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isolation of nit-4, the minor nitrogen regulatory gene which mediates nitrate induction in neurospora crassa.expression of nitrate reductase in neurospora crassa requires the positive action of nit-4, a pathway-specific regulatory gene, which mediates nitrate induction. we report the molecular cloning of the nit-4 gene and present results which suggest that the nit-4 gene is constitutively expressed to yield a low-abundance 2.2-kilobase transcript. these results indicate that the nit-2 major control gene and the nit-4 pathway-specific control gene independently regulate the expression of the nitrate as ...19892567729
effect of the uvs-2 allele of neurospora crassa on the mutagenic potency of two n-hydroxylaminopurines and 2-aminopurine in the ad-3 forward-mutation test.the mutagenic potencies of 3 purine analogs were determined in the ad-3 forward-mutation test in growing cultures of heterokaryon 59 (h-59), a nucleotide excision repair-deficient (uvs-2/uvs-2) 2-component heterokaryon of neurospora crassa. two n-hydroxylaminopurines, 2-amino-6-n-hydroxylaminopurine (aha) and 6-n-hydroxylaminopurine (hap), were potent and strong mutagens, respectively, whereas 2-aminopurine (ap) was a moderate mutagen. dose-response curves showed that aha and hap were about equa ...19892526296
the response time of transcription and translation of the leu-2 gene of neurospora to its inducer, alpha-isopropylmalate, approaches the permissible minimum.the rate of transcription and translation of the leu-2 gene of neurospora crassa was measured after induction by alpha-isopropylmalate. little message of enzyme was found before inducer addition but transcription in the lower eukaryote was found well underway within five minutes after inducer addition, followed in a minute or two by the appearance of functional enzyme. the timing was close to the limit set by rna synthesis and ribosome procession. as a consequence, it seems unlikely that travers ...19892525903
use of transformation to make targeted sequence alterations at the am (gdh) locus of neurospora.specific in vitro-generated insertion, replacement, and deletion mutations have been integrated near the chromosomal locus of am (nadp-specific glutamate dehydrogenase) of neurospora crassa. two approaches have been successful. one approach used am+-containing vectors capable of integrating at any site in the genome. this technique was used to introduce a specific 700 bp insertion near the am locus and to replace chromosomal sequences near am with plasmid dna. efficiency was low, however, and ma ...19892549376
early response and induced tolerance to cycloheximide in neurospora crassa.incubation of neurospora crassa mycelia with low doses of cycloheximide induces the expression of several genes. after 6 h in the presence of cycloheximide, mycelia become tolerant to further additions of the drug and the rate of protein synthesis exhibits a lower sensitivity to it. the polypeptide pattern is indicative of a stress situation.19892528413
effects of heat shock on the induction of mutations by chemical mutagens in neurospora crassa.preheating of neurospora conidia increased their susceptibility to mutation induction by chemical mutagens. optimal conditions of heat shock for enhanced mutagenesis were determined in 2.5 x 10(7) conidia/ml 0.067 m kh2po4-na2hpo4 (ph 7.0) buffer to be treatment at 43 degrees c for 60 min. when protein synthesis during heat stock was eliminated by cycloheximide or by use of the temperature-sensitive mutation psi-1, induction of thermotolerance was inhibited while induction of the enhanced state ...19892525670
uptake, intracellular binding, and excretion of polyamines during growth of neurospora crassa.in neurospora crassa mycelia, the amounts of the main polyamines, putrescine and spermidine, are approximately 0.8 and 18 nmol/mg, dry weight. we wished to know what determines these pool sizes. in the growth medium, externally added polyamines enter cells largely by a nonsaturable, diffusional system. in a mutant unable to polyamines, internal and external spermidine appear to equilibrate across the cell membrane during growth. however, this was true only after an intracellular "sink," with a c ...19892524999
independent transfer of mitochondrial plasmids in neurospora crassa.in the ascomycete fungus neurospora, the distribution of homologous mitochondrial plasmid dnas in different species and among mitochondrial types of n. crassa suggests that these molecules have moved between lineages of clonally propagated mtdna. here we report direct evidence for independent inheritance of mitochondrial plasmids by sexual reproduction which may help explain the distribution of these molecules among mitochondrial lineages.19892524667
fusion of the mitochondrial outer membrane: use in forming large, two-dimensional crystals of the voltage-dependent, anion-selective channel protein.phospholipase a2 induces crystallization of the channel protein, vdac (also called mitochondrial porin), in the outer membrane of neurospora crassa mitochondria. the channel crystals formed in native membranes typically contain a few hundred unit cells. to increase the size of these membrane crystals for low-contrast electron microscopic imaging and diffraction studies, fusion of the isolated mitochondrial outer membranes was attempted before and after phospholipase treatment. successful fusion ...19892470408
luminescence emission from neurospora copper metallothionein. time-resolved studies.the luminescence lifetime of cu-metallothionein from the fungus neurospora crassa has been studied by the frequency-domain emission technique. lifetimes of 10.3 and 3.4 microseconds have been found for the protein in the absence and in the presence of oxygen respectively. binding of hg(ii) results in a quenching of the luminescence correlated to the shortening of lifetime to 0.3-0.4 microsecond. no quenching by oxygen is found for the hg(ii)-cu-metallothionein adduct. by analogy to model compoun ...19892528343
dna sequence, organization and regulation of the qa gene cluster of neurospora crassa.in neurospora, five structural and two regulatory genes mediate the initial events in quinate/shikimate metabolism as a carbon source. these genes are clustered in an 18 x 10(3) base-pair region as a contiguous array. the qa genes are induced by quinic acid and are coordinately controlled at the transcriptional level by the positive and negative regulators, qa-1f and qa-1s, respectively. the dna sequence of the entire qa gene cluster has been determined and transcripts for each gene have been ma ...19892525625
identification of an arginine carrier in the vacuolar membrane of neurospora crassa.a number of arginine derivatives were tested for their ability to inhibit arginine uptake into vacuolar membrane vesicles of neurospora crassa. the guanido side chain and l-configuration were found to be important for recognition by the arginine carrier. based upon the specificity of recognition, a reactive arginine derivative (n alpha-p-nitrobenzyloxycarbonyl arginyl diazomethane) was synthesized which has an intact guanido side chain and a diazo group at the carboxyl end. the latter decomposes ...19892523392
