PMID(sorted descending)
characterization of the ugata gene of ustilago maydis, isolated by homology to the gata gene of aspergillus nidulans.a gene encoding a putative gaba aminotransferase (ugata) was isolated from the basidiomycete ustilago maydis via heterologous hybridization to the gaba aminotransferase gene (gata) of aspergillus nidulans . the derived amino-acid sequence of ugata shows strong identity throughout the protein to the gaba aminotransferase enzymes from a. nidulans and saccharomyces cerevisiae. northern analysis in u. maydis indicated that the ugata transcript is inducible by the omega-amino acids gaba and beta-alan ...19968598057
cata, a new aspergillus nidulans gene encoding a developmentally regulated catalase.aspergillus nidulans asexual sporulation (conidiation) is a model system for studying gene regulation and development. the can5 cdna is one of several clones isolated based on transcript induction during conidiation. here we present the molecular characterization of its corresponding gene, demonstrating that it encodes a developmentally regulated catalase, designated cata. the cata 744-amino-acid-residue polypeptide shows significant identity to other catalases. its similarity to prokaryotic cat ...19968598056
mutational analysis reveals dispensability of the n-terminal region of the aspergillus transcription factor mediating nitrogen metabolite repression.mutational analysis has enabled identification and localization of an upstream exon of the area gene of aspergillus nidulans mediating nitrogen metabolite repression. a mutation in the initiation codon and frameshift mutations, which revert by restoration of the reading frame, established the coding role of the exon and mutations affecting intron splicing in conjunction with dna sequencing of reverse transcriptase polymerase chain reaction (rt-pcr) products localized the coding region intron. th ...19958596437
wild chromosomal variants in aspergillus nidulans.pulsed-field gel electrophoresis and a chromosome-specific cosmid dna library were used to determine the karyotypes of wild-type aspergillus nidulans isolates from around the world. overall, little structural variation was found, with a few major exceptions. one isolate possessed a non-essential b-chromosome of about 1.0 million base pairs (mb). another isolate had undergone a non-reciprocal translocation of about 1.6 mb of chromosome vi onto chromosome viii. other than these chromosomal differe ...19968595677
two family g xylanase genes from chaetomium gracile and their expression in aspergillus nidulans.with oligonucleotides based on the amino-terminal and internal amino-acid sequences of a xylanase, two xylanase genes, cgxa and cgxb, were isolated and sequenced from chaetomium gracile wild and mutant strains. each gene isolated from both strains was essentially the same as far as nucleotide sequences were compared. the mature cgxa and cgxb xylanases comprise 189 and 211 amino acids, respectively, and share 68.5% homology. the cgxa was found to be the major enzyme in the mutant strain. comparis ...19958595661
biolistic transformation of the obligate plant pathogenic fungus, erysiphe graminis f.sp. hordei.particle gun acceleration appears to be a possible way to transform mycelium cells of obligate plant parasites growing on host surfaces. gus expression was obtained in e. graminis f.sp. hordei cells after bombardment with the gus gene under the control of the e. graminis f.sp. hordei β-tubulin promoter. three heterologous promoters, onefrom aspergillus nidulans and two from cochliobolus heterostrophus, gave very low or no expression of gus.19958595653
transformations of penicillium islandicum and penicillium frequentans that produce anthraquinone-related compounds.wild-type strains of penicillium islandicum and penicillium frequentans, which produce anthraquinone and related compounds, were transformed to benomyl and hygromycin b resistance. plasmids psv50 and pbt6, with benomyl-resistant beta-tublin genes, and plasmids pan7-1 and pdh25, with a bacterial hygromycin phosphotransferase gene under the control of aspergillus nidulans sequences, were used respectively. transformation frequencies with these plasmids were 10-20 transformants per micrograms of dn ...19958593690
organisation of the mitochondrial genome of trichophyton rubrum. dna sequence analysis of the nd4 gene, the atpase subunit-6 gene, the ribosomal rna small-subunit gene, the nd6 gene, the coxiii gene, the atpase subunit-8 gene and six trna genes that correspond respectively to the tyrosine, lysine, glutamine, asparagine, isoleucine and tryptophan isoacceptors.we present the nucleotide sequence of a 5207-bp-long region of the mitochondrial genome of the dermatophyte trichophyton rubrum. this represents about 1/5th of the total genome and extends a previous study. from the 5' end of the present sequence, the order of genes is as follows: the end of the nd4 gene, the gene coding for subunit 6 of atpase, the gene coding for the small ribosomal rna (ssu rrna), the tyrosyl trna gene, the nd6 gene, the coxiii gene, the atpase 8 subunit gene and a cluster of ...19958593686
generation and characterization of nadh: ubiquinone oxidoreductase mutants in neurospora crassa. 19958592454
optimization of glucose oxidase production by aspergillus niger using genetic- and process-engineering techniques.wild-type aspergillus niger nrrl-3 was transformed with multiple copies of the glucose oxidase structural gene (god). the gene was placed under the control of the gpda promoter of a. nidulans. for more efficient secretion the alpha-amylase signal peptide from a. oryzae was inserted in front of god. compared to the wild type, the recombinant strain nrrl-3 (god3-18) produced up to four times more extracellular glucose oxidase under identical culture conditions. addition of yeast extract (2 gl-1) t ...19958590664
pneumonia due to fonsecaea pedrosoi and cerebral abscesses due to emericella nidulans in a bone marrow transplant recipient. 19958589181
the aspergillus nidulans bime (blocked-in-mitosis) gene encodes multiple cell cycle functions involved in mitotic checkpoint control and mitosis.the bime (blocked-in-mitosis) gene appears to function as a negative mitotic regulator because the recessive bime7 mutation can override certain interphase-arresting treatments and mutations, causing abnormal induction of mitosis. we have further investigated the role of bime in cell cycle checkpoint control by: (1) coordinately measuring mitotic induction and dna content of bime7 mutant cells; and (2) analyzing epistasis relationships between bime7 and 16 different nim mutations. a combination ...19958586660
domain structure and function within the quta protein of aspergillus nidulans: implications for the control of transcription.quta is a positively acting regulatory protein that regulates the expression of the eight genes comprising the quinic acid utilization gene (qut) gene cluster in aspergillus nidulans. it has been proposed that the quta protein is composed of two domains that are related to the n-terminal two domains-dehydroquinate (dhq) synthase and 5-enolpyruvyl shikimate-3-phosphate (epsp) synthase-of the pentadomain arom protein. the arom protein is an enzyme catalysing five consecutive steps in the shikimate ...19968581174
