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  • Chemistry  (19)
  • Saccharomyces cerevisiae  (6)
  • Malate synthase
  • Earth model, also for more shallow analyses !
  • Lactobacillus delbrueckii subsp. lactis
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  • 1
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Fructose-1,6-bisphosphatase ; Glucose repression ; Gene activation ; Gluconeogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Fructose-1,6-bisphosphatase is a key enzyme in gluconeogenesis and the FBP1 gene is not transcribed during growth with glucose. Genetic analysis indicated a positive regulation of FBP1 expression after exhaustion of glucose. By linker-deletion analysis, two upstream activation sites (UAS1 and UAS2) were localized and the respective UAS-binding factors (DAP I and DAP II for derepression activating protein) were identified by gel retardation. UAS1 and UAS2 span about 30 bp each, and are separated by approximately 30 bp. Both UAS sites act synergistically. Although UAS1 showed some similarities to the DNA-binding consensus for the general yeast activator Rap1, competition experiments and DEAE-chromatography proved that DAP I and Rap1 correspond to different proteins. Gel retardation by DAP I depended on carbon sources and did not occur in cells growing logarithmically with glucose, whereas a strong retardation signal was obtained with ethanol-grown cells. The present results suggest that DAP I and DAP II are the final regulatory elements for glucose derepression.
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  • 2
    ISSN: 1432-0983
    Keywords: Key words Transcriptional regulation ; Phospholipid biosynthesis ; Saccharomyces cerevisiae ; INO2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Expression of structural genes of phospholipid biosynthesis in yeast is mediated by the inositol/choline-responsive element (ICRE). ICRE-dependent gene activation, requiring the regulatory genes INO2 and INO4, is repressed in the presence of the phospholipid precursors inositol and choline. INO2 and, to a less extent, INO4 are positively autoregulated by functional ICRE sequences in the respective upstream regions. However, an INO2 allele devoid of its ICRE functionally complemented an ino2 mutation and completely restored inositol/choline regulation of Ino2p-dependent reporter genes. Low-level expression of INO2 and INO4 genes, each under control of the heterologous MET25 promoter, did not alter the regulatory pattern of target genes. Thus, upstream regions of INO2 and INO4 are not crucial for transcriptional control of ICRE-dependent genes by inositol and choline. Interestingly, over-expression of INO2, but not of INO4, counteracted repression by phospholipid precursors. Possibly, a functional antagonism between INO2 and a negative regulator is the key event responsible for repression or de-repression.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 23 (1993), S. 375-381 
    ISSN: 1432-0983
    Keywords: Saccharomyces cerevisiae ; Isocitrate lyase ; Gene regulation ; Ethanol induction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ICL1 gene encoding the isocitrate lyase from Saccharomyces cerevisiae was cloned and sequenced. A reading frame of 557 amino acids showing significant similarity to isocitrate lyases from seven other species could be identified. Construction of icl1 null mutants led to growth defects on C2 carbon sources while utilization of sugars or C3 substrates remained unaffected. Using an ICL1-lacZ fusion integrated at the ICL1 locus, a more than 200-fold induction of β-galactosidase activity was observed after growth on ethanol when compared with glucose-repressed conditions. A preliminary analysis of the ICL1 upstream region identified a 364-bp fragment necessary and sufficient for this regulatory phenotype. Sequence motifs also present in the upstream regions of co-regulated genes were found within this region.
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  • 4
    ISSN: 1617-4623
    Keywords: Key wordsSaccharomyces cerevisiae ; Gluconeogenesis ; Malate synthase ; Transcriptional regulation ; MLS1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The malate synthase gene, MLS1, of the yeast Saccharomyces cerevisiae is transcriptionally regulated by the carbon source in the growth medium. A MLS1-lacZ fusion gene, expressed at a basal level in the presence of 2% glucose, is derepressed more than 100-fold under conditions of sugar limitation. No evidence for MLS1 induction by oleic acid was found. By deletion analysis of the MLS1 control region, we identified two sites, UAS1 and UAS2, as important for efficient derepression of the gene. Both sites contain sequences that resemble the previously characterized carbon source-responsive element (CSRE) found in the promoter of the isocitrate lyase gene ICL1. Indeed, UAS1 and UAS2 in the MLS1 upstream region turn out to be functional CSRE sequence variants. This finding allowed us to define a modified version of the CSRE consensus sequence (CCRTYSRNCCG). Protein binding to UAS1MLS1 was observed with extracts from derepressed but not from repressed cells, and could be competed for by an excess of the unlabelled CSRE(ICL1) sequence. No competition was observed with a mutated CSRE variant. Site-directed mutagenesis of both CSREs in the MLS1 promoter reduced gene activation under derepressing conditions to 20% of the wild-type level. The same decrease was observed with the wild-type MLS1 promoter in a cat8 mutant, lacking an activator of CSRE-dependent transcription. The CSRE/Cat8p-independent activation of MLS1 is mediated by constitutive UAS elements. The pleiotropic transcription factor Abf1p, which binds to the MLS1 upstream region, may contribute to constitutive activation. Thus, in order to ensure the severe glucose repression of MLS1 observed, repressor elements that respond to the carbon source must counteract constitutive activation. In summary, ICL1 and MLS1 share common cis-acting elements, although a distinct mechanism of carbon source control also contributes to MLS1 regulation.