identification and electron microscopic analysis of a chaperonin oligomer from neurospora crassa mitochondria.a 7-fold symmetric particle has been identified in neurospora crassa which is most probably the mitochondrial chaperonin. the particle, about 12 nm in diameter, appears in preparations of cytochrome reductase, and is shown to contain a 60 kd protein which cross-reacts with anti-groel antibodies. results of stem mass measurement suggest that the particle is composed of 14 subunits. a preliminary interpretation of the structure of the particle based on electron microscopy is given. its quaternary ...19892569968
premeiotic disruption of duplicated and triplicated copies of the neurospora crassa am (glutamate dehydrogenase) gene.premeiotic inactivation of duplicated sequences (the rip phenomenon of selker et al.) was studied by tetrad analysis using ectopic copies of am+ (coding for nadp-specific glutamate dehydrogenase) and a missense allele am3, coding for a distinctive form of the enzyme, at the normal locus. in duplication crosses either both gene copies were inactivated or neither. two inactivated am3 derivatives were shown to have undergone methylation and numerous base-pair changes, reflected in losses and gains ...19892529044
bioelectrorheological model of the cell. 2. analysis of creep and its experimental verification.the electrorheological model of the cell proposed in part 1 of this work was used to analyze changes in time of the shape of a cell acted on by a constant-amplitude external alternating electric field, with lossiness of the media taken into account. shear stress in the cell membrane was determined. this model was then subjected to preliminary experimental verification using neurospora crassa (slime) spheroplasts subjected to an external alternating electric field of constant frequency (3 mhz) an ...19892533955
proton nmr studies of a metallothionein from neurospora crassa: sequence-specific assignments by noe measurements in the rotating frame.sequential 1h nmr assignments of a metallothionein from neurospora crassa have been accomplished by the combined use of cosy, 2qf-cosy, hohaha, and rotating-frame noe experiments. all potentially observable resonances were assigned except for the epsilon-nh3 group of the c-terminal lysine. 1h noes, when observed in the laboratory frame and at 500-mhz spectrometer frequency, were negligible in this protein due to the inherent rotational correlation time of the molecule. this difficulty was circum ...19892525920
two kinds of "recombination nodules" in neurospora crassa.two morphological types of recombination nodules, termed early and late, are recognized in neurospora crassa. eighty nuclei at different substages were used to determine numbers of nodules per nucleus, distribution of nodules along the nucleolus-organizing chromosome, and distribution of nodules among the two largest chromosomes. early nodules appear at the synaptonemal complex at early zygotene and increase in number during zygotene until a dramatic reduction occurs at zygotene-pachytene transi ...19892526043
a morphological and genetic analysis of conidiophore development in neurospora crassa.the filamentous fungus neurospora crassa responds to nutrient deprivation and dessication by producing asexual spores, or conidia. these conidia are derived from differentiated aerial structures called conidiophores. the process of conidiation was analyzed in wild-type and morphological mutants using scanning electron microscopy (sem) and specific fluorescent probes. the first discernible morphological step of conidiation is the transition from growth by hyphal tip elongation to growth by repeat ...19892524423
change in chromosome number associated with a double deletion in the neurospora crassa mitochondrial chromosome.the mitochondrial genome of neurospora is usually found in a single covalently closed circular 62-kbp dna molecule. we report here that the mitochondrial genome of a phenotypic revertant of a stopper mutant (stp-ruv) is contained primarily in two separate, nonoverlapping, autonomously replicating circular chromosomes. the circles, one about 21 kbp and the other somewhat less than 36 kbp are derived from the most frequent classes of recombinant chromosomes (21 and 41 kbp) in the chromosomal popul ...19892524420
characterization of nitrate reductase deficient mutants of chlorella sorokiniana.after x-ray irradiation, 13 mutants of chlorella sorokiniana incapable of using no(3) (-) as n source were isolated using a pinpoint method. using immunoprecipitation and western blot assays, no nitrate reductase was found in five strains while in eight mutants the enzyme was detected. the latter strains contained different patterns of nitrate reductase partial reactions. all isolates were of the nia-type as indicated by the inducibility of purine hydroxylase i and by complementation of nitrate ...198916666622
isolation of a transposable element from neurospora crassa.a neurospora crassa strain from adiopodoumé, ivory coast, contains multiple copies of a transposable element, tad. the element was detected as a 7-kilobase insertion in two independently isolated spontaneous forward mutants of the am (glutamate dehydrogenase) gene. laboratory strains do not contain tad. all progeny from crosses of the adiopodoumé strain to laboratory strains contain multiple copies. when the element was inserted in am, target sequences of 14 and 17 base pairs were duplicated in ...19892538822
isolation and characterization of cadmium-resistant mutants of neurospora crassa.this study identified and characterized four cadmium-resistant mutants of neurospora crassa. one of these mutants maps to linkage group ii and the other three map to linkage group vii, whereas a naturally occurring resistant trait in a strain from japan resides at a distinct but unmapped locus. transport of cadmium into neurospora cells occurs by more than a single uptake system and involves both energy-dependent and -independent components. the resistant mutants transport cadmium in the same ma ...19892525066
immunological identification of the alternative oxidase of neurospora crassa mitochondria.neurospora crassa mitochondria use a branched electron transport system in which one branch is a conventional cytochrome system and the other is an alternative cyanide-resistant, hydroxamic acid-sensitive oxidase that is induced when the cytochrome system is impaired. we used a monoclonal antibody to the alternative oxidase of the higher plant sauromatum guttatum to identify a similar set of related polypeptides (mr, 36,500 and 37,000) that was associated with the alternative oxidase activity of ...19892524649
a small isoform of nadh:ubiquinone oxidoreductase (complex i) without mitochondrially encoded subunits is made in chloramphenicol-treated neurospora crassa.in mitochondria of neurospora crassa grown in the presence of chloramphenicol a small form of nadh:ubiquinone reductase is made in place of the normal electron-transfer-complex i. this smaller enzyme has a molecular mass of approximately 350 kda and consists of (at least) 13 different subunits which are all synthesized in the cytoplasm. the complex i which is normally found in neurospora has a molecular mass of approximately 700 kda and consists of around 30 different subunits, of which at least ...19892523306
