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  • 1
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The RAS proteins of the yeast Saccharomyces cerevisiae fulfil a similar control function on yeast adenylate cyclase as the mammalian Gs proteins on mammalian adenylate cyclase. The discovery that glucose and other fermentable sugars act as specific activators of the RAS-adenylate cyclase pathway in yeast appeared to offer a mechanism for the way in which at least one nutrient would control progression over the start point in the G1 phase of the yeast cell cycle by means of this pathway. Recently, however, evidence has been obtained to show that the glucose-activation pathway of adenylate cyclase is a glucose-repressible pathway and therefore not operative during growth on glucose. In addition, mutant strains were obtained which lack the glucose-activation pathway and show normal exponential growth on glucose. This appears to confine the physiological role of this pathway to control of the transition from the derepressed state (growth on respirative carbon sources) to the repressed state (growth on fermentative carbon sources) by means of an already well-documented cAMP-triggered protein phosphorylation cascade. Intracellular acidification also stimulates the RAS-adenylate cyclase pathway, which might constitute a rescue mechanism for cells suffering from stress conditions. The presence of a nitrogen source does not stimulate the RAS-adenylate cyclase pathway. Although other nutrient signals for the pathway might still be discovered, it appears more and more likely that the well-known requirement of cAMP for progression over the start point of the yeast cell cycle is limited to providing a basal cAMP level rather than acting as a second messenger for an extracellular signal. A model In which a nitrogen-source-induced pathway leads to activation of cAMP-dependent protein kinase synergistically with cAMP, or alternatively causes activation only of the free catalytic subunits, would provide an elegant explanation for the apparent contradiction that nitrogen-source induced progression over the start point of the yeast cell cycle does not seem to be mediated by cAMP but on the other hand seems to depend on cAMP and cAMP-dependent protein kinase.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Anaerobic xylulose fermentation was compared in strains of Zygosaccharomyces and Saccharomyces cerevisiae, mutants and wild-type strains to identify host-strain background and genetic modifications beneficial to xylose fermentation. Overexpression of the gene (XKS1) for the pentose phosphate pathway (PPP) enzyme xylulokinase (XK) increased the ethanol yield by almost 85% and resulted in ethanol yields [0.61 C-mmol (C-mmol consumed xylulose)−1] that were close to the theoretical yield [0.67 C-mmol (C-mmol consumed xylulose)−1]. Likewise, deletion of gluconate 6-phosphate dehydrogenase (gnd1Δ) in the PPP and deletion of trehalose 6-phosphate synthase (tps1Δ) together with trehalose 6-phosphate phosphatase (tps2Δ) increased the ethanol yield by 30% and 20%, respectively. Strains deleted in the promoter of the phosphoglucose isomerase gene (PGI1) – resulting in reduced enzyme activities – increased the ethanol yield by 15%. Deletion of ribulose 5-phosphate (rpe1Δ) in the PPP abolished ethanol formation completely. Among non-transformed and parental strains S. cerevisiae ENY. WA-1A exhibited the highest ethanol yield, 0.47 C-mmol (C-mmol consumed xylulose)−1. Other non-transformed strains produced mainly arabinitol or xylitol from xylulose under anaerobic conditions. Contrary to previous reports S. cerevisiae T23D and CBS 8066 were not isogenic with respect to pentose metabolism. Whereas, CBS 8066 has been reported to have a high ethanol yield on xylulose, 0.46 C-mmol (C-mmol consumed xylulose)−1 (Yu et al. 1995), T23D only formed ethanol with a yield of 0.24 C-mmol (C-mmol consumed xylulose)−1. Strains producing arabinitol did not produce xylitol and vice versa. However, overexpression of XKS1 shifted polyol formation from xylitol to arabinitol.