ethanol utilization regulatory protein: profile alignments give no evidence of origin through aldehyde and alcohol dehydrogenase gene fusion.the suggestion that the ethanol regulatory protein from aspergillus has its evolutionary origin in a gene fusion between aldehyde and alcohol dehydrogenase genes (hawkins ar, lamb hk, radford a, moore jd, 1994, gene 146:145-158) has been tested by profile analysis with aldehyde and alcohol dehydrogenase family profiles. we show that the degree and kind of similarity observed between these profiles and the ethanol regulatory protein sequence is that expected from random sequences of the same comp ...19958580855
integrative and replicative transformation of penicillium canescens with a heterologous nitrate-reductase gene.a wild isolate of penicillium canescens was subjected to mutagenesis, and 150 chlorate-resistant mutants were isolated and classified in respect of their ability to utilize various nitrogen sources. strains supposedly deficient in nitrate reductase have been transformed with the nitrate-reductase gene from aspergillus niger. transformation probably occurred by non-homologous integration of the transforming vector into the chromosome. co-transformation with the ama 1 replicating element from a. n ...19958575022
biosynthetic pathways of glycerol accumulation under salt stress in aspergillus nidulans.a culture of aspergillus nidulans (fgsc 359) was gradually adapted for growth in media containing up to 2 m nacl or was exposed to a salt shock with 2 m nacl. the intracellular glycerol level increased by about 7.9-fold in salt-adapted and 2.4-fold in salt-shocked cultures when compared to the unadapted culture. the biosynthetic pathway involved in the accumulation of glycerol was investigated under long-term salt adaptation and short-term salt shock. glycerol-3-phosphate dehydrogenase (ec 1.1.1 ...19958574901
allocation of random amplified polymorphic dna markers and enzyme activities to aspergillus nidulans and aspergillus tetrazonus chromosomes.chromosome-substituted haploid segregants of an a. nidulans x a. tetrazonus somatic hybrid were used to allocate several random amplified polymorphic dna and isoenzyme markers to parental chromosomes. twenty-six amplified dna fragments, and nine isoenzyme activities, including lactate dehydrogenase, superoxide dismutase, and arylesterase isoenzymes were assigned to chromosomes. chromosomes-specific markers were found for each a. nidulans and a. tetrazonus chromosome. these markers could be used ...19958572683
mutational analysis of the c-terminal region of area, the transcription factor mediating nitrogen metabolite repression in aspergillus aspergillus nidulans the positive-acting, wide domain regulatory gene area mediates nitrogen metabolite repression. previous analysis demonstrated that the c-terminal 153 residues of the area product (area) are inessential for at least partial expression of most genes subject to regulation by area. paradoxically, arear2, a -1 frameshift replacing the wild-type 122 c-terminal residues with a mutant peptide of 117 amino acids, leads to general loss of function. to determine the basis for the ar ...19968569680
enzymatic synthesis of hydrophobic penicillins.our knowledge of the enzymes and genes involved in the biosynthesis of beta-lactam antibiotics has increased notably in the last decade. the purification to homogeneity of some of these proteins as well as their biochemical characterization has allowed some of them to be used for synthesizing many different penicillins and cephalosporin-like products in vitro. in this report we describe the most important advances in this field, placing special emphasis on the enzymatic synthesis of hydrophobic ...19958557558
structure-function analysis of the proline permease (prnb) of the filamentous fungus aspergillus nidulans. 19948550020
genetic and molecular characterisation of purine permease genes of aspergillus nidulans reveals a novel family of transporters conserved in prokaryotes and eukaryotes.the ascomycete fungus aspergillus nidulans can utilize purines (adenine, guanine, hypoxanthine, xanthine, and uric acid) as sole nitrogen sources [1]. the expression of most structural genes involved in the pathway of purine uptake and catabolism is subject to uric acid induction, mediated by the product of the positive regulatory gene uay, and to nitrogen metabolite repression, mediated by the product of the general, positive-acting, gata-like transcription factor, encoded by the area gene [1].19948550005
transport of lactate and its regulation in saccharomyces cerevisiae mutants deficient in specific metabolic steps. 19948550003
toxicology of halogenated aliphatic hydrocarbons: structural and molecular determinants for the disturbance of chromosome segregation and the induction of lipid peroxidation.the induction of mitotic chromosome malsegregation, mitotic arrest and lethality by a set of 55 halogenated hydrocarbons was investigated. to this aim, genetic assays in the mould aspergillus nidulans, able to provide precise quantitative information on the end-points studied, were used throughout the work. the experimental data obtained were used to develop qsar models for the induction of aneuploidy, which pointed to a major role of electrophilicity as molecular determinant for the aneugenic p ...19958548852
cell cycle. the nima kinase joins forces with cdc2.the nima and cdc2 protein kinases cooperate to regulate mitosis in aspergillus nidulans. nima-related pathways have now begun to emerge in higher eukaryotes.19958548283
analysis of the regulation of the aspergillus nidulans penicillin biosynthesis gene aat (pende), which encodes acyl coenzyme a:6-aminopenicillanic acid acyltransferase.the regulation of the aspergillus nidulans penicillin biosynthesis gene aat (pende), which encodes acyl coenzyme a:6-aminopenicillanic acid acyltransferase (aat), was analysed. major transcriptional start sites map within 100 nucleotides upstream from the aat initiation codon. to study the regulation of aat expression, various aat-lacz gene fusions were constructed, in which the aat promoter region was fused in frame with the escherichia coli lacz reporter gene. a. nidulans strains carrying reco ...19958544821
quantification of dna damage and repair in amino acid auxotrophs and uv-sensitive mutants of aspergillus nidulans using an elisa used to investigate dna repair in mammalian cells has been adapted to investigate mutagen-induced dna damage and repair in protoplasts of aspergillus nidulans. the assay shows a reduced rate of repair of dna damage in methionine and arginine auxotrophs (methg and argb), which were shown previously to be hypersensitive to uv radiation and chemical mutagens. the assay also showed a considerably reduced ability to repair mutagen-induced damage in the uv-sensitive mutants uvsb and uvsh. the ...19958543032