13n isotope studies of glutamine assimilation pathways in neurospora crassa.l-[amide-13n]glutamine in neurospora crassa is metabolized to [13n]glutamate by glutamate synthase and to [13n]ammonium by the glutamine transaminase-omega-amidase pathway. the [13n]ammonium released is assimilated by glutamate dehydrogenase and glutamine synthetase, confirming the operation of a glutamine cycle. most of the nitrogen is retained during cycling between glutamate and glutamine.19892522094
in vivo control of gluconeogenesis in wild-type neurospora crassa and in the adenylate cyclase-deficient cr-1 (crisp) mutant.the rate of cycloheximide-resistant incorporation of carbon from [14c]alanine and [14c]acetate into polysaccharidic material was used to study gluconeogenic activity in wild-type neurospora crassa and in the adenylate cyclase-deficient cr-1 (crisp-1) mutant. the wild-type efficiently utilized alanine and acetate as gluconeogenic substrates, whereas the mutant used acetate efficiently but was unable to use alanine. cycloheximide-resistant 14c-incorporating activity was sensitive to carbon catabol ...19892522093
x-ray-induced specific-locus mutations in the ad-3 region of two-component heterokaryons of neurospora crassa. ii. more extensive genetic tests reveal an unexpectedly high frequency of multiple-locus mutations.more extensive genetic tests have been performed on a series of 832 x-ray-induced specific-locus mutations in the ad-3 region of a 2-component heterokaryon (h-12) of neurospora crassa, reported earlier (webber and de serres 1965). using new tester strains and techniques for performing large-scale genetic tests (heterokaryon, dikaryon and trikaryon) to characterize ad-3 mutants induced in 2-component heterokaryons, new data have been obtained on this sample of x-ray-induced ad-3 mutants. these ne ...19892521371
neurospora crassa alpha-ketoglutarate dehydrogenase complex: description, resolution of components and catalytic properties.a method is proposed for the purification of the neurospora crassa alpha-ketoglutarate dehydrogenase complex, and the main points for preserving its activity, which seems to be particularly fragile in fungus, are discussed. resolution of the constitutive enzymes was attempted and permitted the identification of the three protein bands resolved on sds-polyacrylamide gel electrophoresis as e3, e1 and e2 with respective mr values of 54,000, 53,000 and 49,000. catalytic properties of the purified co ...19892521564
sedimentation properties of chitosomal chitin synthetase from the wild-type strain and the 'slime' variant of neurospora crassa.marked differences in the pattern of sedimentation of cellular structures were observed after isopycnic centrifugation of crude cell-free preparations from the neurospora crassa wall-less 'slime' variant and mycelial wild-type strain. kinetic studies of particle sedimentation showed that the various types of subcellular components, as revealed by turbidity, uv absorption, polypeptide patterns, and chitin synthetase activity determinations, sediment independently of one another. an important feat ...19892521563
nad-specific glutamate dehydrogenase of neurospora crassa. cdna cloning and gene expression during derepression.the catabolic nad-specific glutamate dehydrogenase of neurospora crassa is one of the many enzymes regulated by carbon catabolite repression. to achieve an understanding of its regulation, cdna and genomic clones were isolated. total poly(a+) rna from derepressed cells was used for the construction of a cdna library in the expression vector, lambda gt11. by screening this library with a polyclonal antiserum against nad-specific glutamate dehydrogenase, a positive clone with a 0.9-kilobase insert ...19892521336
genes expressed during conidiation in neurospora crassa: characterization of con-8.the filamentous fungus neurospora crassa, by a series of defined changes, differentiates from a mycelium composed of branching hyphae to form dormant spores, called conidia. several genes of unknown function (con genes) that are preferentially expressed during this period have been cloned. transcription of these genes has been examined in conidiation-defective mutants, and the results obtained revealed that con-6, con-8, con-10, con-11 and con-13 are most likely to play a unique role during coni ...19892521382
the biological clock of neurospora in a microgravity environment.the circadian rhythm of conidiation in neurospora crassa is thought to be an endogenously derived circadian oscillation; however, several investigators have suggested that circadian rhythms may, instead, be driven by some geophysical time cue(s). an experiment was conducted on space shuttle flight sts-9 in order to test this hypothesis; during the first 7-8 cycles in space, there were several minor alterations observed in the conidiation rhythm, including an increase in the period of the oscilla ...198911537340
isolation of everted plasma membrane vesicles from neurospora crassa and measurement of transport function. 19892561173
phase determination of the circadian rhythm of conidiation in heterocaryons between two out-of-phase mycelia in neurospora crassa.neurospora grows vegetatively as a syncytium in which multiple nuclei exist within a connected cytoplasm. because of the ability of separate and distinct mycelia to fuse, the possibility exists of generating heterocaryotic cultures in which the nuclei and cytoplasms of two different strains are comingled into the same syncytium. we have used such heterocaryons, in which the component parts differed with respect to their circadian clock phase, to examine whether or not clock-dominant phases exist ...19892535268
[antifungal activity of 5-benzilidene pyrrolone and furanone derivatives].the antifungal activity against neurospora crassa of some 5-benzilidene pyrrolone and furanone derivatives was realised. relations between the structure and this biological activity are established with fujita-ban and hansch methods. the preponderant part of lipophilicity, resonance effect and e or z configurations have been showed.19892535109
iron limitation and its effect on membrane proteins and siderophore transport in neurospora crassa.cells of the fungus neurospora crassa were grown under iron-deficient and iron-sufficient conditions and their plasma membrane proteins were compared. three strains were studied: n. crassa 74a (wild type), a siderophore-free mutant n. crassa (arg-5 ota aga) as well as a 'slime' variant of n. crassa which lacks a cell wall. plasma membranes were purified, solubilized and analyzed by one-dimensional sds/polyacrylamide gel electrophoresis yielding approximately 50 distinct protein bands with molecu ...19892534965
acid phosphatase (ec 3.1.3.2) synthesis by phosphorus regulatory mutant strains of neurospora crassa.1. even though altered forms of acid phosphatase ii were synthesized by the mutant strains nuc-1a and nuc-2a of n. crassa, their synthesis was independent of exogenous phosphate concentrations. 2. synthesis of acid phosphatase i by nuc-2a was also insensitive to exogenous phosphate concentrations. when nuc-1a was grown on a low-phosphate medium, it also produced a heat-labile acid phosphatase in addition to a i-like acid phosphatase. i-like acid phosphatase was not detected in the mycelium of th ...19892531620
