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  • 1
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Many yeast species can utilize glycerol, both as a sole carbon source and as an osmolyte. In Saccharomyces cerevisiae, physiological studies have previously shown the presence of an active uptake system driven by electrogenic proton symport. We have used transposon mutagenesis to isolate mutants affected in the transport of glycerol into the cell. Here we present the identification of YGL084c, encoding a multimembrane-spanning protein, as being essential for proton symport of glycerol into S. cerevisiae. The gene is named GUP1 (glycerol uptake) and, for growth on glycerol, is important as a carbon and energy source. In addition, in strains deficient in glycerol production it also provides osmotic protection by the addition of glycerol. Another open reading frame (ORF), YPL189w, presenting a high degree of homology to YGL084c, similarly appears to be involved in active glycerol uptake in salt-containing glucose-based media in strains deficient in glycerol production. Analogously, this gene is named GUP2. To our knowledge, this is the first report on a gene product involved in active transport of glycerol in yeasts. Mutations with the same phenotypes occurred in two other ORFs of previously unknown function, YDL074c and YPL180w.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 142 (1996), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Pichia sorbitophila, a yeast species that is highly resistant to osmotic stress in general and to salt stress in particular, was subjected to a mutagenesis strategy in order to obtain mutants deficient in the glycerol active uptake previously described. Density centrifugation was used for enrichment of NaCl sensitive mutants in either glucose or glycerol media. Several phenotypic classes of mutants were identified, to which physiological tests were applied concerning the activity of the symporter, its accumulation capacity and the detection of the activity of glycerol pathway specific enzymes. From these, two mutant strains were selected, presenting a clearly deficient phenotype on H+/glycerol symport activity.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 11 (1995), S. 111-119 
    ISSN: 0749-503X
    Keywords: Pichia sorbitophila ; halotolerance ; osmoregulation ; glycerol transport ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Pichia sorbitophila is a halotolerant yeast capable of surviving to extracellular NaCl concentrations up to 4 M in mineral medium when glucose or glycerol are the only carbon and energy sources. Evidence is presented here that glycerol, the main compatible solute this yeast accumulates so as to maintain osmotic balance, is actively co-transported with protons. This transport system was shown to be constitutive, not needing induction by either glycerol or salt, and was not repressible by glucose. In glucose- or glycerol-grown cells, a simple diffusion was detectable, and iterative calculations were performed to calculate kinetic parameters, in the presence and in the absence of NaCl. At 25°C, pH 5·0, in glucose-grown cells these were: Km = 0·81 ± 0·11 mM and Vmax = 634·2 ± 164·8 μmol h-1 per g (glycerol); Km = 1·28 ± 0·60 mM and Vmax = 558·6 · 100·6 μmol h-1 per g (protons). Correspondent stoichiometry was approximately 1, either for these conditions or in the presence of 1 M-NaCl. An increase in acumulation capacity was evident when different concentrations of NaCl were present. This capacity was shown to be dependent on ΔpH and membrane potential, consistently with an electrogenic character. We suggest that the main role of this system is in osmoregulation, by keeping glycerol accumulated inside the cells, compensating for leakage, due to its liposoluble character.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 187-191 
    ISSN: 0749-503X
    Keywords: Halotolerance ; osmoregulation ; glycerol-sodium symport ; glycerol-potassium symport ; sodium-proton exchange ; Debaryomyces hansenii ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Several authors have shown that the halotolerant yeast Debaryomyces hansenii, when growing exponentially in glucose medium in the presence of sodium chloride, maintains osmotic balance by establishing sodium and glycerol gradients of opposite signs across the plasma membrane. Evidence is presented here that the two gradients are linked through a sodium-glycerol symport that uses the sodium gradient as a driving force for maintaining the glycerol gradient. The symporter also accepts potassium ions as co-substrate. The kinetic parameters at 25°C, pH 5·0 were the following: Vmax, decreasing from over 500 to less than 40 μmol g-1 per h over a concentration range of 0-3 M extracellular sodium chloride; Km (glycerol) 0·40-0·6 mM over the same range; Km (sodium ions) 16·0 ± 3·21μM; Km, (potassium ions) 10·4 ± 3·6μM. Furthermore, it was observed that glycerol uptake was accompanied by proton uptake when extracellular sodium chloride was present and that the protonophore carbonylcyanide-M-chlorophenylhydrazone induced collapse of the glycerol gradient, supporting earlier proposals by others that the sodium gradient is maintained by an active sodium-proton exchange mechanism.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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