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two n-hydroxylaminopurines are highly mutagenic in the ad-3 forward-mutation test in growing cultures of heterokaryon 12 of neurospora crassa.3 purine analogs were tested for their mutagenic activities in the ad-3 forward-mutation test in heterokaryon 12 (h-12) of neurospora crassa. in growing cultures of h-12, the n-hydroxylaminopurines 2-amino-6-n-hydroxylaminopurine (aha) and 6-n-hydroxylaminopurine (hap) are potent and strong mutagens, respectively, whereas 2-aminopurine (ap) is a weak mutagen. aha and hap are about equally mutagenic at low doses, but aha is more mutagenic than hap at high doses. despite their potent mutagenicity ...19872950320
metabolic utilization of 57fe-labeled coprogen in neurospora crassa. an in vivo mössbauer study.mössbauer spectra of whole cells of neurospora crassa arg-5 ota aga (a siderophore-free mutant) show that the siderophore coprogen is accumulated inside the cell as an entity. 57fe from 57fe-labeled coprogen is slowly removed from the complex (45% in 27 h). the rate of removal depends on the degree of iron starvation of the cells. the distribution of 55fe from [55fe]coprogen in vacuoles, membranes, and cytoplasm has been also determined. from this it is clear that coprogen is accumulated in the ...19872951253
role of nitrogen in the photoinduction of protoperithecia and carotenoids in neurospora crassa.nitrogen, as kno3 or nh4no3, can inhibit the photoinduction of protoperithecia in neurospora crassa when present in the medium at a high concentration but does not inhibit the photoinduction of carotenoids. the point at which the presence of high nitrogen levels is no longer inhibitory is 5 h after illumination.198724232879
intracellular and extracellular cyclic nucleotides in wild-type and white collar mutant strains of neurospora crassa: temperature dependent efflux of cyclic amp from mycelia.cyclic amp and cyclic gmp were released into the growth medium of mycelia of neurospora crassa wild-type strains st.l.74a and em5297a and by white collar-1 and white collar-2 mutant strains. after growth for 6 days at 18 degrees c, there were 2.19 (st.l.74a), 5.83 (em5297a), 1.38 (white collar-1), and 1.10 (white collar-2) nanomoles of cyclic amp per gram dry weight of mycelia in the growth medium. these values corresponded to concentrations of cyclic amp of between approximately 10 and 50 nanom ...198716665253
three class i introns in the nd4l/nd5 transcriptional unit of neurospora crassa mitochondria.the overlapping nd4l and nd5 genes of neurospora crassa mitochondria are interrupted by one and two intervening sequences, respectively, of about 1,490, 1,408 and 1,135 bp in length. all three intervening sequences are class i introns and as such have the potential to fold into the conserved secondary structure that has been proposed for the majority of fungal mitochondrial introns. they contain long open reading frames (orfs; from 306 to 425 codons long) that are continuous and in frame with th ...19872953954
a hexameric form of the neurospora crassa plasma membrane h+-atpase.as isolated by our recently developed large-scale procedure, the neurospora plasma membrane h+-atpase exists as a homogeneous, oligomeric complex of 105,000-da monomers with a molecular mass equivalent to a spherical protein of about 1 million da, as judged by its behavior during chromatography on calibrated columns of sepharose cl-6b and cl-4b. treatment of this complex with the nonionic detergent, tween 20, followed by sepharose column chromatography in the presence of this detergent produces ...19872880563
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
isolation and characterization of a dna-uptake-stimulating protein from the culture medium of neurospora crassa slime strain.a protein fraction was purified to homogeneity from the culture medium of the wall-less (slime) strain of neurospora crassa (fgsc 1118), which proved to be identical with dna-uptake-stimulating factor (designated dusf), which has been described earlier [schablik, m. and szabó, g. (1981) fems microbiol. lett. 10, 395-397]. the quantity of dusf is measured by the amount of [3h]dna uptake by neurospora cells at standard conditions. its relative molecular mass was 230,000. it has an isoelectric poin ...19872949969
monophenol monooxygenase from neurospora crassa. 19873037257
applicability of the equations of freundlich and langmuir to the adsorption of the azo dye procion scarlet on paramorphic colonies of neurospora crassa.experiments on the adsorption of procion scarlet mx-g by normal hyphae and by paramorphic colonies of neurospora crassa were performed at ph 2.5, 4.5 and 6.5 at 30 degrees c. the measured adsorption isotherms were evaluated by the freundlich and langmuir equations. the removal of dye was most effective at ph 2.5 and more dye was adsorbed per unit mass of cells in the paramorphic cultures than in the normal hyphae. the statistical tests showed langmuir's equation to give a better fit to the adsor ...19872968127
possible link between circadian rhythm and heat shock response in neurospora crassa.3-h pulses of elevated temperatures (30 degrees c, 35 degrees c, 40 degrees c) phase shift the circadian conidiation rhythm of neurospora crassa. the phase and amplitude of the phase response curves (prc) were measured in wild type (frq+) and frequency mutants (frq 1, frq 7). the dose dependence of the phase shifts was compared to the dose dependence of total protein synthesis inhibition and heat shock protein induction in the three strains. all processes showed an almost linear dependence on te ...19872963703
regulation of the qa gene cluster of neurospora crassa. 19872961304
differential synthesis and replication of dna in the neurospora crassa slime mutant versus normal cells: role of carcinogens.small quantities of carcinogens, dl-ethionine, thiotepa, actinomycin d, and 1-(2-chloroethyl-3-cyclohexyl)-1-nitrosourea (ccnu) stimulated in vitro deoxyribonucleic acid (dna) synthesis of the slime mutant of neurospora crassa, while there was practically no effect on the dna from the normal wild type 74a strain. all of these compounds caused increased strand separation in the mutant dna of n. crassa, but no separation of normal dna strands. the growth (in vivo tests) of the n. crassa slime muta ...19872959890