duplication of the trna(mmet) and trna(cys) genes and of fragments of a gene encoding a subunit of the nadh dehydrogenase complex in neurospora grassa mitochondrial dna.neurospora crassa mitochondrial dna (mtdna) contains duplications of the trna(mmet) gene upstream of a gene (nd2) encoding a subunit of the nadh dehydrogenase complex and of the trna(cys) gene which is found downstream of the apocytochrome b gene. both duplicated genes are located upstream of the small rrna gene. the duplications are extended to flanking sequences. in the case of the trna(mmet) duplication, two fragments of the nd2 gene are also duplicated. these two fragments, which are not con ...19892525962
deoxyglucose-resistant mutants of neurospora crassa: isolation, mapping, and biochemical characterization.neurospora crassa mutants resistant to 2-deoxyglucose have been isolated, and their mutations have been mapped to four genetic loci. the mutants have the following characteristics: (i) they are resistant to sorbose as well as to 2-deoxyglucose; (ii) they are partially or completely constitutive for glucose transport system ii, glucamylase, and invertase, which are usually repressed during growth on glucose; and (iii) they synthesize an invertase with abnormal thermostability and immunological pr ...19892521617
sensitivity to bleomycin and hydrogen peroxide of dna repair-defective mutants in neurospora crassa.mutations were induced in neurospora which cause increased sensitivity to mms (methyl methane-sulfonate) and other mutagens. genetic analysis of such mus demonstrated that some of them defined new dna repair genes (mus-21, and mus-27 to mus-30), while others represented new alleles in previously known genes. to characterize them further, and especially to identify rec- types which have not yet been found in this species, many mms-sensitive strains were tested for cross-sensitivities to bleomycin ...19892463486
nucleotide sequence of a neurospora crassa ribosomal protein gene. 19902147995
nucleotide sequence of the exons of the large subunit rrna of neurospora crassa mitochondria. 19901701879
genes expressed during conidiation in neurospora crassa: molecular characterization of con-13.asexual development in neurospora crassa proceeds through a series of discrete morphological stages that culminate in the production of dormant spores called conidia. changes in the pattern of gene expression parallel the morphological transformations associated with conidiation. as a prerequisite to the analysis of developmental gene expression in n. crassa, several genes of unknown function that are preferentially expressed during conidiation were isolated [berlin and yanofsky, mol. cell. biol ...19902148538
glutamine metabolism and cycling in neurospora crassa.[this corrects the article on p. toc in vol. 54.].199016350247
5-benzylidene pyrrolones. iv--synthesis and antifungal activity of some 5-benzylidene derivatives of 1,2-dimethyl-3-carbethoxy-pyrrol-4-one.in order to investigate the antifungal activity of pyrrolones, the synthesis of derivatives of 5-benzylidene-1,2,-dimethyl-3-carbethoxy-pyrrol-4-one was achieved by a one-step reaction. the condensation reaction was applied to the above-mentioned heterocycle and seven substituted benzaldehydes were obtained providing the entitled compounds, of which six are original. they have preferential e configuration. antifugal data against neurospora crassa in comparison with ketoconazole showed that many ...19902151022
generation of new functional mutant alleles by premeiotic disruption of the neurospora crassa am gene.in the further analysis of a cross in which the mis-sense allele, am3, of the neurospora crassa am (glutamate dehydrogenase) gene was present in one parent together with two ectopic wild-type gene copies, one ascus was identified in which the two ectopic copies had been inactivated by the rip process whereas the am3 allele continued to produce its characteristic enzyme variety in active, but heat-sensitive, form. the am3 allele had also acquired a new hindiii restriction site. it had no detectab ...19902150348
cytosolic calcium homeostasis in fungi: roles of plasma membrane transport and intracellular sequestration of calcium.cytosolic free calcium ([ca2+]c) has been measured in the mycelial fungus neurospora crassa with ca2(+)-selective microelectrodes. the mean value of [ca2+]c is 92 +/- 15 nm and it is insensitive to external ph values between 5.8 and 8.4. simultaneous measurement of membrane potential enables the electrochemical potential difference for ca2+ across the plasma membrane to be estimated as about -60 kj.mol-1-a value that cannot be sustained either by a simple ca2(+)-atpase, or, in alkaline condition ...19902147513
regulation of branched-chain amino acid biosynthesis in neurospora crassa: cloning and characterization of the leu-1 and ilv-3 genes.the genes coding for the branched-chain amino acid biosynthetic enzymes comprise an integrated regulatory system. the expression of the several structural genes coding for enzymes of the isoleucine-valine and leucine pathways is controlled in parallel by the positive-acting regulatory gene, leu-3. the leu-1 and ilv-3 genes, coding for beta-isopropyl-malate dehydrogenase and aceto-hydroxyacid synthase, respectively, were cloned from a cosmid library. restriction fragment length polymorphism analy ...19901980003
studies on the sedimentation behavior of the neurospora crassa plasma membrane h(+)-atpase synthesized in vitro and integrated into homologous microsomal membranes.rna transcripts that encoded the neurospora crassa plasma membrane h(+)-atpase (pma+), a polytopic integral membrane protein, and the pma+344, a truncated pma+ with the amino terminal 344 amino acids, were translated in a n. crassa in vitro system. the microsomal membranes integrated products were insensitive to extraction by na2co3 (ph 11.5). the velocity sedimentation behavior of the in vitro synthesized pma+ were examined under various conditions. the pma+ migrated on linear sucrose gradients ...19902148269
expression of tyrosinase in vegetative cultures of neurospora crassa transformed with a metallothionein promoter/protyrosinase fusion gene.wild-type neurospora crassa, strain singapore, was transformed with a n. crassa metallothionein promoter/protyrosinase fusion gene. transformants produced tyrosinase during vegetative growth, as determined by western analyses and activity assays. this is in sharp contrast to wild-type strains, where this enzyme is only expressed in situations of starvation or sexual differentiation. complete integration of a 400 bp metallothionein promoter-fragment leads to constitutive expression of protyrosina ...19902147580
characterization of a novel plasmid-like element in neurospora crassa derived mostly from the mitochondrial dna.we have identified a plasmid-like element within mitochondria of neurospora crassa strain stp-b1. it is derived from the ecori-4 and ecori-6 regions of the mitochondrial dna, and an additional 124 bp dna segment of unknown origin. the plasmid dna consists of an oligomeric series of circular molecules of monomer length 2.2 kbp. the abundance of the plasmid suggests its autonomous replication and the presence of an efficient origin of replication. an unusually large number of palindromes capable o ...19902145549