recombinational stability of replicating plasmids in aspergillus nidulans during transformation, vegetative growth and sexual reproduction.plasmids containing the ama1 replicon are capable of autonomous maintenance in aspergillus nidulans. it has been reported previously that these plasmids can form concatenates by recombination in a transformed mycelium, and up to 10% of molecules are involved in such events. the present study demonstrates that plasmid recombination, although frequent during transformation, rarely occurs during vegetative growth. as a result, the structure and phenotypic stability of ama1 plasmids generally remain ...19958536318
characterization of the "promoter region" of the enolase-encoding gene enol from the anaerobic fungus neocallimastix frontalis: sequence and promoter analysis.the sequence of the neocallimastix frontalis enolase gene promoter was determined up to 1800 nucleotides 5' to the major transcriptional start point. the base composition of the enolase upstream sequence revealed a very a + t-rich profile (13.5% g + c) leading to many putative hairpin structures. the functional organization of the n. frontalis enolase promoter was investigated by heterologous transient-expression assays. dna fragments obtained by the sequential removal of sequences upstream of t ...19958536317
gaip, a protein that specifically interacts with the trimeric g protein g alpha i3, is a member of a protein family with a highly conserved core domain.using the yeast two-hybrid system we have identified a human protein, gaip (g alpha interacting protein), that specifically interacts with the heterotrimeric gtp-binding protein g alpha i3. interaction was verified by specific binding of in vitro-translated g alpha i3 with a gaip-glutathione s-transferase fusion protein. gaip is a small protein (217 amino acids, 24 kda) that contains two potential phosphorylation sites for protein kinase c and seven for casein kinase 2. gaip shows high homology ...19958524874
sok2 may regulate cyclic amp-dependent protein kinase-stimulated growth and pseudohyphal development by repressing transcription.yeast cyclic amp (camp)-dependent protein kinase (pka) activity is essential for growth and cell cycle progression. dependence on pka function can be partially relieved by overexpression of a gene, sok2, whose product has significant homology with several fungal transcription factors (stua from aspergillus nidulans and phd1 from saccharomyces cerevisiae) that are associated with cellular differentiation and development. deletion of sok2 is not lethal but exacerbates the growth defect of strains ...19958524252
role of tryptophans in substrate binding and catalysis by dna photolyase. 19958524158
dictyostelium myosin i double mutants exhibit conditional defects in pinocytosis.the functional relationship between three dictyostelium myosin is, myoa, myob, and myoc, has been examined through the creation of double mutants. two double mutants, myoa-/b- and myob-/c-, exhibit similar conditional defects in fluid-phase pinocytosis. double mutants grown in suspension culture are significantly impaired in their ability to take in nutrients from the medium, whereas they are almost indistinguishable from wild-type and single mutant strains when grown on a surface. the double mu ...19958522584
cre1, the carbon catabolite repressor protein from trichoderma order to investigate the mechanism of carbon catabolite repression in the industrially important fungus trichoderma reesei, degenerated pcr-primers were designed to amplify a 0.7-bp fragment of the cre1 gene, which was used to clone the entire gene. it encodes a 402-amino acid protein with a calculated m(r) of 43.6 kda. its aa-sequence shows 55.6% and 54.7% overall similarity to the corresponding genes of aspergillus nidulans and a. niger, respectively. similarity was restricted to the aa-reg ...19958521952
the nitrate reductase gene from a shoyu koji mold, aspergillus oryzae kbn616.a niad gene encoding nitrate reductase was isolated from aspergillus oryzae kbn616 and sequenced. the structural gene comprises 2973 bp and 868 amino acids, which showed a high degree of similarity to nitrate reductases from other filamentous fungi. the coding sequence is interrupted by six introns varying in size from 48 to 98 bp. the intron positions are all conserved among the niad genes from a. oryzae, aspergillus nidulans, and aspergillus niger. a homologous transformation system was develo ...19958520125
estimation of mitotic stability in conidial fungi: a theoretical framework.mitotic stability refers to the probability that genetic elements are transmitted to both daughters during mitosis. this is of practical importance in molecular genetics because autonomous cloning vectors should be transmitted at high frequency during mitosis. in filamentous coencytic fungi it is difficult to quantify mitotic stability because a fluctuation test is not feasible. we show how to get around this problem by formulating a general model of the transmission of nuclear genetic elements ...19938514143
translational repression of brla expression prevents premature development in aspergillus.the aspergillus nidulans brla developmental regulatory locus consists of two overlapping transcription units, brla alpha and brla beta, which encode functionally related polypeptides. we used translational fusions between each of the predicted brla reading frames and the escherichia coli lacz gene to test the hypothesis that developmental regulation of brla alpha and brla beta expression occurs through different mechanisms. brla alpha is transcriptionally controlled and a large portion of brla a ...19938508770
the aspergillus nidulans brla regulatory locus consists of overlapping transcription units that are individually required for conidiophore development.the aspergillus nidulans brla locus controls conidiophore development in conjunction with the products of several other regulatory loci. in this paper, we show that the brla locus consists of overlapping transcription units, designated alpha and beta, with alpha transcription initiating within beta intronic sequences. the predicted brla polypeptides differ by 23 amino acid residues at their n-termini. targeted mutations specifically eliminating either the alpha or beta transcript led to developm ...19938508769
the aspergillus nidulans ya gene is regulated by abaa.the developmentally regulated aspergillus nidulans ya gene encodes a p-diphenol oxidase that is needed for synthesis of green conidial pigment. we subjected the ya 5' flanking region to mutational analysis in a. nidulans and saccharomyces cerevisiae to identify dna sequence elements involved in its transcriptional control, and identified two functionally distinct elements. element i contained potential brla binding sites and was required for full level ya transcription, but not for developmental ...19938491194
essential roles for calcium and calmodulin in g2/m progression in aspergillus nidulans.nimt encodes a protein in aspergillus nidulans that is required for tyrosine dephosphorylation of p34cdc2 and has a strong homology to cdc25-type proteins. conditional mutation of nimt (nimt23 mutation) arrests cells in g2 at the restrictive temperature. after release of the temperature-sensitive nimt23 block, p34cdc2 undergoes tyrosine dephosphorylation and we showed that as cells entered mitosis, a rapid increase in calmodulin was observed. the increase in calmodulin and progression into mitos ...19938486741
