they were not seen in the largely unpaired early zygotene chromosomes. their presence at all the other substages from mid-zygotene to late pachytene in ...19882974436
transcellular ionic currents studied by intracellular potential recordings in neurospora crassa hyphae. transfer of energy from proximal to apical cells.membrane potentials, input resistances, and electric coupling in the apical parts of n. crassa growing hyphae were recorded with the aid of intracellular microelectrodes. it was revealed that the apical cells were always depolarized by 10 to 30 mv as compared to the adjacent proximal cells. the septal pore maintained an electrical resistance of 4 to 6 m omega. the calculated values of the endogenous electrical current passing through the septal pore varied between 0.5 and 1 na. electrical isolat ...19882973993
[mechanism of photoregulation of carotenogenesis in neurospora crassa: the use of mutants]. 19882973407
binding of a 30-kda protein to the pyruvate kinase gene of neurospora crassa.extracts of a wild-type strain of neurospora crassa, electrophoresed on sds-polyacrylamide gels and electroblotted onto nitrocellulose sheets, were hybridized to an end-labelled pyruvate kinase (pk) gene fragment containing the 5' noncoding sequence and a large part of the coding region. a 30-kda protein was found to bind strongly to the pk gene dna, while binding weakly to plasmid puc12 dna and to total n. crassa dna. probing of blots with individual restriction fragments derived from the pk ge ...19882973333
method for identification of intracellular free flavin species in the photosensitive fungus neurospora crassa.establishing the relative intracellular proportions of flavins in neurospora crassa (and in other organisms) in vivo may be hampered by degradation of flavins after homogenization of the cells. the system described here allows separation and identification of intracellular free and bound flavins under conditions restrictive for the fad-degrading enzyme(s). a "protective buffer" containing 0.1 m citrate adjusted to ph 4.0 with k2hpo4, 5 mm atp, and 0.5 mm edta prevents fad from rapid enzymatic cl ...19882973261
structure of d-prephenyllactate. a carboxycyclohexadienyl metabolite from neurospora crassa.a novel natural product structurally related to prephenate and arogenate was isolated from a mutant of neurospora crassa. this d-beta-(1-carboxy-4-hydroxy-2,5-cyclohexadiene-1-yl)-lactic acid is herein given the trivial name of d-prephenyllactate. the new metabolite is even more acid labile than is prephenate and is quantitatively converted to phenyllactate at mildly acidic ph. the structure characterization of prephenyllactate was performed using spectroscopic techniques (ultraviolet, 1h nmr, 1 ...19882972718
cyclosporin a-binding protein (cyclophilin) of neurospora crassa. one gene codes for both the cytosolic and mitochondrial forms.cyclophilin (cyclosporin a-binding protein) has a dual localization in the mitochondria and in the cytosol of neurospora crassa. the two forms are encoded by a single gene which is transcribed into mrnas having different lengths and 5' termini (approximately 1 and 0.8 kilobases). the shorter mrna specifies the cytosolic protein consisting of 179 amino acids. the longer mrna is translated into a precursor polypeptide with an amino-terminal extension of 44 amino acids which is cleaved in two steps ...19882971658
translocation of a fragment of invertase across microsomal vesicles isolated from neurospora crassa requires the hydrolysis of a nucleoside triphosphate.the step which requires the hydrolysis of a nucleoside triphosphate for translocation of a protein across microsome was investigated by studying translocation uncoupled from translation using two truncated products of invertase: one product contains the first 262 amino acids of the secreted invertase (inv262); the other, the first 104 amino acids (inv104). the truncated products were translated from rna transcripts without a stop codon. it is demonstrated that the translated products contain an ...19882971655
isolation of genes encoding the neurospora vacuolar atpase. analysis of vma-1 encoding the 67-kda subunit reveals homology to other atpases.the vacuolar membrane of neurospora crassa contains a h+-translocating atpase composed of at least three subunits with approximate molecular weights of 70,000, 60,000, and 15,000. both genomic and cdna clones encoding the largest subunit, which appears to contain the active site of the enzyme, have been isolated and sequenced. the gene for this subunit, designated vma-1, contains six small introns (60-131 base pairs) and encodes a hydrophilic protein of 607 amino acids, mr 67,121. within the seq ...19882971651
x-ray-induced specific locus mutations in the ad-3 region of two-component heterokaryons of neurospora crassa. i. modification of the heterozygous effects of multilocus deletions covering the ad-3a or ad-3b loci.the basis for the reduced growth rates of heterokaryons between strains carrying nonallelic combinations of gene/point mutations (ad-3r) and multilocus deletion mutations (ad-3ir) has been investigated by a simple genetic test. the growth rates of forced 2-component heterokaryons (dikaryons) between multilocus deletion mutations were compared with forced 3-component heterokaryons (trikaryons) containing an ad-3ar ad-3br double mutant as their third component. since the third component has no gen ...19882971138
a specific insulin receptor and tyrosine kinase activity in the membranes of neurospora crassa.cells of the wall-less ("slime") strain of neurospora crassa possess specific high affinity insulin binding sites on their cell surface. 125i-labeled bound insulin was not displaced from these cells by insulin-like growth factor ii (igf-ii), and was only weakly displaced by igf-i and proinsulin. cross-linking of 125i-labeled insulin with n. crassa cells using disuccinimidyl suberate resulted in the labeling of a single band of ca. 67 kda m.w. on a polyacrylamide gel. two proteins of ca. 66 and 5 ...19882970849