molecular cloning of the l-amino-acid oxidase gene from neurospora crassa.the addition of d-phenylalanine to starved cultures of neurospora crassa leads to de novo synthesis of l-amino-acid oxidase. poly(a) rna from d-phenylalanine-treated mycelium was therefore used to generate a cdna library which was subsequently screened by hybrid-selected translation. a positive l-amino-acid oxidase clone served as a probe to isolate the complete gene from a genomic library of n. crassa. the nucleotide sequence obtained revealed an open reading frame coding for a protein of 695 a ...19902145270
tad, a line-like transposable element of neurospora, can transpose between nuclei in heterokaryons.the tad transposon of neurospora crassa appears to be a line-like element with very restricted distribution within the genus neurospora. when forced heterokaryons were constructed between strains which did and did not contain tad, the nuclei of the naive nuclear type rapidly acquired tad elements. the elements acquired by naive nuclei are active, since they can pass tad to other naive nuclei in subsequent heterokaryons. when heterokaryons are passaged by serial transfer, the load of acquired tad ...19902174012
heat shock response in neurospora crassa: purification and some properties of hsp 80.the heat shock response of neurospora crassa was investigated. a 80-kilodalton heat shock protein (hsp 80) was purified to near homogeneity from heat-shocked mycelial extracts employing ammonium sulphate fractionation, gel filtration, and ion-exchange and affinity chromatography. it was observed to migrate as a single band on one-dimensional sodium dodecyl sulphate--polyacrylamide gels, with a molecular mass of approximately 83 kilodaltons (kda). on two-dimensional gels it resolved into four pol ...19902148482
cpc-2, a new locus involved in general control of amino acid synthetic enzymes in neurospora crassa.in neurospora crassa starvation for single amino acids leads to derepression of enzymes in many amino acid synthetic pathways. regulation occurs at the level of transcription via "general amino acid (or cross-pathway) control". in this paper a new regulatory gene, cpc-2, is described that specifies a positive, trans-acting effector involved in this control. this gene, located on linkage group vii, was identified by a recessive mutation, u142, which results in sensitivity for two amino acid analo ...19902147403
the nucleotide excision repair epistasis group in neurospora crassa.dna repair mutants in eucaryotes are normally assigned to three epistasis groups. each epistasis group represents a "pathway" for dna repair. the pathways are commonly designated (1) nucleotide excision repair, (2) recombination repair and (3) mutagenic repair. an excision repair epistasis group has been established in neurospora and the mutants assigned to this group should be limited in their ability to excise pyrimidine dimers and other bulky lesions from dna. using a pyrimidine dimer-specifi ...19902147402
expansion and contraction of the nucleolus organizer region of neurospora: changes originate in both proximal and distal segments.previously we have shown that the nucleolus organizer region (nor) of neurospora crassa changes size frequently during the premeiotic portion of the sexual phase. here, we have investigated whether these changes in size originate only in specific regions of the nor, or are distributed throughout the nor. in two special strains of neurospora, the nor was divided into proximal and distal segments. in the first, the nor was divided by a translocation breakpoint and, in the second, the nor was divid ...19902147160
x-ray-induced specific-locus mutations in the ad-3 region of two-component heterokaryons of neurospora crassa. vii. genetic lesions resulting in gene/point mutations at the ad-3b locus have different dose-response relationships.genetic characterization of x-ray-induced ad-3 mutants, induced in a two-component heterokaryon (h-12) of neurospora crassa, has been performed to determine genotype, patterns of allelic complementation, and leakiness, and to distinguish gene/point mutations from multilocus deletions and multiple locus mutations (de serres, 1989c, 1990a). the array of genotypes in the classes and subclasses in the spectrum of ad-3 mutants induced by a mutagenic agent constitute its mutagenicity profile; for x-ra ...19902145510
cloning, sequence, and photoregulation of al-1, a carotenoid biosynthetic gene of neurospora crassa.carotenoid biosynthesis is regulated by blue light during growth of neurospora crassa mycelia. we have cloned the al-1 gene of n. crassa encoding the carotenoid-biosynthetic enzyme phytoene dehydrogenase and present an analysis of its structure and regulation. the gene encodes a 595-residue polypeptide that shows homology to two procaryotic carotenoid dehydrogenases. rna measurements showed that the level of al-1 mrna increased over 70-fold in photoinduced mycelia. transcription run-on studies i ...19902144609
molecular cloning and analysis of the scon-2 negative regulatory gene of neurospora crassa.the sulfur regulatory system of neurospora crassa is composed of a group of highly regulated structural genes (e.g., the gene encoding arylsulfatase) that are under coordinate control of scon+ (sulfur controller) negative and cys-3+ positive regulatory genes. in scon-1 (previously designated sconc) and scon-2 mutants, there is constitutive expression of sulfur structural genes regardless of the sulfur level available to the cells. the scon-2+ gene was cloned by sib selection screening of a cosmi ...19901975945
primary structure and expression of a nuclear-coded subunit of complex i homologous to proteins specified by the chloroplast genome.a 31-kda subunit of complex i from neurospora crassa, of nuclear origin, was cloned. the precursor polypeptide (33 kda) could be efficiently expressed in an in vitro system for transcription and translation. the processing of the precursor to the mature protein was also obtained in vitro. an open reading frame coding for a precursor protein of 283 amino acids (32247 da) was found by dna sequencing. the predicted primary structure shows significant homology with proteins made in chloroplast. this ...19902145832
the reaction of cn- with the binuclear copper site of neurospora tyrosinase: its relevance for a comparison between tyrosinase and hemocyanin active sites.the equilibrium and the kinetics of the reaction between neurospora crassa tyrosinase and cyanide have been studied. cyanide reacts with the binuclear copper active site of the protein competitively with respect to dioxygen and displaces the metal ions. this process occurs stepwise and involves transient intermediates containing mononuclear cu(i) sites. the reaction mechanism proved to be the same as described earlier for molluscan and arthropodan hemocyanins, which share with tyrosinase the sam ...19902145978
identification of the major cytoplasmic regions of the neurospora crassa plasma membrane h(+)-atpase using protein chemical techniques.the transmembrane topography of the neurospora crassa plasma membrane h(+)-atpase has been investigated using purified, reconstituted components and direct protein chemical techniques. reconstituted proteoliposomes containing h(+)-atpase molecules oriented predominantly with their cytoplasmic surface facing outward were treated with trypsin to liberate peptides present on the cytoplasmic surface of the h(+)-atpase as recently described (hennessey, j.p., jr., and scarborough, g. (1990) j. biol. c ...19902144525