isolation and regulation of expression of the neurospora crassa copper metallothionein gene.the n. crassa cumt gene has been cloned and its nucleotide sequence determined. to this end an mt specific undecanucleotide was synthesized and used for cdna synthesis with enriched mt mrna as a template. sequence analysis of the cdna obtained allowed the synthesis of a unique 21mer which was used as a hybridization probe to screen a genomic dna library of n. crassa. several positive clones were isolated and subjected to restriction and sequence analysis. in agreement with the published amino ac ...19872959528
isolation and characterization of an extracellular lipase from the conidia of neurospora crassa.a triacylglycerol lipase (ec 3.1.1.3) from the conidia of neurospora crassa was purified and characterized. the enzyme was purified by sephadex g-100 column chromatography. homogeneity was checked by page, and isoelectric focusing gave a single band corresponding to a pi of 6.4. the enzyme had an apparent mr 54000 +/- 1000 as determined by gel filtration. sds-page gave a single band of mr 27000, suggesting the presence of two identical subunits. this lipase preferred triglycerides with c16- and ...19872958597
the arom multifunctional enzyme from neurospora crassa. 19872955200
analysis of mutational lesions of acetate metabolism in neurospora crassa by 13c nuclear magnetic resonance.the adaptation of neurospora crassa mycelium to growth on acetate as the sole carbon source was examined by using 13c nuclear magnetic resonance. extracts were examined by nuclear magnetic resonance at various times after transfer of the mycelium from medium containing sucrose to medium containing [2-13c]acetate as the sole carbon source. the label was initially seen to enter the alanine, glutamate, and glutamine pools, and after 6 h 13c-enriched trehalose was evident, indicating that gluconeoge ...19872947898
transformation of aspergillus niger using the homologous orotidine-5'-phosphate-decarboxylase gene.a homologous transformation system for the filamentous fungus aspergillus niger has been developed, based on the orotidine-5'-phosphate-decarboxylase gene. a. niger pyr- mutants have been selected from 5-fluoro-orotic acid resistant mutants. these mutants were found to comprise two complementation groups, pyra and pyrb. the a. niger omp-decarboxylase gene was isolated from a gene library by heterologous hybridization with the neurospora crassa pyr4 gene. the cloned gene is capable to transform a ...19872836081
identification and isolation of a putative permease gene in the quinic acid utilization (qut) gene cluster of aspergillus nidulans.mutations in the qutd gene of aspergillus nidulans cause the loss of ability to grow upon quinic acid as sole carbon source in media at normal ph 6.5 and failure to induce three enzyme activities specifically required for metabolism to protochatechuic acid. all 9 qutd mutants recovered are recessive and have been found to be ph sensitive, growing strongly on quinic acid media at ph 3.5 and producing significant induced enzyme activities. these properties are consistent with the hypothesis that t ...19872835177
neurospora crassa nuclear genome contains analogy of saccharomyces cerevisiae genes for ribosomal rna processing.neurospora crassa wild type genome shows dna sequences which are homologous to the sequences present in the rrna processing genes of the yeast saccharomyces cerevisiae. five such processing genes from yeast, viz., rna1 through rna5, cloned in plasmid pbr322 were transformed in escherichia coli strain le392. southern blots containing dnas from these clones were restricted with several restriction endonucleases along with dnas from lambda phage, rice (plant) and neuroblastoma (animal), were hybrid ...19872835180
gamma-ray-sensitive mutants of neurospora crassa with characteristics analogous to ataxia telangiectasia cell lines.well characterized gamma-ray sensitive mutants of the fungus neurospora crassa have been screened for characteristics analogous to those of cell lines derived from humans with the genetic disease, ataxia telangiectasia (at). two neurospora mutants, uvs-6 and mus-9, show the at cell line characteristics of gamma-ray and bleomycin sensitivity, and little or no repression of dna synthesis following treatment with these agents. normal human or neurospora cells show an extensive biphasic dna synthesi ...19872434849
particular rna primer from growth medium differentially stimulates in vitro dna synthesis and in vivo cell growth of neurospora crassa and its slime mutant.purine rich small "rna-primer" molecules (about 10-12 nucleotides), secreted into the growth medium of 3-h germinated conidia of n. crassa, strongly stimulated a concentration-dependent in vitro dna synthesis of n. crassa slime mutant as well as dnas from the human cancer cells but did not affect that from normal cells. these "rna-primer" molecules stimulated also in vivo cell growth of n. crassa slime mutant, but not of the n. crassa wild type. our studies suggest that dnas from the slime mutan ...19872452704
the molybdenum iron-sulphur protein from desulfovibrio gigas as a form of aldehyde oxidase.the molybdenum iron-sulphur protein originally isolated from desulfovibrio gigas by moura, xavier, bruschi, le gall, hall & cammack [(1976) biochem. biophys. res. commun. 72, 782-789] has been further investigated by e.p.r. spectroscopy of molybdenum(v). the signal obtained on extended reduction of the protein with sodium dithionite has been shown, by studies at 9 and 35 hgz in 1h2o and 2h2o and computer simulations, to have parameters corresponding to those of the slow signal from the inactive ...19872821990
purification and characterization of an endo-exonuclease from saccharomyces cerevisiae that is influenced by the rad52 gene.an endo-exonuclease has been purified from logarithmically growing cells of the yeast saccharomyces cerevisiae. identification and purification of this nuclease was facilitated by its being precipitable with an antibody raised against a previously described neurospora crassa endo-exonuclease (resnick, m. a., chow, t. y.-k. nitiss, j., and game, j. c. (1984) cold spring harbor symp. quant. biol. 49, 639-649 and t. y.-k. chow and m. a. resnick (1988) mol. gen. genet., in press). the enzyme which w ...19872826428
nitrobacter winogradskyi cytochrome a1c1 is an iron-sulfur molybdoenzyme having hemes a and c.cytochrome a1c1 (nitrite-cytochrome c oxidoreductase) purified from nitrobacter winogradskyi (formerly n. agilis) contained molybdenum, non-heme iron, and acid-labile sulfur in addition to hemes a and c; it contained 1 mol of heme a, 4-5 g atoms of non-heme iron, 2-5 g atoms of acid-labile sulfur, and 1-2 g atoms of molybdenum per mol of heme c, but did not contain copper. the fluorescence spectra of the molybdenum cofactor derivative prepared from cytochrome a1c1 were very similar to those of t ...19872828343