specific binding sites in the alcr and alca promoters of the ethanol regulon for the crea repressor mediating carbon catabolite repression in aspergillus nidulans.the crea repressor responsible for carbon catabolite repression in aspergillus nidulans represses the transcription of the ethanol regulon. the n-terminal part of the crea protein encompassing the two zinc fingers (c2h2 class family) and an alanine-rich region was expressed in escherichia coli as a fusion protein with glutathione-s-transferase. our results show that crea is a dna-binding protein able to bind to the promoters of both the specific trans-acting gene, alcr, and of the structural gen ...19938483416
at least four regulatory genes control sulphur metabolite repression in aspergillus nidulans.mutations in four genes: scona (formerly sua25meth, mapa25), sconb (formerly mapb1), sconc and scond, the last two identified in this work, relieve a group of sulphur amino acid biosynthetic enzymes from methionine-mediated sulphur metabolite repression. exogenous methionine has no effect on sulphate assimilation in the mutant strains, whereas in the wild type it causes almost complete elimination of sulphate incorporation. in both mutant and wild-type strains methionine is efficiently taken up ...19938479426
properties of genes involved in the control of isocitrate lyase production in aspergillus nidulans.the interaction between genes of aspergillus nidulans conferring constitutive synthesis of isocitrate lyase (iclc a and iclcb) and fluoroacetate resistance (facb) has been investigated. although facb mutants are unable to induce the glyoxylate cycle enzyme isocitrate lyase in response to acetate as sole carbon source, this phenotype was suppressed in recombinants of the type iclc;facb. the iclca and iclcb mutations do not alter significantly the activities of eight enzymes of intermediary metabo ...19938473860
properties and regulation of the cell cycle-specific nima protein kinase of aspergillus nidulans.nima is the protein product of the nima gene of the filamentous fungus aspergillus nidulans, required for progression of cells from g2 into mitosis. the protein kinase activity of nima, assayed by phosphorylation of beta-casein, varies during the nuclear division cycle, reaching a maximum in late g2 and m. to investigate the biochemical properties of this cell cycle-regulated protein kinase, we have expressed nima cdna that encodes full-length nima in escherichia coli as a fusion product with gl ...19938473320
carnitine acetyltransferase is absent from acuj mutants of aspergillus nidulans.the acuj mutant of aspergillus nidulans has been shown to lack carnitine acetyltransferase (cat) activity when grown under conditions where this activity is readily detectable in wild-type strains. revertants selected for growth on acetate recover cat activity and the ability to grow on long-chain fatty acids. when growing on carbon sources such as sucrose, cytosolic acetyl coenzyme a was generated by adenosine triphosphate (atp):citrate lyase.19938472927
evolutionary conservation of excision repair in schizosaccharomyces pombe: evidence for a family of sequences related to the saccharomyces cerevisiae rad2 gene.cells mutated at the rad13 locus in the fission yeast, schizosaccharomyces pombe are deficient in excision-repair of uv damage. we have cloned the s.pombe rad13 gene by its ability to complement the uv sensitivity of a rad13 mutant. the gene is not essential for cell proliferation. sequence analysis of the cloned gene revealed an open reading-frame of 1113 amino acids with structural homology to the rad2 gene of the distantly related saccharomyces cerevisiae. the sequence similarity is confined ...19938464724
aspergillus nidulans nuclear proteins bind to a ccaat element and the adjacent upstream sequence in the promoter region of the starch-inducible taka-amylase a gene.aspergillus nidulans was used as an intermediate host to investigate the regulation of the taka-amylase a (taa) gene from aspergillus oryzae. the induction of taa by starch was confirmed to be regulated at the transcriptional level by analyzing the transcripts specific for taa synthesized in vitro in nuclei from starch- and glucose-grown cells. a 55 bp dna fragment containing a consensus ccaat sequence from the promoter region of the taa gene was shown to confer starch inducibility on the gene. ...19938455560
cerato-ulmin, a toxin involved in dutch elm disease, is a fungal hydrophobin. 19938453298
the mitochondrial genome of the entomopathogenic fungus beauveria bassiana: analysis of the ribosomal rna region.the 28.5-kbp mitochondrial (mt) genome from the entomopathogenic fungus beauveria bassiana was studied using restriction enzyme analysis, gene probe hybridization, and dna sequence comparisons. a detailed restriction enzyme map allowed cloning of the entire genome into a number of segments. hybridization of heterologous gene probes to the mtdna resulted in the identification of the large ribosomal rna (lrrna) and small ribosomal rna (srrna) genes. gene probes derived from several yeasts and fung ...19938439871
molecular cloning and analysis of abundant and stage-specific mrnas from puccinia characterize highly expressed mrnas from germinated urediniospores of puccinia graminis f. sp. tritici, we isolated 68 cdna clones of abundant mrna species belonging to at least six homology groups. the two most abundant homology groups, hg1 and hg2, contained 54 of the 68 cdna clones and accounted for 2.4 and 0.6% of the poly(a)+ rna in germinated urediniospores, respectively. by sampling different developmental stages of the uredinial cycle, we showed that the uam transcript, corresponding ...19938439672
cloning of two isozymes of trichoderma koningii glyceraldehyde-3-phosphate dehydrogenase with different sensitivity to koningic acid.koningic acid inhibits glyceraldehyde-3-phosphate dehydrogenase (gapdh) by binding to the sh group in the active center. the fungus trichoderma koningii, the producer of koningic acid, contains two gapdh isozymes (gapdhs i and ii). gapdh i is inhibited 50% by 1.1.10(-3) m koningic acid, while gapdh ii is inhibited 50% at 6.8 x 10(-6) m. cdnas of the two isozymes were cloned from t. koningii and their nucleotide sequences were determined. the sequence of coding region and codon usage in both clon ...19938439569
operator derepressed mutations in the proline utilisation gene cluster of aspergillus nidulans.the proline utilisation gene cluster of aspergillus nidulans can be repressed efficiently only when both repressing nitrogen and repressing carbon sources are present. we show that two cis-acting mutations in this cluster permit the efficient transcription of the prnb gene under repressing conditions, resulting in direct or indirect derepression of two other transcripts of the pathway. these mutations are transitions that define a 5'gagacccc3' sequence. similar sequences are found upstream of ot ...19938437566