nucleotide sequence and dna recognition elements of alc, the structural gene which encodes allantoicase, a purine catabolic enzyme of neurospora crassa.the nitrogen regulatory circuit of neurospora crassa contains structural genes that encode nitrogen catabolic enzymes which are subject to complex genetic and metabolic regulation. this set of genes is controlled by nitrogen limitation, by specific induction, and by the action of nit-2, a major positive-acting regulatory gene, and nmr, a negative-acting control gene. the complete nucleotide sequence of alc, the gene that encodes allantoicase, a purine catabolic enzyme, is presented. the alc gene ...19902148685
gene sequence and analysis of hsp30, a small heat shock protein of neurospora crassa which associates with mitochondria.hsp30 is a small heat shock protein of neurospora crassa which earlier studies suggested may associate with mitochondria during cellular heat shock. we show here that the association of hsp30 with mitochondria is reversible and that hsp30 dissociates after cells are returned to normal temperature. we sequenced the gene for hsp30 and defined its transcript by s1 nuclease analysis and cdna sequencing. the gene apparently is present in the genome as a single copy, and it contains no introns. the en ...19902144284
sorting pathways of mitochondrial inner membrane proteins.two distinct pathways of sorting and assembly of nuclear-encoded mitochondrial inner membrane proteins are described. in the first pathway, precursor proteins that carry amino-terminal targeting signals are initially translocated via contact sites between both mitochondrial membranes into the mitochondrial matrix. they become proteolytically processed, interact with the 60-kda heat-shock protein hsp60 in the matrix and are retranslocated to the inner membrane. the sorting of subunit 9 of neurosp ...19902145157
restricted activation of general amino acid control under conditions of glutamine limitation in neurospora crassa.in neurospora crassa limitation for single amino acids normally results in increased formation of enzymes required for amino acid synthesis via 'general amino acid control'. glutamine limitation, however, led to comparatively low and delayed derepression of enzyme synthesis. nitrate reductase activity increased steeply under these conditions confirming that de novo protein synthesis could occur. derepression levels were unaffected by addition of glutamine-derived metabolites. only small and dela ...19902148607
glutamine metabolism and cycling in neurospora crassa.evidence for the existence of a glutamine cycle in neurospora crassa is reviewed. through this cycle glutamine is converted into glutamate by glutamate synthase and catabolized by the glutamine transaminase-omega-amidase pathway, the products of which (2-oxoglutarate and ammonium) are the substrates for glutamate dehydrogenase-nadph, which synthesizes glutamate. in the final step ammonium is assimilated into glutamine by the action of a glutamine synthetase (gs), which is formed by two distinct ...19902145504
behavior of the [mi-3] mutation and conversion of polymorphic mtdna markers in heterokaryons of neurospora crassa.we have examined the behavior of the [mi-3] mitochondrial mutation and two physical mtdna markers in heterokaryotic cultures of neurospora crassa. previous workers showed that a 1.2-kilobase insertion in the larger polymorphic form of ecori-5 restriction fragment is a site of high frequency and rapid unidirectional gene conversion. we have confirmed this observation and determined by dna sequence analysis that the insertion in the ecori-5 fragment corresponds precisely to an optional intron that ...19901977658
isolation and characterization of a neurospora crassa mutant altered in the alpha polypeptide of glutamine synthetase.we report the isolation and characterization of a neurospora crassa glutamine synthetase (gs) mutant altered in one of the two polypeptides (gs alpha) of this enzyme. we used the gln-1br8 mutant strain that synthesizes only the gs alpha monomer and lacks the gs beta monomer and selected for growth in minimal medium in the presence of alpha-methyl-dl-methionine-sr-sulfoximine (alpha-me-mso), an inhibitor of gs activity. the gs activity of the gln-1br8;alpha-me-msor strain drastically reduced its ...19901975579
nucleotide sequence of a full-length cdna coding for the mitochondrial precursor protein of the beta-subunit of f1-atpase from neurospora crassa. 19902144339
effects of cell wall deficiency on the synthesis of polysaccharide-degrading exoenzymes: a study on mycelial and wall-less phenotypes of the fz; sg; os-1 ('slime') triple mutant of neurospora crassa.the production of exoenzymes which degrade cellulose, polygalacturonic acid and xylan was studied in mycelial and wall-less phenotypic derivatives of neurospora crassa obtained by vegetative selection applied to a single fz;sg;os-1 ('slime'-like) segregant (strain rcp-3) of a cross 'slime' x wild type. the unrelated stable 'slime' strain fgsc 1118 was also studied. the synthesis of polysaccharide-degrading enzymes was normally induced by polysaccharidic substrates and was sensitive to carbon-cat ...19902262786
x-ray-induced specific-locus mutations in the ad-3 region of two-component heterokaryons of neurospora crassa. vi. induction kinetics of gene/point mutations, multilocus deletions and multiple-locus mutations.genetic fine-structure analysis of x-ray-induced specific-locus mutants in the ad-3 region of two-component heterokaryons of neurospora crassa has shown that gene/point mutations, multilocus deletions and multiple-locus mutations are induced. when the dose-response curves for these classes of ad-3 mutants were plotted, it was demonstrated that x-ray-induced gene/point mutations (ad-3r) increased linearly with x-ray dose and x-ray-induced multilocus deletions increased as the square of the x-ray ...19902143556
biochemical, genetic and ultrastructural defects in a mitochondrial mutant (er-3) of neurospora crassa with senescence phenotype.the structural and functional abnormalities in a new respiratory deficient, mitochondrial senescence mutant er-3 of neurospora crassa are described. the mitochondrial mutant, which grows at a rate of only 10% of that of the wild type, was found deficient in all three cytochromes, and completely lacking in cytochromes aa3. cytochrome oxidase activity in the mutant mitochondria was only about 5% of the wild type mitochondria. however, the total whole cell respiration rate of the mutant was 33% gre ...19902169558
analysis of conventional and in vitro generated mutants of nmr, the negatively acting nitrogen regulatory gene of neurospora crassa.the nmr gene is the major negative regulatory gene in the nitrogen control circuit of neurospora crassa, which, together with positive regulatory genes, governs the expression of multiple unlinked structural genes of the circuit. possible functional domains of the nmr protein were investigated by mutational analyses using three different approaches. first, the polymerase chain reaction was used to clone the nmr locus from two conventional mutants, v2m304 and ms5, and the mutant amino acid codons ...19902148799