development of a homologous transformation system for aspergillus niger based on the pyrg gene.the development of a homologous transformation system for aspergillus niger is described. the system is based on the use of an orotidine-5'-phosphate decarboxylase deficient mutant (pyrg) and a vector, pab4-1, which contains the functional a. niger pyrg gene as a selection marker. transformation of the a. niger pyrg mutant with pab4-1 resulted in the appearance of stable pyr+ transformants at a frequency of 40 transformants per microgram of dna. in 90% of these transformants integration had occu ...19873472035
transformation of aspergillus oryzae using the a. niger pyrg gene.a transformation system for aspergillus oryzae based on the orotidine-5'-phosphate decarboxylase gene (pyrg) was developed. transformation frequencies of up to 16 transformants per microgram of dna were obtained with the vector pab4-1, which carries the pyrg gene of a. niger. southern blotting analysis showed that vector dna sequences were integrated into the chromosomal dna, in various copy numbers and presumably at different sites. efficient cotransformation of an unselectable gene was also sh ...19873481025
mitochondrial binding of a protein affected in mutants resistant to the microtubule inhibitor podophyllotoxin.specific antibodies to a protein p1 mr approximately equal to 63,000) from chinese hamster ovary cells, which is affected in mutants resistant to the microtubule inhibitor, podophyllotoxin, and behaves like a microtubule-related protein by certain criteria [14], have been raised. the antibody reacts specifically with the p1 protein in one- and two-dimensional immunoblots, and a cross-reacting protein of similar molecular mass and electrophoretic mobility is also found in cells from various verte ...19873319627
cloning, mapping and molecular analysis of the pyrg (orotidine-5'-phosphate decarboxylase) gene of aspergillus nidulans.we have modified the transformation procedures of ballance et al. [biochem. biophys. res. commun. 112 (1983) 284-289] to give increased rates of transformation in aspergillus nidulans. with the modified procedures we have been able to complement pyrg89, a mutation in the orotidine-5'-phosphate decarboxylase gene of a. nidulans, by transformation with a library of wild-type (wt) sequences in pbr329. we have recovered, by marker rescue from one such transformant, a plasmid (pjr15) that carries an ...19873328733
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
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
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
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
biosynthesis of thiamin. different biosynthetic routes of the thiazole moiety of thiamin in aerobic organisms and anaerobic organisms.the nitrogen atom of glycine was incorporated into the thiazole moiety of thiamin in the aerobic microorganisms bacillus subtilis, pseudomonas putida, saccharomyces cerevisiae, mucor racemosus, neurospora crassa, and emericella nidulans. it was not incorporated in the case of the facultative anaerobic microorganisms escherichia coli and enterobacter aerogenes, which, however, did incorporate the nitrogen atom of tyrosine. these results show that aerobic microorganisms and facultative anaerobic m ...19873566774
complete amino-acid sequence of a functional unit from a molluscan hemocyanin (helix pomatia).from the beta c-hemocyanin (beta c-hc) of the vineyard snail, helix pomatia, the functional unit d (mr approximately equal to 50,000-55,000) was isolated by limited proteolysis and gel chromatography. a small quantity of functional unit d was obtained intact, but the major part in the form of two peptides (mr approximately equal to 43,000 and 10,000, respectively) connected by a disulfide bridge. after reduction and carboxymethylation, these were separated from each other and cleaved by conventi ...19873620107
a long polypyrimidine/polypurine tract induces an altered dna conformation on the 3' coding region of the adjacent myosin heavy chain gene.a long (147 base pairs), natural a.t rich polypyrimidine/polypurine tract has been found 55 base pairs downstream of a chicken embryonic myosin heavy chain (mhc) gene. analysis at the nucleotide level of nicks induced by s1 and neurospora crassa nucleases indicate that this long interrupted polypyrimidine/polypurine tract exists in an alternate dna structure in vitro at ph 4.5 and ph 7.5 in both supercoiled and linear plasmid dna. the polypyrimidine/polypurine tract induces this alternate struct ...19873671071
extraction and purification of molybdenum cofactor from milk xanthine oxidase.molybdenum cofactor (mocofactor) is extracted efficiently, free of impurities and in high concentrations, by acid treatment of xanthine oxidase and subsequent incubation of the precipitate with phosphate buffer containing edta, molybdate and oxygen. it is suggested that cofactor is bound to the enzyme via hydrophobic forces as well as via an oxygen-sensitive mechanism. upon extraction, the capability to complement the apo nitrate reductase of neurospora crassa nit-1 can be conserved only in the ...19873691496
regulation of synthesis and secretion of acid and alkaline phosphatases in neurospora crassa.we show that n. crassa represses the production of acid phosphatase at ph higher than 8.0, irrespective of the carbon source used, whereas production was stimulated by sucrose at slightly acidic ph. the same profile of acid phosphatase production was observed in the pho-2a, pho-3a, nuc-1a, nuc-2a and pregc mutant strains. we also show that acid phosphatase synthesized by the pregc mutant strain grown on high phosphate medium has pronounced differences when compared to the enzyme synthesized by t ...19872967123
fungal small nuclear ribonucleoproteins share properties with plant and vertebrate u-snrnps.snrnas with properties closely related to those of the major vertebrate u-snrnas are present in the fungi aspergillus nidulans, neurospora crassa and schizosaccharomyces pombe. these rnas possess a tri-methyl guanosine cap structure and a subset cross-hybridizes with human u1 and u2 clones. in the form of snrnps, snrnas from these fungi as well as from saccharomyces cerevisiae and pea plants are immunoprecipitated by human and anti-sm or anti-(u1)rnp autoimmune antibodies. on micro-injection int ...19872953599