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  • 5
    ISSN: 1617-4623
    Keywords: GC clusters ; Mobile elements ; Target sites ; mtDNA ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary GC clusters constitute the major repetitive elements in the mitochondrial (mt) genome of the yeast Saccharomyces cerevisiae. Many of these clusters are optional and thus contribute much to the polymorphism of yeast mtDNAs. We have made a systematic search for polymorphic sites by comparing mtDNA sequences of various yeast strains. Most of the 26 di- or polymorphic sites found differ by the presence or absence of a GC cluster of the majority class, here referred to as the M class, which terminate with an AGGAG motif. Comparison of sequences with and without the GC clusters reveal that elements of the subclasses M1 and M2 are inserted 3′ to a TAG, flanked by A+T rich sequences. M3 elements, in contrast, only occur in tandem arrays of two to four GC clusters; they are consistently inserted 3′ to the AGGAG terminal sequence of a preexisting cluster. The TAG or the terminal AGGAG, therefore, are regarded as being part of the target sites for M1 and M2 or M3 elements, respectively. The dinucleotide AG is in common to both target sites; it also occurs at the 3′ terminus (AGGAG). This suggests its duplication during GC cluster insertion. This notion is supported by the observation that GC clusters of the minor classes G and V similarily repeat at their 3′ terminus a GT or an AA dinucleotide, respectively, from their putative target sites.
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  • 6
    ISSN: 1617-4623
    Keywords: Glucose repression ; Glucose derepression ; Regulatory genes ; Expression analysis ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Yeast strains carrying one of the two regulatory mutations cat1 and cat3 are defectve in derepression of several glucose-repressible enzymes that are necessary for utilizing non-fermentable carbon sources. Hence, these strains fail to grow on ethanol, glycerol or acetate. The synthesis of isocitrate lyase, malate synthase, malate dehydrogenase and fructose-1,6-bisphosphatase is strongly affected in cat1 and cat3 strains. Genes CAT1 and CAT3 have been isolated by complementation of the cognate, mutations after transformation with an episomal plasmid gene library. The restriction map of CAT1 proved its allelism to the earlier isolated SNF1 gene. Both genes appear to exist as single-copy genes per haploid genome as indicated by Southern hybridization. Northern analysis has shown that the 1.35 kb CAT3 mRNA is constitutively expressed, independent of the carbon source in the medium. Derepression studies with CAT3 transformants using a multi-copy plasmid showed over-expression of glyoxylate cycle enzymes. This result would be consistent with a direct effector function for the CAT3 gene product.
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  • 7
    ISSN: 1617-4623
    Keywords: Prolidase ; Metalloprotease ; Lactobacillus delbrueckii subsp. lactis ; Nucleotide sequence analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract From a genomic library of Lactobacillus delbrueckii subsp. lactis (DSM7290) DNA, in the low-copy-number vector pLG339, a recombinant clone was selected, which complemented a mutation in the prolidase gene (pepQ) of Escherichia coli UK173. Nucleotide sequence analysis revealed an open reading frame of 1104 nucleotides corresponding to a protein of 368 amino acids with a calculated pI of 4.64 and a molecular mass of 41087 Da. The start site of pepQ transcription was determined by primer extension analysis with mRNA prepared from L. delbrueckii. Based on homology of the gene product to various peptidases and on the substrate specificity determined, the peptidase was designated PepQ. The influence of various protease inhibitors and cations on peptidase activity indicated that PepQ is a metalloprotease. The absence of a membrane-spanning domain and a signal peptide sequence argues for a cytoplasmic localization of the enzyme.
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  • 8
    Electronic Resource
    Electronic Resource
    Weinheim : Wiley-Blackwell
    Zeitschrift für anorganische Chemie 40 (1904), S. 385-399 
    ISSN: 0863-1778
    Keywords: Chemistry ; Inorganic Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Additional Material: 3 Ill.