c-terminal truncation of the transcriptional activator encoded by area in aspergillus nidulans results in both loss-of-function and gain-of-function phenotypes.mutations truncating as many as 143 c-terminal residues from the transcriptional activator encoded by the area gene, mediating nitrogen metabolite repression in aspergillus nidulans, do not significantly reduce the ability of the area product to activate expression of most genes under area control. such mutations can even have a gain-of-function, derepressed phenotype, consistent with a critical role for this region in modulating the activity of the area protein. however, expression of a few gen ...19938437521
bima, a tpr-containing protein required for mitosis, localizes to the spindle pole body in aspergillus nidulans.the aspergillus nidulans bima gene is required for mitosis. loss of function mutations in bima cause cells to arrest growth with condensed chromatin and a short, metaphaselike mitotic spindle. bima is a member of a gene family defined by a repeated motif called the tetratrico peptide repeat (tpr), which is found in genes from bacteria, yeast and insects. several yeast tpr genes are also required for mitosis, including saccharomyces cerevisiae cdc27 and schizosaccharomyces pombe nuc2+, which appe ...19938432735
cloning and sequencing of the 3-phosphoglycerate kinase (pgk) gene from penicillium citrinum and its application to heterologous gene expression.the gene coding for 3-phosphoglycerate kinase (pgk) in ml-236b (compactin)-producing penicillium citrinum was isolated from the recombinant phage lambda library using the corresponding aspergillus nidulans pgk gene as a probe. the p. citrinum pgk gene has an open reading frame of 1,254 bp, encoding a protein of 417 amino acids with a predicted molecular weight of 44,079 daltons. the position of the two introns, 59 and 60 bp respectively, was deduced from an homology comparison with the sequence ...19938431954
disruption of the 3' phosphoglycerate kinase (pgk) gene in aspergillus nidulans.we report the isolation of a pgk- mutant strain of aspergillus nidulans by means of a gene disruption strategy, and demonstrate that the pgk gene is located on chromosome viii. the pgk- mutant conidiates poorly, will only grow on media supplemented with both a glycolytic and a gluconeogenic carbon source, and is inhibited by hexoses.19938431952
genetic maps of eight linkage groups of aspergillus niger based on mitotic mapping.this paper provides a genetic map of aspergillus niger. at present 84 markers have been assigned to eight linkage groups. the chromosomal location of 60 markers is presented in this paper. the allocation of markers is based on recombination due to mitotic crossing over. various methods for selection and analysis of homozygous recombinants were applied, using colour, auxotrophic and resistance markers. in addition, transformants carrying the heterologous aspergillus nidulans gene coding for aceta ...19938428383
identification of aspergillus brla response elements (bres) by genetic selection in yeast.the brla gene of aspergillus nidulans plays a central role in controlling conidiophore development. to test the hypothesis that brla encodes a transcriptional regulator and to identify sites of interaction for the brla polypeptide, we expressed brla in saccharomyces cerevisiae (yeast) strains containing aspergillus dna sequences inserted upstream of a minimal yeast promoter fused to the escherichia coli lacz gene. initially, a dna fragment from the promoter region of the developmentally regulate ...19938417986
suppression of the bimc4 mitotic spindle defect by deletion of klpa, a gene encoding a kar3-related kinesin-like protein in aspergillus investigate the relationship between structure and function of kinesin-like proteins, we have identified by polymerase chain reaction (pcr) a new kinesin-like protein in the filamentous fungus aspergillus nidulans, which we have designated klpa. dna sequence analysis showed that the predicted klpa protein contains a cooh terminal kinesin-like motor domain. despite the structural similarity of klpa to the kar3 and ncd kinesin-like proteins of saccharomyces cerevisiae and drosophila melanogaste ...19938416986
functional redundancy in mitotic force generation. 19938416980
biochemical characterization and molecular genetics of nine mutants of penicillium chrysogenum impaired in penicillin biosynthesis.nine mutants of penicillium chrysogenum (npe1 to npe8 and npe10) impaired in penicillin biosynthesis were screened after nitrosoguanidine mutation. mutants npe1, npe4, npe5, npe6, npe7, npe8, and npe10 failed to synthesize significant levels of penicillin, whereas strains npe2 and npe3 synthesized about 20% of the penicillin level produced by the parental strain. mutants npe5 and npe10 did not show alpha-aminoadipylcysteinyl-valine (acv) synthetase activity in vitro and did not form acv in vivo. ...19938416976
purification and molecular properties of an alpha-galactosidase synthesized and secreted by aspergillus nidulans.alpha-galactosidases from mycelial extract and culture filtrate of aspergillus nidulans have been purified to homogeneity and utilised to obtain polyclonal antibodies anti-alpha-galactosidase. the enzymatic characteristics and the cross reactivity of the antibodies suggest that alpha-galactosidases isolated from the two sources were the same enzyme. thus, a. nidulans synthesized and secreted only one enzymatic form of alpha-galactosidase which is a multimeric enzyme of 370 kda composed of four m ...19938405947
ph regulation is a major determinant in expression of a fungal penicillin biosynthetic gene.transcription of the ipna gene encoding isopenicillin n synthetase, an enzyme of secondary metabolism, is under the control of the ph regulatory system in the fungus aspergillus nidulans. external alkaline ph or mutations in pacc, the wide domain regulatory gene which mediates ph regulation, override carbon regulation of ipna transcript levels, resulting in elevation of the levels of this message in sucrose broth. strains carrying these mutations, which mimic growth at alkaline ph, produce highe ...19938404862
overproduction of, and interaction within, bifunctional domains from the amino- and carboxy-termini of the pentafunctional arom protein of aspergillus nidulans.the pentafunctional arom protein in aspergillus nidulans and other fungi catalyses five consecutive enzymatic steps leading to the production of 5-enolpyruvylshikimate 3-phosphate (epsp) in the shikimate pathway. the arom protein has five separate enzymatic domains that have previously been shown to display a range of abilities to fold and function in isolation as monofunctional enzymes. in this communication, we report (1) the stable overproduction of a bifunctional protein containing the 3-deh ...19938393515
the molecular biology of the pentafunctional arom protein. 19938383607
development of a transformation system for the filamentous, ml-236b (compactin)--producing fungus penicillium citrinum.we present here the first report of a transformation system developed for the filamentous, ml-236b (compactin)-producing fungus penicillium citrinum. hygromycin b-resistant colonies were obtained after treatment of protoplasts with a vector containing an escherichia coli hygromycin b phosphotransferase gene fused to a 3-phosphoglycerate kinase promoter from aspergillus nidulans. the transformation rate was 194 transformants per microgram circular dna per 4 x 10(5) viable protoplasts under optimi ...19938381335