purification and characterization of arginase from neurospora crassa.we have purified an enzymatically active form of arginase from a wild-type strain of neurospora crassa to homogeneity. the enzyme has a subunit molecular weight of 38,300 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. the native protein migrated as a hexamer during gel-filtration chromatography with an apparent molecular weight of 266,000. the enzyme exhibited hyperbolic kinetics at ph 9.5 with an apparent km for arginine of 131 mm. antiserum was prepared against the ...19872953715
the neurospora crassa metallothionein gene. regulation of expression and chromosomal location.the promoter region of the neurospora crassa metallothionein gene contains no sequences which are similar to the mammalian or the yeast metal responsive elements (münger, k., germann, u. a., and lerch, k. (1985) embo j. 4, 2665-2668). we therefore studied the regulation of expression of the n. crassa metallothionein gene in response to different metal ions (cu2+, cd2+, zn2+, co2+, and ni2+) by northern analysis. only copper led to the induction of metallothionein mrna. in n. crassa cultures inoc ...19872953720
ornithine decarboxylase from neurospora crassa. purification, characterization, and regulation by inactivation.ornithine decarboxylase, a highly regulated enzyme of the polyamine pathway, was purified 670-fold from mycelia of neurospora crassa that were highly augmented for enzyme activity. the enzyme is significantly different from those reported from three other lower eucaryotic organisms: saccharomyces cerevisiae, physarum polycephalum, and tetrahymena pyriformis. instead, the enzyme closely resembles the enzymes from mammals. the mr = 110,000 enzyme is a dimer of 53,000 da subunits, with a specific a ...19872953728
complementation of area- regulatory gene mutations of aspergillus nidulans by the heterologous regulatory gene nit-2 of neurospora crassa.loss-of-function mutations in the regulatory gene area of aspergillus nidulans prevent the utilization of a wide variety of nitrogen sources. the phenotypes of nit-2 mutants of neurospora crassa suggest that this gene may be analogous to the area gene. transformation has been used to introduce a plasmid containing the nit-2 gene into a. nidulans. the nit-2 gene of neurospora complemented mutations in the area gene, restoring the ability to use a variety of nitrogen sources. this indicated that t ...19872954160
nucleotide sequence and characterization of the pyrf operon of escherichia coli k12.the pyrf gene of escherichia coli k12, which encodes the pyrimidine biosynthetic enzyme orotidine-5'-monophosphate (omp) decarboxylase, is part of an operon that includes a downstream gene designated orff. the orff gene product is a small polypeptide of unknown function. the nucleotide sequence of a 1549-base pair chromosomal fragment containing this operon was determined. an open reading frame capable of encoding the 27-kda omp decarboxylase subunit was identified and shown to be the pyrf struc ...19872956254
a portable signal causing faithful dna methylation de novo in neurospora crassa.methylation of cytosine residues in eukaryotic dna is common, but poorly understood. typically several percent of the cytosines are methylated; however, it is unclear what governs which sequences eventually become modified. neurospora crassa dna containing the "zeta-eta" (zeta-eta) region, which is a region of unusually heavy methylation, was tested for its ability to direct dna methylation de novo. dna stripped of its methylation by propagation in escherichia coli was reintroduced into neurospo ...19872958937
luminescence emission from the cu(i)-thiolate complex in metallothioneins.the luminescence emission properties of cu-metallothioneins (from neurospora crassa, agaricus bisporus and livers of bedlington terriers affected by copper toxicosis) as well as of (cu,zn)-metallothionein from bovine fetal liver are reported. upon excitation in the u.v., these proteins emit a largely red-shifted luminescence with a maximum at 565 nm attributable to the cu(i)-thiolate chromophores of the proteins. differences in the shapes of the spectra and the emission intensity are observed wi ...19872959510
orientation of enzymic domains in tryptophan synthase of neurospora crassa: an immunoblot analysis of trp3 mutant products.extracts of 52 trp3 mutants of neurospora crassa were tested for the presence of serologically cross-reacting material by the method of electrophoretic blot analysis. the test antigen was obtained by excision of lightly stained bands of denatured pure tryptophan synthase after sds-polyacrylamide gel electrophoresis. rabbit antisera raised against this antigen neutralized and precipitated native tryptophan synthase. of the 52 strains, 19 exhibited banding patterns similar to wild type on electrop ...19872959839
molecular genetic analysis of the pyr-4 gene of neurospora crassa.by means of s1 nuclease mapping, one transcription origin and three termini are identified for the pyr-4 gene of neurospora crassa, the same origin being used also by escherichia coli on the cloned gene. translation of the clone in mini-cells gives a 50,000 dalton gene product, the same size as that determined for the neurospora native enzyme. putative caat and tata boxes, and upstream and downstream potential secondary structures, are identified.19872959843
mitochondrial porin of neurospora crassa: cdna cloning, in vitro expression and import into mitochondria.cdna encoding porin of neurospora crassa, the major protein component of the outer mitochondrial membrane, was isolated and the nucleotide sequence was determined. the deduced protein sequence consists of 283 amino acids (29,979 daltons) and shows sequence homology of around 43% to yeast porin; however, no significant homology to bacterial porins was apparent. according to secondary structure predictions, mitochondrial porin consists mainly of membrane-spanning sided beta-sheets. porin was effic ...19872960519
role of siderophores in iron storage in spores of neurospora crassa and aspergillus ochraceus.spores of neurospora crassa 74a are lacking in ferritinlike iron pools, as demonstrated by mössbauer spectroscopic analysis. the cyclic hexapeptide siderophore ferricrocin constituted 47% of the total iron content in spores. after germination and growth, the ferricrocin iron pool disappeared, indicating that the metal was utilized. in spores of aspergillus ochraceus, 74% of the total iron content was bound by ferrichrome-type siderophores. siderophores may function as iron storage forms in funga ...19872960664