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  • 9
    ISSN: 1052-9306
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: A specific and sensitive gas chromatographic/mass spectrometric method was developed and validated for the determination of the antidepressant levoprotiline in blood, plasma and urine and the simultaneous determination of levoprotiline and its desmethyl metabolite in urine. Deuterium-labelled analogues were used as internal standards. The compounds were isolated from the biological fluids by liquid-liquid extraction under basic conditions. Following derivatization with perfluoropropionic anhydride, the samples were analysed by capillary column gas chroma-tography/electron impact mass spectrometry with selected ion monitoring. The analysis of spiked samples demonstrated the high accuracy and precision of the method. Blood concentrations of levoprotiline down to 0.7 nmol I-1 (1 ml used for analysis) could be quantified with a coefficient of variation of 10% or less. The method is suitable for use in pharmacokinetic and bioavailability studies of levoprotiline in humans.
    Additional Material: 6 Ill.
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  • 10
    Electronic Resource
    Electronic Resource
    Weinheim [u.a.] : Wiley-Blackwell
    Materials and Corrosion/Werkstoffe und Korrosion 15 (1964), S. 621-631 
    ISSN: 0947-5117
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Description / Table of Contents: The relationship of the intercrystalline corrosion of a non-stabilised 18/10-chromium-nickel steel with some test solutions and with the potential in boiling 2 n H2SO4Specimens of an austenitic 18/20 nickelchromium steel with about 0.06 pC C, quenched at 1300° C and annealed for different periods of time at 550 or 675° C were potentiostatically subjected to the attack of boiling 2n-H2SO4, of a nitric / hydrofluoric acid pickle at room temperature, and of boiling acids in accordance with the Strauss, Huey and Streicher test. Subsequently, the corrosion rates were determined by weighing and the corrosion examined by visual inspection and microscopically.In the active and passive condition grain boundary corrosion is only encountered after sensitizing annealing. In this connection, the potential merely determines the intensity of the grain boundary attack, but not its occurrence as such. The influence of the potential is more marked at higher than at lower sensitizing annealing temperatures. The test results can easily be interpreted with the aid of the chrome reduction theory.In the transition zone to the transpassive condition, and in the latter, an intercrystalline corrosion attack occurs which depends but little on the sensitizing annealing, and which also occurs with solution-annealed specimens. It is also in this zone of the potential that the non-potentiostatic tests according to Huey and Streicher are carried out.The test results show a specific reaction to chrome reduction of the corrosion attack in the active and passive condition, i.e. the attack in the hydrofluoric/nitric acid pickle and in the Strauss solution. In contrast, the Huey and Streicher tests additionally bring in other properties which, in the main, have an influence on the excess potential of the cathodic part-reaction. These testing agents cannot be regarded as specific criteria for the grain decomposition proneness.
    Notes: Proben eines austenitischen 18/10-Chrom-Nickel-Stahles mit etwa 0,06% C, die von 1300° C abgeschreckt und verschieden lange bei 550 und 675° C geglüht worden waren, wurden potentiostatisch dem Angriff siedender 2n-H2SO4, einer Salpetersäure-Flußsäure-Beize bei Raumtemperatur sowie dem Angriff siedender Säuren entsprechend dem Strauß, Huey- und Streicher-Test unterworfen. Anschließend werden die Korrosionsgeschwindigkeiten durch Wägung bestimmt und der Korrosionsangriff visuell und mikroskopisch im Schliffbild untersucht.Im Aktiv- und Passivzustand tritt Korngrenzenkorrosion nur nach einer Sensibilisierungsglühbehandlung auf. Das Potential bestimmt hierbei nur die Intensität des Korngrenzenangriffs und nicht das Auftreten desselben an sich. Der Einfluß des Potentials ist bei hohen Sensibilisierungslühtemperaturen stärker ausgeprägt, als bei niedrigen Temperaturen. Die Versuchsergebnisse lassen sich mit Hilfe der Chromverarmungstheorie zwanglos deuten.Im Übergangsbereich zum Transpassivzustand erfolgt ein interkristalliner Angriff, der nur wenig von der Sensibilisierungsglühbehandlung abhängt und auch bei lösungsgeglühten Proben auftritt. In diesem Potentialbereich erfolgen auch die nicht potentiostatischen Prüfungen nach Huey und Streicher.Die Versuchsergebnisse zeigen, daß der Angriff im Aktiv- und Passivzustand, also der Angriff in der Flußsäure-Salpeter- Beize und in der Straußschen Lösung, spezifisch auf Chromverarmung ansprechen. Beim Huey- und Streicher-Test werden dagegen zusätzlich noch Eigenschaften, die im wesentlichen die Überspannung der kathodischen Teilreaktion beeinflussen, miterfaßt. Diese Prüfmittel können nicht als spezifisch für die Kornzerfallsanfälligkeit angesehen werden.
    Additional Material: 9 Ill.
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