quantitative structure-activity relationship models correctly predict the toxic and aneuploidizing properties of six halogenated methanes in aspergillus a previous study, the relationships between the chemical structure and the ability of 35 chlorinated aliphatic hydrocarbons to induce aneuploidy and toxicity in aspergillus nidulans were analyzed. quantitative structure-activity relationships (qsar) were defined for each of the biological activities under study: arr (the dose able to block mitotic growth), d37 (the dose with 37% of survival) and lec (the lowest efficient concentration in aneuploidy induction). in this study, these qsar equati ...19938377647
mutation in the bimd gene of aspergillus nidulans confers a conditional mitotic block and sensitivity to dna damaging agents.mutation in the bimd gene of aspergillus nidulans results in a mitotic block in anaphase characterized by a defective mitosis. mutation in bimd also confers, at temperatures permissive for the mitotic arrest phenotype, an increased sensitivity to dna damaging agents, including methyl methanesulfonate and ultraviolet light. in order to better understand the relationship between dna damage and mitotic progression, we cloned the bimd gene from aspergillus. a cosmid containing the bimd gene was iden ...19938375649
analysis of heterokaryon incompatibility between heterokaryon-compatibility (h-c) groups r and gl provides evidence that at least eight het loci control somatic incompatibility in aspergillus nidulans.protoplast fusion was used to hybridize strains 99-6 (h-cr) and 7-141 (h-cgl) in the birmingham collection of aspergillus nidulans. a sparsely conidiating, brown-pigmented diploid strain (ma7) was isolated. inocula from ma7 were haploidized on medium containing benlate and a haploid progeny sample collected. heterokaryon compatibility testing of selected progeny strains assayed each linkage group in turn and established that h-cr and h-cgl were hetero-allelic for heterokaryon incompatibility (he ...19938371120
the nia gene of fusarium oxysporum: isolation, sequence and development of a homologous transformation system.the fusarium oxysporum gene nia, encoding nitrate reductase (nr), was isolated from a cosmid library by direct complementation of an f. oxysporum nia- mutant. the gene specifies a protein of 905 amino acids and contains a 57-bp intron. comparison of the deduced aa sequence with nr of other fungi revealed a high degree of similarity and conservation in the intron position. the cloned nia made it possible to develop the first homologous transformation system for this fungus. transformation frequen ...19938370541
the gamma-tubulin gene of the malaria parasite plasmodium screening of cdna and genomic libraries of plasmodium falciparum with a dna probe derived from the cognate beta-tubulin gene, gene pf gamma tub has been identified that codes for gamma-tubulin, a newly discovered member of the tubulin superfamily that is indispensible for nuclear division and microtubule assembly [12]. gene pf gamma tub is not interrupted by introns and only present as a single-copy in the parasite genome. its encoded amino acid sequence (452 amino acids; m(r) 50,560) has a 6 ...19938366893
characterization of the pyruvate kinase-encoding gene (pki1) of trichoderma reesei.the pyruvate kinase-encoding gene (pki1) from trichoderma reesei was isolated by hybridization to the corresponding aspergillus nidulans pkia gene. the 1614-bp nucleotide (nt) sequence of the cloned gene codes for a 538-amino-acid protein. the coding sequence contains a single intron of 246 nt at a position identical to that of intron e in the a. nidulans gene. the pki protein shows extensive homology to the pkis of a. nidulans and a. niger (67%) and saccharomyces cerevisiae (59%). the 5' non-co ...19938359694
the aspergillus niger carbon catabolite repressor encoding gene, order to undertake a comparative analysis of carbon catabolite repression in two aspergillus species, the crea gene has been isolated from a. niger by cross hybridization, using the cloned a. nidulans gene. the a. niger gene has been shown to be functional in a. nidulans by heterologous complementation of the crea204 mutation of a. nidulans. overall, the genes show 90% sequence similarity (82% identity) at the amino acid (aa) level. there were some striking similarities between the aa sequenc ...19938359691
uvsi mutants defective in uv mutagenesis define a fourth epistatic group of uvs genes in aspergillus.three uv-sensitive mutations of a. nidulans, uvsi, uvsj and uvsa, were tested for epistatic relationships with members of the previously established groups, here called the "uvsf", "uvsc", and "uvsb" groups. uvsi mutants are defective for spontaneous and induced reversion of certain point mutations and differ also for other properties from previously analyzed uvs types. they are very sensitive to the killing effects of uv-light and 4-nqo (4-nitro-quinoline-n-oxide) but not to mms (methylmethane ...19938358834
either alpha-tubulin isogene product is sufficient for microtubule function during all stages of growth and differentiation in aspergillus nidulans.the filamentous fungus aspergillus nidulans has two genes encoding alpha-tubulin, tuba and tubb, which are differentially required at distinct stages during the life cycle. the tuba gene is required during vegetative growth for mitosis and nuclear migration (b. r. oakley, c. e. oakley, and j. e. rinehart, mol. gen. genet. 208:135-144, 1987; p. doshi, c. a. bossie, j. h. doonan, g. s. may, and n. r. morris, mol. gen. genet. 225:129-141, 1991). the tubb gene is not required for any detectable aspe ...19938336695
the isolation of mutagen-sensitive nuv mutants of aspergillus nidulans and their effects on mitotic recombination.more than 200 mutants of aspergillus nidulans were isolated as hypersensitive to the monofunctional alkylating agent mnng and/or uv-irradiation (designated nuv mutants). of these, 23 were selected for further characterization. all were markedly hypersensitive to both mnng and the quasi-uv-mimetic mutagen 4-nqo. the hypersensitive phenotype of each mutant was shown to result from mutation of a single gene. the nuv mutants exhibited a diverse range of growth responses on solid media containing var ...19938325481
complement binding to aspergillus conidia correlates with pathogenicity.complement has significant effects on the phagocytosis of aspergillus organisms. we examined the amount and type of complement component c3 bound to the resting conidia of 29 isolates from nine aspergillus species. the highly pathogenic species a. fumigatus and a. flavus bound fewer c3 molecules per unit of conidial surface area than did the less pathogenic species a. glaucus, a. nidulans, a. niger, a. ochraceus, a. terreus, a. versicolor, and a. wentii, as determined by quantitative flow cytome ...19938320488