secretory protein translocation in a neurospora crassa in vitro system. hydrolysis of a nucleoside triphosphate is required for posttranslational translocation.an in vitro translocation system has been reconstituted with subcellular fractions from the cell wall-less mutant of neurospora crassa (fz;sg;os-1). prepro alpha factor and invertase, secretory proteins from yeast, were faithfully translocated and glycosylated by neurospora microsomes when presence cotranslationally in the neurospora translation system. when presence cotranslationally in the neurospora translation system, microsomes from canine pancreas(crm) could also translocate and glycosylat ...19872960680
isolation and characterization of a neurospora crassa ribosomal protein gene homologous to cyh2 of yeast.we have isolated and characterized a neurospora crassa gene homologous to the yeast cyh2 gene encoding l29, a cycloheximide sensitivity-conferring protein of the cytoplasmic ribosome. the cloned neurospora gene was isolated by cross-hybridization to cyh2. it was sequenced from both cdna and genomic clones. the coding region is interrupted by seven intervening sequences. its deduced amino acid sequence shows 70% homology to that of yeast ribosomal protein l29 and 60% homology to that of mammalian ...19872960953
human liver cdna clones encoding proteolipid subunit 9 of the mitochondrial atpase complex.clones encoding the proteolipid subunit 9 of the mitochondrial atpase complex have been isolated from a lambda gt10 library of human liver cdna sequences, using a probe of neurospora crassa cdna encoding subunit 9. from nucleotide sequence analysis it is concluded that the amino acid sequence of mature human subunit 9 is identical to that of its bovine counterpart. by comparing the sequence of two cdna clones (denoted human 1 and 2) to those of two bovine cdna clones (denoted p1 and p2) recently ...19872883974
studies on nitrate reductase of clostridium perfringens. iv. identification of metals, molybdenum cofactor, and iron-sulfur cluster.nitrate reductase of clostridium perfringens was purified by an improved method using immuno-affinity chromatography. the purified preparation contained mo, fe, and acid-labile sulfide; the mo content was 1 mol per mol and the fe 3.7 mol per mol of the enzyme. the inactive enzyme obtained from cells grown in the presence of tungstate did not hold mo but contained 1 mol of w. the content of fe was not increased. the presence of molybdenum cofactor in the nitrate reductase was indicated by the for ...19872884214
mutation of a conserved glycine residue modifies the vanadate sensitivity of the plasma membrane h+-atpase from schizosaccharomyces pombe.the structural gene pma+1 for the h+-atpase from the fission yeast schizosaccharomyces pombe has been isolated and sequenced. the intron-less gene encodes for a protein of mr = 99,769 which is 75% homologous to those of saccharomyces cerevisiae and neurospora crassa. the s. pombe pma+1 gene complements not only s. pombe pma-1-1 but also s. cerevisiae pma-1-4 mutants selected for in vitro vanadate-resistant atpase activity. the sequence of the s. pombe mutant pma-1-1 allele reveals that the glyci ...19872891694
clathrin coated vesicles in neurospora crassa.electropherograms of neurospora crassa homogenates showed a polypeptide with a mobility slightly lower than that of a standard sample of clathrin (from bovine brain). subcellular fractionation of the homogenate resulted in a 20-fold enrichment of the putative n. crassa clathrin in the microsomal fraction. further fractionation of the microsomal fraction by glass bead permeation chromatography yielded a fraction enriched about 150-fold relative to the homogenate. coated vesicles (42.5 +/- 2.5 nm ...19872892127
involvement of chla, e, m, and n loci in escherichia coli molybdopterin biosynthesis.all molybdenum enzymes except nitrogenase contain a common molybdenum cofactor, whose organic moiety is a novel pterin called molybdopterin (mpt). to assist in elucidating the biosynthetic pathway of mpt, two mpt-deficient mutants of escherichia coli k-12 were isolated. they lacked activities of the molybdenum enzymes nitrate reductase and formate dehydrogenase, did not reconstitute apo nitrate reductase from a neurospora crassa nit-1 strain, and did not yield form a, a derivative of mpt. by p1 ...19872947896
complex iii from mitochondria on neurospora crassa: purification, characterization, and resolution. 1987213696
effect of carbon source on enzymes involved in glycerol metabolism in neurospora crassa.specific activities of eight enzymes involved in glycerol metabolism were determined in crude extracts of three strains of neurospora crassa after growth on six different carbon sources. one of the strains was wild type, which grew poorly on glycerol as sole carbon source; the other two were mutant strains which were efficient glycerol utilizers. a possible basis for this greater efficiency of glycerol utilization was catabolite repression of glyceraldehyde kinase by glycerol in wild type, and t ...1987211971
influence of the carbon source on glycerol kinase activity in neurospora crassa.the level of glycerol kinase activity in neurospora crassa was shown to change in response to resuspension of sucrose-grown mycelia in fresh medium containing a new carbon source: the magnitude of the change depended on the new carbon source provided. certain carbon sources, such as glucose and fructose, inhibited the small increase that occurred in the absence of any carbon source. others, and in particular deoxyribose, galactose, glycerol and ribose, greatly enhanced this increase. the activit ...1987174743
an altered invertase in the cot-2 mutant of neurospora crassa.because the cot-2 and inv loci of neurospora crassa are closely linked, the invertase from the morphological mutant, cot-2, was examined. the cot-2 strains produce an invertase with altered heat sensitivity, km, and ratio of heavy to light forms. the cellular localization of cot-2 invertase is different from that of the wild type. there were no observable changes in the energy of activation or the ph optimum of cot-2 invertase, and some of the differences detected were not apparent under culture ...1986239095