genetic analysis of transformation in a microconidiating strain of neurospora crassa.we have characterized neurospora crassa transformants obtained with plasmid pdv1001 bearing the cloned catabolic dehydroquinase (qa-2+) gene (hughes et al. 1983a) and fluffy 268 host strain producing only uninucleate microconidia allowing to isolate individual transformation products. the percentage of transformed nuclei in the mycelium and their stability were determined by genetic analysis of microconidia produced on selective or non-selective medium. about half of the transformants originatin ...19852970312
molecular analysis of a neurospora crassa gene expressed during conidiation.the asexual developmental pathway in the life cycle of the filamentous fungus neurospora crassa culminates in the formation of spores called conidia. several clones of genomic neurospora dna have been isolated that correspond to mrna species expressed during conidiation and not during mycelial growth (v. berlin and c. yanofsky, mol. cell. biol. 5:849-855, 1985). in this paper we describe the characterization of one of these clones, named pcon-10a. this clone contains two genes, con-10 and con-13 ...19882970007
growth regulation by gtp. regulation of nucleotide pools in neurospora by nitrogen and sulfur control systems.purine nucleotide pools in the fungus neurospora crassa decline in response to carbon, nitrogen, or sulfur deprivation. there is, in addition, a decline in gtp/atp ratios on nitrogen or sulfur deprivation in wild type. the gtp/atp decline is missing on nitrogen deprivation of the nitrogen control mutant, nit-2, and on sulfur deprivation of the sulfur control mutant, cys-3. the nit-2 mutant also shows elevated utp pools on nitrogen deprivation when compared with similarly treated wild type. six-h ...19882969890
a neurospora crassa heat-shocked cell lysate translates homologous and heterologous messenger rna efficiently, without preference for heat shock messages.cell-free protein synthesis systems were prepared from normally-grown (n-lysate) and heat-shocked (hs-lysate) neurospora crassa mycelium. although both lysates translated homologous mrna, the hs-lysate was more active, yielding a higher incorporation of [35s]-methionine into hot tca-insoluble material and a vastly superior protein synthesis profile. the optimal temperature for translation by both lysates was 21 degrees c; the hs-lysate did not translate heat-shock mrna preferentially at any temp ...19882969781
relationship of histidine sensitivity to dna damage and stress induced responses in mutagen sensitive mutants of neurospora crassa.previous work in other laboratories has shown that several mutagen sensitive mutants of neurospora crassa are extremely sensitive to low levels of histidine in the culture medium. we have shown that wild type neurospora accumulates nicks or breaks in the dna in the presence of histidine. the number of nicks accumulating in histidine sensitive mutants is found to increase in relation to their sensitivity to histidine. although these nicks can be repaired by both wild type and histidine sensitive ...19882969780
ethanol and carbon-source starvation enhance the accumulation of hsp80 in neurospora crassa.in neurospora crassa, heat shock results in the induction of 9 to 11 heat shock proteins (hsp), of which hsp80 is the most abundant and the first to be synthesized. the induction of hsp80 was investigated during normal growth (2% sucrose) and under sucrose starvation. transfer of mycelium to a medium supplemented with ethanol stimulated the synthesis of hsp80, even at the normal growth temperature of 28 degrees c. it was also synthesized under carbon starvation conditions, where the medium was s ...19882969770
the overexpression, purification and complete amino acid sequence of chorismate synthase from escherichia coli k12 and its comparison with the enzyme from neurospora crassa.the enzyme chorismate synthase was purified in milligram quantities from an overproducing strain of escherichia coli. the amino acid sequence was deduced from the nucleotide sequence of the aroc gene and confirmed by determining the n-terminal amino acid sequence of the purified enzyme. the complete polypeptide chain consists of 357 amino acid residues and has a calculated subunit mr of 38,183. cross-linking and gel-filtration experiments show that the enzyme is tetrameric. an improved purificat ...19882969724
induction of multiple germ tubes in neurospora crassa by antitubulin agents.the antitubulin fungicide benomyl suppressed the linear growth of neurospora crassa wild type strain st. lawrence 74 at micromolar concentrations. the rate of germination of macroconidia was not affected. macroconidia exposed to 1.7 microm benomyl for 5 h formed multiple germ tubes. when germlings incubated for 4 h were exposed to 1.7 microm benomyl for 3 h, their germ tube stopped growing, swelled and emitted several branches. normal linear growth was restored after removal of the fungicide. li ...19882969337
isolation and characterization of non-neuronal enolase (nne) from neurospora crassa and comparison with neuron specific enolase isolated from neuroblastoma cell line ng108.enolase is a vital enzyme of the glycolytic pathway. it exists mainly in two forms, non-neuronal enolase (nne) and neuron specific enolase (nse). neurospora crassa, a filamentous fungus, was used as the source of pure nne, and by using deae-cellulose and a sephadex g-150 column chromatography highly purified enzyme (20.4 fold purification with 54.7 percent recovery) was obtained. the development profile of the enzyme shows a peak value after 90 hours of mycelial growth from conidia of n. crassa. ...19872969242
deoxyribonucleoside triphosphate pools in neurospora crassa: effects of histidine and hydroxyurea.an effective hplc method for detecting deoxyribonucleoside triphosphates in hyphae from the fungus neurospora crassa has been developed. in rapidly growing cells the nucleotide levels vary from 11.8 pmoles/micrograms dna for dgtp to 24.2 pmoles/micrograms dna for dttp. these levels fall by approximately one half in stationary-phase cultures but the ratio of each pool to dgtp remains the same. the dntp pools in conidia are at least 5-fold lower than in rapidly growing cells. the pool sizes are th ...20132969078
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