isolation and nucleotide sequence of the 5-aminolevulinate synthase gene from aspergillus nidulans.the structural gene for 5-aminolevulinate (ala) synthase has been cloned and sequenced from the filamentous fungus aspergillus nidulans using an oligonucleotide probe based on a highly conserved-amino-acid sequence found in ala synthase genes of a wide range of species. the cloned gene, hema, has a 5' untranslated mrna of 92 nucleotides (nt) and one intron (64 nt). the deduced protein sequence (648 amino acids) shows 64% identity to the yeast ala synthase in the c-terminal region of 453 amino ac ...19938319309
choline: its role in the growth of filamentous fungi and the regulation of mycelial morphology.choline is an essential metabolite for the growth of filamentous fungi. it occurs most notably as a component of the major membrane phospholipid, phosphatidyl choline (lecithin), and fulfills a major role in sulphate metabolism in the form of choline-o-sulphate in many species. choline is usually synthesised endogenously, but exogenous choline can also be taken up, either to compensate for metabolic deficiencies in choline-requiring mutants such as those of aspergillus nidulans and neurospora cr ...19938318261
spatial and temporal controls of the aspergillus brla developmental regulatory gene.the aspergillus nidulans brla gene encodes a transcriptional regulator of central importance in controlling conidiophore development. i have determined the effects of mutations in other developmental regulatory genes on expression of a brla-lacz fusion gene. deletion of brla reduced beta-galactosidase levels by half and led to delocalization of enzyme accumulation. the meda26 and abaa2 developmental mutations led to overexpression of the fusion gene without altering spatial specificity. in contr ...19938316075
evidence for a multi-allelic heterokaryon incompatibility (het) locus detected by hybridization among three heterokaryon-compatibility (h-c) groups of aspergillus nidulans.a strain of heterokaryon-compatibility (h-c) group a was crossed sexually to strains of h-cb and h-cgl of aspergillus nidulans. a back-crossing programme established that there were seven hetero-allelic heterokaryon compatibility (het) genes controlling somatic incompatibility between strains of h-ca and h-cb. a similar back-crossing programme between strains of h-ca and h-cgl confirmed that these two groups differ at six het loci. previous work has shown that h-cb differs from h-cgl at two het ...19938314715
two different, adjacent and divergent zinc finger binding sites are necessary for crea-mediated carbon catabolite repression in the proline gene cluster of aspergillus nidulans.crea is the negative regulator mediating carbon catabolism repression in aspergillus nidulans. we have determined all the sites in the dna region between the prnd and prnb genes of the proline degradation cluster of this organism which are able to bind a fusion protein containing the zinc finger domain of crea. the consensus sequence derived for crea binding is 5'-syggrg-3', but not all possible sites derived from this consensus do in fact bind. the binding of at least some sequences of the form ...19948313886
cloning and characterization of the abfb gene coding for the major alpha-l-arabinofuranosidase (abf b) of aspergillus niger.based on amino-acid sequence data from aspergillus niger alpha-l-arabinofuranosidase b (abf b), and cyanogen bromide fragments derived thereof, deoxyoligonucleotide mixtures were designed to be employed as primers in a polymerase chain reaction (pcr) on a. niger genomic dna. this resulted in amplification of three related pcr products. the abfb gene encoding abf b was isolated from a genomic library using such an amplification product as a probe. a 5.1-kb bamhi fragment was subcloned to result i ...19938299175
co-transformation with autonomously-replicating helper plasmids facilitates gene cloning from an aspergillus nidulans gene library.autonomously-replicating, marker-less "helper" plasmids were added to transformations of aspergillus nidulans with plasmids which normally transform by chromosomal integration. this resulted in as much as a 200-fold increase in transformation efficiency. recovery of autonomously-replicating plasmid co-integrates indicated that co-transformation involves recombination between integrating and helper plasmids, which occurs at a high frequency. increasing dna sequence-homology between pairs of plasm ...19938299174
induction of glucose oxidase, catalase, and lactonase in aspergillus niger.the induction of glucose oxidase, catalase, and lactonase activities was studied both in wild-type and in glucose oxidase regulatory and structural mutants of aspergillus niger. the structural gene for glucose oxidase was isolated and used for northern analysis and in transformation experiments using various gox mutations. wild-type phenotype could be restored in the glucose oxidase-negative mutant (goxc) by transformation with the structural gene. we conclude, therefore, that the goxc marker wh ...19938299156
molecular organization of the escherichia coli gab cluster: nucleotide sequence of the structural genes gabd and gabp and expression of the gaba permease gene.we have determined the nucleotide sequences of two structural genes of the escherichia coli gab cluster, which encodes the enzymes of the 4-aminobutyrate degradation pathway: gabd, coding for succinic semialdehyde dehydrogenase (ssdh, ec and gabp, coding for the 4-aminobutyrate (gaba) transport carrier (gaba permease). we have previously reported the nucleotide sequence of the third structural gene of the cluster, gabt, coding for glutamate: succinic semialdehyde transaminase (ec 2.6.1 ...19938297211
genesis of eukaryotic transcriptional activator and repressor proteins by splitting a multidomain anabolic enzyme.the genes necessary for the correctly regulated catabolism of quinate in aspergillus nidulans and neurospora crassa are controlled at the level of transcription by a dna-binding activator protein and a repressor protein that directly interact with one another. the repressor protein is homologous throughout its length with the three c-terminal domains of a pentafunctional enzyme catalysing five consecutive steps in the related anabolic shikimate pathway. we now report that the activator protein i ...19938294040
the macronuclear gamma-tubulin-encoding gene of euplotes octocarinatus contains two introns and an in-frame tga.the gamma-tubulin (gamma-tub)-encoding gene (gamma-tub) of euplotes octocarinatus was amplified from macronuclear dna with the help of the polymerase chain reaction (pcr) and sequenced. the polypeptide deduced from the gene consists of 462 amino acids (aa). it shares 61% aa identity with the aspergillus nidulans gamma-tub. the gene contains an in-frame tga codon and two small pre-mrna introns (36 and 26 bp). we suggest that the tga, like tga codons in the pheromone-encoding genes of e. octocarin ...19938294024
analysis of the regulation of penicillin biosynthesis in aspergillus nidulans by targeted disruption of the acva analyse the regulation of the biosynthesis of the secondary metabolite penicillin in aspergillus nidulans, a strain with an inactivated acva gene produced by targeted disruption was used. acva encodes delta-(l-alpha-aminoadipyl)-l-cysteinyl-d-valine synthetase (acvs), which catalyses the first step in the penicillin biosynthetic pathway. to study the effect of the inactivated acva gene on the expression of acva and the second gene, ipna, which encodes isopenicillin n synthase (ipns), a. nidul ...19948277946