distant upstream regulatory sequences control the level of expression of the am (gdh) locus of neurospora crassa.we have constructed deletions in the 5' noncoding sequences of the cloned neurospora crassa am gene. vectors with a truncated fragment of the am gene were used in transformation experiments to introduce the deletions into the chromosome by homologous recombination. analysis of glutamate dehydrogenase (gdh) expression by enzyme assay and immunoblots, as well as northern and dot blots of poly (a)+ rna, in the deletion strains indicates that there are two upstream regulatory sequences that control ...19902147126
the cellulase complex of neurospora crassa: activity, stability and release.the temperature and ph optima, and the temperature and ph stability, of crude and purified enzymes of the cellulase complex of the cellulolytic ascomycete fungus neurospora crassa were investigated. the effects of some non-ionic surfactants and fatty acids on the production/release of enzymes of cellulase complex were also examined. for the different enzymes of the complex, activity maxima occurred between ph 4.0 and 7.0, with ph 5.0 being close to optimal for stability of all. temperature optim ...19902146364
assembly of nadh: ubiquinone reductase (complex i) in neurospora mitochondria. independent pathways of nuclear-encoded and mitochondrially encoded subunits.nadh:ubiquinone reductase, the respiratory chain complex i of mitochondria, consists of some 25 nuclear-encoded and seven mitochondrially encoded subunits, and contains as redox groups one fmn, probably one internal ubiquinone and at least four iron-sulphur clusters. we are studying the assembly of the enzyme in neurospora crassa. the flux of radioactivity in cells that were pulse-labelled with [35s]methionine was followed through immunoprecipitable assembly intermediates into the holoenzyme. la ...19902141652
a polypeptide of 59 kda is associated with bundles of cytoplasmic filaments in neurospora crassa.complex arrangements of filamentous structures have been isolated from vegetative cells of the fungus neurospora crassa. they were enriched by differential centrifugation and purified by permeation chromatography. the filamentous structures are made up of units of 8-10 nm diameter and were isolated in bundles of up to six to nine units. the main constituent of these structures is a polypeptide with an apparent molecular mass of 59 kda (p59nc), which represents 4-5% of the total n. crassa protein ...19902141976
intracellular sodium content of a wall-less strain of neurospora crassa and effects of insulin: a 23na-nmr study.23na-nmr has been used to investigate some factors influencing the sodium content of a wall-less strains of neurospora crassa. the shift reagent tm(dotp)h2(nh4)3 proved useful for this purpose, while several other reagents, previously used by others, were found to be unsuitable for use with these cells. when the cells were grown, washed and resuspended in medium containing sodium (25.3 mm), the intracellular sodium concentration was calculated to be 11.9 +/- 1.4 mm. this value rose within two mi ...19902142438
processing of precursor proteins by plant mitochondria.precursor proteins from neurospora crassa were correctly processed by a matrix extract from vicia faba and cauliflower mitochondria. processing yielded mature protein of the same molecular mass as mature neurospora protein. the processing activity has two components. one is antigenically related to and of the same molecular mass as the processing enhancing protein of neurospora. the second component was not recognized by antibody to the matrix processing protease from neurospora mitochondria. th ...19902140932
duplication-induced mutation of a new neurospora gene required for acetate utilization: properties of the mutant and predicted amino acid sequence of the protein product.a cloned neurospora crassa genomic sequence, selected as preferentially transcribed when acetate was the sole carbon source, was introduced in extra copies at ectopic loci by transformation. sexual crossing of transformants yielded acetate nonutilizing mutants with methylation and restriction site changes within both the ectopic dna and the normally located gene. such changes are typical of the duplication-induced premeiotic disruption (the rip effect) first described by selker et al. (e. u. sel ...19902140429
molecular cloning and characterization of alc the gene encoding allantoicase of neurospora crassa.purines can be utilized as a secondary nitrogen source by neurospora crassa during conditions of nitrogen limitation. the expression of purine catabolic enzymes is governed by the nitrogen regulatory circuit and requires induction by uric acid. the major positive-acting nitrogen regulatory gene, nit-2, turns on the expression of the purine catabolic enzymes, which may also be subject to negative regulation by a second control gene, nmr. we have cloned alc, the structural gene which encodes allan ...19901978237
chemical state of the cysteine residues in the neurospora crassa plasma membrane h(+)-atpase.the plasma membrane h(+)-atpase of neurospora crassa was treated with 5,5'-dithiobis(2-nitrobenzoate) to determine its cysteine content and with 2-nitro-5-thiosulfobenzoate to determine its cystine content. six and seven mol of thiols/mol of h(+)-atpase were detected in the 5,5'-dithiobis(2-nitrobenzoate) and 2-nitro-5-thiosulfobenzoate reactions, respectively, indicating that 6 of the 8 cysteine residues in the molecule are present as free cysteines and that 2 are present in disulfide linkage. ...19902139659
blue light photoreception in neurospora circadian rhythm: evidence for involvement of the flavin triplet state.the mechanism of the photoreceptor acting on the circadian conidiation rhythm of neurospora crassa was studied, with the following results: (1) the efficiency of 8-haloflavins as sensitizers increased with their triplet yields. (2) phase shifts were not abolished by removal of oxygen prior to illumination. (3) oxygen inhibited phase shifts when introduced into the cultures after light treatment. it is proposed that the blue light photoreceptor for the circadian clock of neurospora crassa acts (1 ...19902367558
development of thermotolerance in neurospora crassa by heat shock and other stresses eliciting peroxidase induction.hyperthermia, cdcl2, sodium arsenite, and h2o2 led to the rapid appearance of high levels of peroxidase in neurospora crassa cultures and induced tolerance toward normally lethal temperatures in 60-h-old colonies. intracellular superoxide dismutase levels did not correlate with the development of thermotolerance.19902139653
the genetics of polyamine synthesis in neurospora crassa.new mutations of the polyamine pathway of neurospora crassa fell into three categories. the majority affected ornithine decarboxylase and lay at the previously defined spe-1 locus. one mutation, jp100, defining the new spe-2 locus, eliminated s-adenosyl-methionine decarboxylase and led to putrescine accumulation. revertants of this mutation suggested that the locus encodes the enzyme. two other mutations, lv105 and jp120, defined a third locus, spe-3. strains with these mutations also accumulate ...19902139316
