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  • 1
    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.
    Type of Medium: Electronic Resource
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  • 2
    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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  • 3
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    International Journal for Numerical Methods in Fluids 24 (1997), S. 875-892 
    ISSN: 0271-2091
    Keywords: adaptive parallel multigrid method ; local refinement criteria ; incompressible Navier-Stokes equations ; hole pressure problem ; Engineering ; Numerical Methods and Modeling
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: In this paper an adaptive parallel multigrid method and an application example for the 2D incompressible Navier-Stokes equations are described. The strategy of the adaptivity in the sense of local grid refinement in the multigrid context is the multilevel adaptive technique (MLAT) suggested by Brandt. The parallelization of this method on scalable parallel systems is based on the portable communication library CLIC and the message-passing standards: PARMACS, PVM and MPI. The specific problem considered in this work is a two-dimensional hole pressure problem in which a Poiseuille channel flow is disturbed by a cavity on one side of the channel. Near geometric singularities a very fine grid is needed for obtaining an accurate solution of the pressure value. Two important issues of the efficiency of adaptive parallel multigrid algorithms, namely the data redistribution strategy and the refinement criterion, are discussed here. For approximate dynamic load balancing, new data in the adaptive steps are redistributed into distributed memories in different processors of the parallel system by block remapping. Among several refinement criteria tested in this work, the most suitable one for the specific problem is that based on finite-element residuals from the point of view of self-adaptivity and computational efficiency, since it is a kind of error indicator and can stop refinement algorithms in a natural way for a given tolerance. Comparisons between different global grids without and with local refinement have shown the advantages of the self-adaptive technique, as this can save computer memory and speed up the computing time several times without impairing the numerical accuracy. © 1997 By John Wiley & Sons, Ltd. Int. J. Numer. Methods Fluids 24, 875-892, 1997.
    Additional Material: 13 Ill.
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  • 4
    ISSN: 0935-9648
    Keywords: Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics , Physics
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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