relationship between zinc content and dna-binding activity of the dna-binding motif of the transcription factor alcr in aspergillus nidulans.the transcription factor alcr of the ethanol utilisation pathway in aspergillus nidulans contains a zinc binuclear motif (cysx2cysx6cysx16cysx2cysx6cys), within the dna-binding domain located in the n-terminal region of the alcr protein. specific targets have been localised in the promoter of the alcr gene, involved in the autoregulation process, and in the promoter of the structural gene alca (encoding alcohol dehydrogenase i), which is also under the control of alcr. the dna-binding domain has ...19948277945
cloning and characterisation of the cytochrome c gene of aspergillus nidulans.the cytochrome c gene (cyca) of the filamentous fungus aspergillus nidulans has been isolated and sequenced. the gene is present in a single copy per haploid genome and encodes a polypeptide of 112 amino acid residues. the nucleotide sequence of the a. nidulans cyca gene shows 87% identity to the dna sequence of the neurospora crassa cytochrome c gene, and approximately 72% identity to the sequence of the saccharomyces cerevisiae iso-1-cytochrome c gene (cyc1). the s. cerevisiae cyc1 gene was us ...19948277943
purification and characterization of glucose-6-phosphate dehydrogenase from aspergillus niger and aspergillus nidulans.glucose-6-phosphate dehydrogenase (g6pd; d-glucose 6-phosphate:nadp+ oxidoreductase, ec has been purified from aspergillus nidulans and aspergillus niger by a combination of affinity and anion exchange chromatography. a 500-1000-fold purification was obtained and the final enzyme preparations were shown to be pure but not homogeneous. for both fungi the purified enzyme preparation gave two bands on native and denaturing gels. the catalytically active form is a multimer. the molecular m ...19938277259
identification of 21 novel human protein kinases, including 3 members of a family related to the cell cycle regulator nima of aspergillus nidulans.the nima gene encodes a protein-serine/threonine kinase that is required along with the p34cdc2 kinase for mitosis in aspergillus nidulans. we have searched for human protein kinases that are related to the nima protein kinase using the polymerase chain reaction. different pairs of degenerate oligonucleotides specific for conserved amino acid motifs in the catalytic domain of nima were used as primers in the polymerase chain reaction to amplify partial complementary dnas (cdnas) of protein kinas ...19938274451
survey of mycoflora and mycotoxins in egyptian soybean seeds.after four months in commercial storage, 100 soybean samples from different places of egyptian governorates were assayed for filamentous fungal growth at two incubation temperatures (28 and 45 degrees c). 73 species and 8 varieties belonging to 32 genera were isolated by the dilution plate method. at 28 degrees c, the common species were aspergillus flavus, a. fumigatus, a. niger and a. alutaceus, followed by a. terreus, penicillium chrysogenum, p. citrinum, mucor hiemalis, m. racemosus, emerice ...19938271157
kinetic studies of nitrite uptake by aspergillus nidulans.1. in the filamentous mold aspergillus nidulans, net nitrite uptake is inducible by nitrate and nitrite, is probably different from the nitrate uptake system and is partially repressed by ammonium. 2. the concentration dependence of net nitrite uptake by the bia1 faca303 strain of a. nidulans shows a saturation kinetics with an apparent km value of 0.64 mm. 3. strains of a. nidulans carrying the niha1 and niha1 chla14 mutations considerably reduced the affinity of the nitrite uptake system for t ...19938257915
characterization of the 3-dehydroquinase domain of the pentafunctional arom protein, and the quinate dehydrogenase from aspergillus nidulans, and the overproduction of the type ii 3-dehydroquinase from neurospora crassa.the arom protein of aspergillus nidulans is a multidomain pentafunctional polypeptide that is active as a dimer and catalyses steps 2-6 in the prechorismate section of the shikimate pathway. the three c-terminal domains (including the type i 3-dehydroquinase) of the arom protein are homologous with the qutr-encoded qutr protein that represses transcription of the eight genes comprising the quinic acid utilization (qut) gene cluster, and the two n-terminal domains are homologous with the quta-enc ...19938257437
regulation of sulfur and nitrogen metabolism in filamentous the filamentous fungi, n. crassa and a. nidulans, complex regulatory circuits control nitrogen metabolism and sulfur metabolism. the expression of entire sets of unlinked structural genes that encode metabolic enzymes is repressed when favored sulfur or nitrogen sources are available. these structural genes are coregulated by global positive-acting regulatory proteins and often are also controlled by metabolic inducers and pathway-specific regulatory proteins. the recent isolation of regulato ...19938257101
comparative studies on o-acetylhomoserine sulfhydrylase: physiological role and characterization of the aspergillus nidulans enzyme.o-acetylhomoserine sulfhydrylase (oah shlase) from aspergillus nidulans is an oligomeric protein with a broad substrate specificity with regard to sulfhydryl compounds. as its saccharomyces cerevisiae counterpart the enzyme also reacts with o-acetylserine and is inhibited by carbonyl reagents but not by antiserum raised against the yeast enzyme. in contrast to saccharomyces cerevisiae the enzyme is not essential for aspergillus nidulans as indicated by the completely prototrophic phenotype of oa ...19938249501
arabinan degrading enzymes from aspergillus nidulans: induction and purification.the presence in aspergillus nidulans of two enzymes related to the aspergillus niger endo-arabinase and alpha-l-arabinofuranosidase b has been established using antibodies against the purified a. niger enzymes. moreover, the absence of an equivalent in a. nidulans to the alpha-l-arabinofuranosidase a of a. niger has been confirmed both at the protein and at the dna level. both a. nidulans arabinases have been purified and physico-chemically and kinetically characterized. they have a much higher ...19938243977
brushing up on bristles: complex genes and morphogenesis in molds. 19938236455
molecular characterization of a fungal secondary metabolism promoter: transcription of the aspergillus nidulans isopenicillin n synthetase gene is modulated by upstream negative elements.the aspergillus nidulans ipns gene, encoding isopenicillin n synthetase, is a secondary metabolism gene. it is contiguous to, but divergently transcribed from, the acvs gene at the penicillin gene cluster. the untranslated region between both orfs is 872bp long. here we present the physical and functional characterization of the ipns transcriptional unit. transcriptional start point (tsp) mapping reveals heterogeneity at the 5'-end of the mrna, with a major start at -106 relative to the initiati ...19938231816
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