characterization of telomere dna from neurospora crassa.the nucleotide sequence of the telomere at the right end of linkage group v (vr) in the standard or23-iv-a strain of the filamentous fungus, neurospora crassa, reveals the following features. at the chromosome terminus, tandem repeats of the hexanucleotide ttaggg are present. immediately centromere-proximal to the simple sequence repeat is a more complex element called pogo that is reiterated 5-10 times in the genomes of various neurospora strains. the element possesses several features characte ...19901971801
light-induced dephosphorylation of a 33 kda protein in the wild-type strain of neurospora crassa: the regulatory mutants wc-1 and wc-2 are abnormal.light induces the dephosphorylation of a 33 kdalton protein within 8 min in the wild-type strain of neurospora crassa. the regulatory mutants, wc-1 and wc-2, have an altered pattern of phosphoproteins in darkness and also after irradiation. because the wc genes have previously been implicated in photodifferentiation (f. degli innocenti and v. e. a. russo, genetic analysis of blue light-induced responses in neurospora crassa, in h. senger (ed.), blue light effects in biological systems, springer- ...19902140412
very low atp/adp ratios with aging of the natural death senescence mutant of neurospora crassa.the natural death (nd) mutant of the fungus neurospora crassa, unlike the wild-type, undergoes an aging process, which leads to the cessation of growth. it is shown here that the atp/adp ratio of the mutant declines with age to about 3:1 whereas other strains of neurospora in the same growth medium maintain ratios of about 8 to 9:1. the decline in atp/adp ratio is not caused by the cessation of growth of the mutant. the results suggest, rather, that the cessation of growth may be caused, in part ...19902139154
relationship of vector insert size to homologous integration during transformation of neurospora crassa with the cloned am (gdh) gene.we used lambda and plasmid vectors containing the am+ gene in an insert of from 2.7 to 9.1 kb, to transform am point mutant and deletion strains. a total of 199 transformants were examined with the potential to yield am+ transformants by homologous recombination. when we used vectors that had 9.1 kb of homology with the chromosomal dna, 30% of the transformants obtained were the result of homologous recombination regardless of whether the vector was a lambda molecule, a circular plasmid, or a pl ...19902157957
dna methylation and chromatin structure: a view from below.an understanding of the function and control of dna methylation in eukaryotes has been elusive. studies of neurospora crassa have led to a model that accounts for the chromosomal distribution of methylation and suggests a basic function for dna methylation in eukaryotes.19902139257
determination of the inactivating alterations in two mutant alleles of the neurospora crassa cross-pathway control gene cpc-1.cpc-1 is the locus specifying what is believed to be the major trans-activating transcription factor that regulates expression of amino acid biosynthetic genes subject to cross-pathway control in neurospora crassa. mutants altered at this locus are incapable of the global increase in gene expression normally seen in response to amino acid starvation. using polymerase chain reaction methodology we have cloned and sequenced the inactive mutant allele, cpc-1 (cd15). the cpc-1 (cd15) mutation was fo ...19902138111
synaptic adjustment of inversion loops in neurospora crassa.heterozygotes for three long inversions on chromosome 1 were analyzed by serial reconstruction from electron micrographs. measurements of loop lengths at different meiotic prophase substages revealed that the homologous synapsis of the inverted region was gradually replaced by nonhomologous synapsis as loops were eliminated during pachytene. this synaptic adjustment was apparently not affected by crossovers which occurred within the 150- and 160-cm long loops.19902138110
nit-2, the major nitrogen regulatory gene of neurospora crassa, encodes a protein with a putative zinc finger dna-binding domain.the nitrogen regulatory circuit of neurospora crassa consists of a set of unlinked structural genes which specify various nitrogen catabolic enzymes plus control genes and metabolic effectors which regulate their expression. the positive-acting nit-2 regulatory gene is required to turn on the expression of the nitrogen catabolic enzymes during conditions of nitrogen limitation. the complete nucleotide sequence of the nit-2 gene was determined. the nit-2 mrna is 4.3 kilobases long and has a long ...19902137552
cis,cis-cyclohexane 1,3,5-triol polyphosphates release calcium from neurospora crassa via an unspecific ins 1,4,5-p3 receptor.we investigated the effects of new inositol 1,4,5-trisphosphate analogues on the release of ca2+ from isolated vacuoles of neurospora crassa. tri-o-butyryl-inositol 1,4,5-trisphosphate and a set of cis,cis-cyclohexane 1,3,5-triol bis-(cht-p2) and trisphosphates (cht-p3) gave an increase in free ca2+ as measured directly with fura-2, a ca2(+)-chelator. however, inositol 1,4-bisphosphate, 6-o-palmitoyl-inositol 4,5-bisphosphate and trans-cyclohexane 1,2-diol bisphosphate (trans chd-p2) did not ind ...19902154977
the range of amino acids whose limitation activates general amino-acid control in neurospora crassa.several amino-acid synthetic enzymes, belonging to arginine, glutamine, leucine, lysine and phenylalanine biosynthesis, respectively, were investigated under conditions of reduced availability of any one of 16 out of the 20 amino acids represented in proteins. the enzymes showed simultaneous derepression under each condition, albeit to different degrees. derepression was abolished and the remaining basal enzyme levels reduced by mutations at the cpc-1 locus which governs general amino-acid contr ...19902138581
cellulase production by neurospora crassa: purification and characterization of cellulolytic enzymes.in studies on cellulase production by the cell-1 mutant of neurospora crassa, eight enzymes (three exoglucanases, four endoglucanases, and one beta-glucosidase) were identified and characterized by gel filtration, ion exchange chromatography, and chromatofocusing. after purification, each of the proteins ran as a single band in polyacrylamide gel electrophoresis, using both native and denaturing gels. the molecular weights of the proteins were found to be between 70,000 and 22,000 daltons, and a ...19901368541
de novo fatty acid synthesis mediated by acyl-carrier protein in neurospora crassa mitochondria.the acyl-carrier protein (acp) in neurospora crassa mitochondria [brody, s. & mikolajczyk, s. (1988) eur. j. biochem. 173, 353-359] mediated a cerulenin-sensitive, de novo fatty acid synthesis independent of the fatty acid synthetase complex present in the cytoplasm. incubation of mitochondria with [2-14c]malonate labeled only the acp as indicated by autoradiography after sds/page. under these in vitro conditions atp was required for the initial acyl-acp formation, but further elongation require ...19902137086
nucleotide sequence of the gene coding for cyclophilin/peptidyl-prolyl cis-trans isomerase of neurospora crassa. 19902137907
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