mode of action and properties of xylanase and beta-xylosidase from neurospora crassa.extracellular beta-xylosidase (1,4-beta-d-xylan xylohydrolase, ec 3.2.1.37) from culture filtrates of neurospora crassa was purified to homogeneity by preparative isoelectric focusing followed by gel electrophoresis. the molecular weight of the purified xylosidase was 83,000 d and the k(m) on p-nitrophenyl-beta-d-xyloside was 0.047mm. the homogeneous xylanase (1,4-beta-d-xylan xylanohydrolase, ec 3.2.1.8) and beta-xylosidase showed differences in their mode of action towards xylooligosaccharides ...198618555300
localization of alkaline phosphatase activity at microbody membranes of neurospora crassa and aspergillus nidulans.hyphal cells of neurospora crassa and aspergillus nidulans, grown in sabouraud glucose broth or in a defined medium with xanthine or its catabolites as the nitrogen source, contained single membrane-bound organelles cytochemically identified as microbodies. modified gomori procedures at the ultrastructural level revealed putative alkaline phosphatase activity sites in thin sections of cells of both species of fungi. microbody membranes displayed electron opaque deposits (lead phosphate) which we ...19862948778
mitochondrial gene urfn of neurospora crassa codes for a long polypeptide with highly repetitive structure.the mitochondrial dna of neurospora crassa contains a long potential gene, designated urfn, which is located immediately downstream from the co1 gene. these two genes are encoded in different reading frames and overlap by 13 codons. urfn is 633 triplets long and terminates at a uag stop codon. its codon usage is atypical for n. crassa mitochondrial exons and introns, and resembles that of the long open reading frame (orf) of the mitochondrial plasmid present in n. crassa strain mauriceville. mul ...19862949084
sequence analysis and transformation by the catabolic 3-dehydroquinase (qute) gene from aspergillus nidulans.the induction of catabolic 3-dehydroquinase by quinic acid in aspergillus nidulans has been shown to involve transcriptional control and yields a single major 0.8 kb mrna. the nucleotide sequence of the catabolic 3-dehydroquinase qute gene has been determined and contains a single uninterrupted open reading frame of 462 bases encoding a 16,505 da protein of 153 residues. comparison with the corresponding qa2 gene of neurospora crassa reveals the absence of 75 nucleotides encoding 25 amino acids ...19862949740
purification and characterization of 3-dehydroquinase from escherichia coli.a procedure has been developed for the purification of 3-dehydroquinase from escherichia coli. homogeneous enzyme with specific activity 163 units/mg of protein was obtained in 19% overall yield. the subunit mr estimated from polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate was 29,000. the native mr, estimated by gel permeation chromatography on sephacryl s-200 (superfine) and on tsk g3000sw, was in the range 52,000-58,000, indicating that the enzyme is dimeric. the ...19862950851
neurospora crassa mitochondria contain two forms of a 4'-phosphopantetheine-modified protein.when neurospora crassa was labeled with [14c]pantothenic acid during growth, the mitochondrial fraction contained two bands of radioactivity of mr 19,000 and 22,000 by sodium dodecyl sulfate gel electrophoresis. the 19-kda band was converted to the 22-kda band by four treatments which are characteristic of the cleavage of a thioester bond: dithiothreitol and 2-mercaptoethanol at basic but not neutral ph, alkaline methanolysis, sodium borohydride in tetrahydrofuran, and hydroxylamine at neutral p ...19862937780
a study of the messenger rna encoding pyruvate kinase of neurospora crassa.in neurospora crassa, there is a single pyruvate kinase (pk) consisting of four identical subunits of approximately 60k daltons. northern and dot blot hybridization studies, using most of the yeast pyruvate kinase gene as a probe, suggest the presence of two distinct mrna species for pyruvate kinase, separable on the basis of the length of their polyadenylated tails, by oligo(dt)cellulose chromatography. these messages are present in polysomes, immuno-precipitated by anti-pk antibodies, indicati ...19862941081
a cloned tryptophan-synthesis gene from the ascomycete cochliobolus heterostrophus functions in escherichia coli, yeast and aspergillus nidulans.a gene (trp1) in the tryptophan biosynthetic pathway of the fungal plant pathogen cochliobolus heterostrophus was isolated by complementation of an escherichia coli trpf mutant which lacked phosphoribosylanthranilate isomerase (prai) activity. the cloned gene also complemented an e. coli trpc mutant lacking indoleglycerolphosphate synthase (igps) activity, a yeast trp1 mutant missing prai activity and an aspergillus nidulans trpc mutant. it functioned in e. coli and a. nidulans without apparent ...19862941339
the effect of papulacandin b on (1----3)-beta-d-glucan synthetases. a possible relationship between inhibition and enzyme conformation.the antibiotic, papulacandin b, inhibited growth or (1----3)-beta-d-glucan synthetase (or both) in the fungi saccharomyces cerevisiae, hansenula anomala, neurospora crassa, cryptococcus laurentii, schizophyllum commune and wangiella dermatitidis. no effect was observed on achlya ambisexualis. there was no apparent correlation between the inhibition of growth and that of the synthetase. with most of the fungal extracts, the inhibition of glucan synthetase by papulacandin b became less pronounced ...19862942248
biochemical comparison of the neurospora crassa wild type and the temperature-sensitive and leucine-auxotroph mutant leu-5. purification of the cytoplasmic and mitochondrial leucyl-trna synthetases and comparison of the enzymatic activities and the degradation patterns.the cytoplasmic leucyl-trna synthetases of neurospora crassa wild type (grown at 37 degrees c) and mutant (grown at 28 degrees c) were purified approximately 1770-fold and 1440-fold respectively. additional enzyme preparations were carried out with mutant cells grown for 24 h at 28 degrees c and transferred then to 37 degrees c for 10-70 h of growth. the mitochondrial leucyl-trna synthetase of the wild type was purified approximately 722-fold. the mitochondrial mutant enzyme was found only in tr ...19862942398
regulation of fungal cell wall growth: a guanine nucleotide-binding, proteinaceous component required for activity of (1----3)-beta-d-glucan synthase.by treatment with detergent and nacl, particulate (1----3)-beta-d-glucan synthase (ec 2.4.1.34) from hansenula anomala or neurospora crassa was dissociated into a "soluble fraction" and a "membrane fraction." each fraction alone was almost inactive, but enzymatic activity could be reconstituted by mixing the two fractions and adding gtp or one of its analogs. based on their lability to heat and to incubation with trypsin, the activity in both fractions is proteinaceous. the active component in t ...19862942941
compartmental and regulatory mechanisms in the arginine pathways of neurospora crassa and saccharomyces cerevisiae. 19862945985
isolation and partial nucleotide sequence of the laccase gene from neurospora crassa: amino acid sequence homology of the protein to human ceruloplasmin.the laccase (benzenediol:oxygen oxidoreductase, ec 1.10.3.2) gene from neurospora crassa was cloned and part of its nucleotide sequence corresponding to the carboxyl-terminal region of the protein has been determined. the gene was cloned by cdna synthesis with a laccase-specific synthetic deoxyundecanucleotide as primer and poly(a) rna isolated from cycloheximide-treated n. crassa cultures as template. based on the nucleotide sequence of the cdna obtained, a unique 21-mer was synthesized and use ...19862947240
labeling of individual amino acid residues in the membrane-embedded f0 part of the f1 f0 atp synthase from neurospora crassa. influence of oligomycin and dicyclohexylcarbodiimide.three f0 subunits and the f1 subunit beta of the atp synthase from neurospora crassa were labeled with the lipophilic photoactivatable reagent 3-(trifluoromethyl)-3-(m-[125i]iodophenyl)diazirine ([125i]tid). in the proteolipid subunit which was the most heavily labeled polypeptide labeling was confined to five residues at the nh2-terminus and five residues at the c-terminus of the protein. labeling occurred at similar positions compared with the homologous protein (subunit c) in the atp synthase ...19862869944
topological studies suggest that the pathway of the protons through f0 is provided by amino acid residues accessible from the lipid phase.the structure of the f0 part of atp synthases from e. coli and neurospora crassa was analyzed by hydrophobic surface labeling with [125i]tid. in the e. coli f0 all three subunits were freely accessible to the reagent, suggesting that these subunits are independently integrated in the membrane. labeled amino acid residues were identified by edman degradation of the dicyclohexylcarbodiimide binding (dccd) proteins from e. coli and neurospora crassa. the very similar patterns obtained with the two ...19862874840
primary structure of the neurospora plasma membrane h+-atpase deduced from the gene sequence. homology to na+/k+-, ca2+-, and k+-atpase.the gene for the neurospora crassa plasma membrane h+-atpase has been cloned and sequenced. the gene encodes for a protein of 920 amino acids with a molecular weight of 100,002. the coding region is interrupted by four introns: three near the amino terminus and one near the carboxyl terminus. the deduced amino acid sequence of the n. crassa plasma membrane h+-atpase exhibits 75% homology to the amino acid sequence of the saccharomyces cerevisiae plasma membrane h+-atpase. also, an amino acid com ...19862876992
the atp synthase subunit 9 gene of aspergillus nidulans: sequence and transcription.we have determined the nucleotide sequence of the aspergillus nidulans nuclear gene olic31, which encodes subunit 9 of mitochondrial atp synthase. the open reading frame contains no introns and specifies a predicted protein of 143 amino acids comprising a pre-sequence of 62 residues and a mature protein of 81 residues. the amino acid homology with the equivalent neurospora crassa protein is 50% for the pre-sequence and 80% for the mature protein. a comparison with this and other imported mitocho ...19862880279
yeast mitochondrial atpase subunit 8, normally a mitochondrial gene product, expressed in vitro and imported back into the organelle.subunit 8 of yeast mitochondrial f1f0-atpase is a proteolipid made on mitochondrial ribosomes and inserted directly into the inner membrane for assembly with the other f0 membrane-sector components. we have investigated the possibility of expressing this extremely hydrophobic, mitochondrially encoded protein outside the organelle and directing its import back into mitochondria using a suitable n-terminal targeting presequence. this report describes the successful import in vitro of atpase subuni ...19862881782
molybdopterin cofactor from methanobacterium formicicum formate dehydrogenase.the molybdopterin cofactor from the formate dehydrogenase of methanobacterium formicicum was studied. the cofactor was released by guanidine denaturation of homogeneous enzyme, which also released greater than 80% of the molybdenum present in the enzyme. the anoxically isolated cofactor was nonfluorescent, but after exposure to air it fluoresced with spectra similar to those of described molybdopterin cofactors. aerobic release from acid-denatured formate dehydrogenase in the presence of i2 and ...19863700335
purification and properties of nitrate reductase from mitsuokella multiacidus.nitrate reductase of mitsuokella multiacidus (formerly bacteroides multiacidus) was solublized from the membrane fraction with 1% sodium deoxycholate and purified 40-fold by immunoaffinity chromatography on the antibody-affi-gel 10 column. the preparation showed a major band (86% of total protein) with enzyme activity and a minor band on polyacrylamide gel after disc electrophoresis in the presence of 0.1% triton x-100. sodium dodecyl sulfate-polyacrylamide gel electrophoresis gave a major band, ...19863711052
the simple repeat poly(dt-dg).poly(dc-da) common to eukaryotes is absent from eubacteria and archaebacteria and rare in protozoans.genomic dna from a wide variety of prokaryotic and eukaryotic organisms has been assayed for the simple repeat sequence poly(dt-dg).poly(dc-da) by southern blotting and dna slot blot hybridizations. consistent with findings of others, we have found the simple alternating sequence to be present in multiple copies in all organisms in the animal kingdom (e.g., mammals, reptiles, amphibians, fish, crustaceans, insects, jellyfish, nematodes). the tg element was also found in lower eukaryotes (sacchar ...19863127653
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
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
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
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
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
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
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
behaviour of recombinant plasmids in aspergillus nidulans: structure and stability.a pyrg- aspergillus strain was transformed with plasmid pdjb-1, derived from pbr325 by insertion of the neurospora crassa pyr4 gene (orotidine 5'-phosphate carboxylase), giving mitotically unstable transformants. aspergillus dna which acted as an "autonomously replicating sequence" (ars) in yeast was inserted into pdjb-1 and the resulting construct, pdjb12.1, gave mitotically stable transformants when introduced into aspergillus. transformants obtained with pdjb-1 and pdjb12.1 gave few pyr- prog ...19863329034
molybdenum cofactor: a compound in the in vitro activation of both nitrate reductase and trimethylamine-n-oxide reductase activities in escherichia coli k12.nitrate reductase (nitrite: (acceptor) oxidoreductase, ec 1.7.99.4) and trimethylamine n-oxide reductase (nadh : trimethylamine-n-oxide oxidoreductase, ec 1.6.6.9) activities were reconstituted by incubation of the association factor fa (the putative product of the chlb gene) with the soluble extract of the chlb mutant grown anaerobically in the presence of trimethylamine n-oxide. when soluble extracts of the chlb mutant grown on 10 mm sodium tungstate, a molybdenum competitor, were used in comp ...19863524687
activation in vitro of respiratory nitrate reductase of escherichia coli k12 grown in the presence of tungstate. involvement of molybdenum cofactor.the chlorate-resistant (chlr) mutants are pleiotropically defective in molybdoenzyme activity. the inactive derivative of the molybdoenzyme, respiratory nitrate reductase, present in the cell-free extract of a chlb mutant, can be activated by the addition of protein fa, the probable active product of the chlb locus. protein fa addition, however, cannot bring about the activation if 10 mm sodium tungstate is included in the culture medium for the chlb strain. the inclusion of a heat-treated prepa ...19863525161
further characterization of trimethylamine n-oxide reductase from escherichia coli, a molybdoprotein.escherichia coli trimethylamine n-oxide (tmao) reductase i, the major enzyme among inducible tmao reductases, was purified to homogeneity by an improved method including heat treatment, ammonium sulfate precipitation, and chromatographies on bio-gel a-1.5m, deae-cellulose, and reactive blue-agarose. the molecular weight was estimated by gel filtration to be approximately 200,000. a single subunit peptide with a molecular weight of 95,000 was found by sodium dodecyl sulfate-polyacrylamide gel ele ...19863528139
simultaneous purification of three mitochondrial enzymes. acetylglutamate kinase, acetylglutamyl-phosphate reductase and carbamoyl-phosphate synthetase from neurospora crassa.the early enzymes of arginine biosynthesis in neurospora crassa are localized in the mitochondrion and catalyze the conversion of glutamate to citrulline. the final conversion of citrulline to arginine occurs via two enzymatic steps in the cytoplasm. we have devised a method for the isolation and purification of three of the mitochondrial arginine biosynthetic enzymes from a single extract. acetylglutamate kinase and acetylglutamyl-phosphate reductase (both products of the complex arg-6 locus) w ...19862420793
h+/atp stoichiometry of proton pumps from neurospora crassa and escherichia coli.a kinetic method has been used to measure the apparent stoichiometry of h+ ions translocated per atp split by membrane-bound [h+]-atpases. in this method, membrane vesicles are suspended in well-buffered medium, atp is added, and a fluorescent probe of delta ph (acridine orange) is used to detect the formation of a steady-state ph gradient. at the steady state, it is assumed that proton pumping in one direction is exactly balanced by the leak of protons in the opposite direction. the pump is the ...19862425739
immunological cross-reactivity of fungal and yeast plasma membrane h+-atpase.the plasma membrane h+-atpases from fungi and yeasts have similar catalytic and molecular properties. a structural comparison has been performed using immunoblot analysis with polyclonal antibodies directed toward the 102 kda polypeptide of the plasma membrane h+-atpase from neurospora crassa. a strong cross-reactivity is observed between the fungal h+-atpase and the enzyme from the yeasts saccharomyces cerevisiae and schizosaccharomyces pombe. structural homologies are indicated also by the ana ...19862428661
inversions and recombinations in mitochondrial dna of the (sg-1) cytoplasmic mutant in two neurospora species.the mitochondrial dnas of [sg-1] cytoplasmically-mutant and wild-type strains of neurospora crassa and neurospora sitophila were examined by comparative restriction endonuclease analyses. the mtdna of n. sitophila wild type of whitehouse differs from type ii mtdna of n. crassa by insertions of 3.3 kb in ecori-9, and 1.2 kb in ecori-3, and a deletion of 1.1 kb in ecori-5. these dna heteromorphisms provided convenient markers for tracing n. crassa [sg-1] mtdna during and after its transfer into n. ...19862832078
isolation and identification of the aspergillus nidulans gdha gene encoding nadp-linked glutamate dehydrogenase.the neurospora crassa am gene was used as a heterologous probe to identify clones from two independently constructed aspergillus nidulans gene libraries. these clones have a common hindiii 1.85 kb fragment. this a. nidulans nucleotide stretch hybridises to a n. crassa 2.7 kb bamhi fragment of wild type dna but not to a co-migrating fragment from the dna of the n. crassa am132 deletion mutant. one a. nidulans clone was shown to complement the n. crasse am132 deletion strain. the n. crassa transfo ...19862834076
primary structure of the neurospora crassa small subunit ribosomal rna coding region. 19862948156
ascus development in two temperature-sensitive four-spore mutants of neurospora crassa.two nonallelic four-spore mutants are known in which ascospore walls enclose the four immediate products of meiosis rather than the normal eight products of a postmeiotic mitosis. expression depends on temperature. the four-spore phenotype is expressed when the developing asci are subjected either to high temperatures (25-30 degrees c) for fsp-1 or to low temperatures (15-20 degrees c) for fsp-2. heterozygous fsp-1 x fsp-1+ crosses make eight-spored asci at 15-20 degrees c but produce many four- ...19862950989
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