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  • 3
    ISSN: 1432-072X
    Keywords: Key words Trehalase ; Trehalose ; Kluyveromyces lactis ; Protein phosphorylation ; NTH1 ; Glucose induction ; Glucose transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We cloned the Kluyveromyces lactis KlNTH1 gene, which encodes neutral trehalase. It showed 65.2% and 68.5% identity at nucleotide and amino acid sequence level, respectively, with the Saccharomyces cerevisiae NTH1 gene. Multiple alignment of the predicted trehalase protein sequences from yeasts, bacteria, insects, and mammals revealed two major domains of conservation. Only the yeast trehalases displayed in an N-terminal extension two consensus sites for cAMP-dependent protein phosphorylation and a putative Ca2+-binding sequence. Gene disruption of the KlNTH1 gene abolished neutral trehalase activity and clearly revealed a trehalase activity with an acid pH optimum. It also resulted in a high constitutive trehalose level. Expression of the KlNTH1 gene in an S. cerevisiae nth1Δ mutant resulted in rapid activation of the heterologous trehalase upon addition of glucose to cells growing on a nonfermentable carbon source and upon addition of a nitrogen source to cells starved for nitrogen in a glucose-containing medium. In K. lactis, the same responses were observed except that rapid activation by glucose was observed only in early-exponential-phase cells. Inactivation of K. lactis neutral trehalase by alkaline phosphatase and activation by cAMP in cell extracts are consistent with control of the enzyme by cAMP-dependent protein phosphorylation.
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  • 4
    ISSN: 1617-4623
    Keywords: Key words Trehalose synthase complex ; Stress-responsive element (STRE) ; Stress response ; Nutrients ; Ras-cAMP pathway
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Saccharomyces cerevisiae cells show under suboptimal growth conditions a complex response that leads to the acquisition of tolerance to different types of environmental stress. This response is characterised by enhanced expression of a number of genes which contain so-called stress-responsive elements (STREs) in their promoters. In addition, the cells accumulate under suboptimal conditions the putative stress protectant trehalose. In this work, we have examined the expression of four genes encoding subunits of the trehalose synthase complex, GGS1/TPS1, TPS2, TPS3 and TSL1. We show that expression of these genes is coregulated under stress conditions. Like for many other genes containing STREs, expression of the trehalose synthase genes is also induced by heat and osmotic stress and by nutrient starvation, and negatively regulated by the Ras-cAMP pathway. However, during fermentative growth only TSL1 shows an expression pattern like that of the STRE-controlled genes CTT1 and SSA3, while expression of the three other trehalose synthase genes is only transiently downregulated. This difference in expression might be related to the known requirement of trehalose biosynthesis for the control of yeast glycolysis and hence for fermentative growth. We conclude that the mere presence in the promoter of (an) active STRE(s) does not necessarily imply complete coregulation of expression. Additional mechanisms appear to fine tune the activity of STREs in order to adapt the expression of the downstream genes to specific requirements.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1617-4623
    Keywords: Trehalose synthase complex ; Stress-responsive element (STRE) ; Stress response ; Nutrients ; Ras-cAMP pathway
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Saccharomyces cerevisiae cells show under suboptimal growth conditions a complex response that leads to the acquisition of tolerance to different types of environmental stress. This response is characterised by enhanced expression of a number of genes which contain so-called stress-responsive elements (STREs) in their promoters. In addition, the cells accumulate under suboptimal conditions the putative stress protectant trehalose. In this work, we have examined the expression of four genes encoding subunits of the trehalose synthase complex,GGS1/TPS1, TPS2, TPS3 andTSL1. We show that expression of these genes is coregulated under stress conditions. Like for many other genes containing STREs, expression of the trehalose synthase genes is also induced by heat and osmotic stress and by nutrient starvation, and negatively regulated by the Ras-cAMP pathway. However, during fermentative growth onlyTSL1 shows an expression pattern like that of the STRE-controlled genesCTT1 andSSA3, while expression of the three other trehalose synthase genes is only transiently down-regulated. This difference in expression might be related to the known requirement of trehalose biosynthesis for the control of yeast glycolysis and hence for fermentative growth. We conclude that the mere presence in the promoter of (an) active STRE(s) does not necessarily imply complete coregulation of expression. Additional mechanisms appear to fine tune the activity of STREs in order to adapt the expression of the downstream genes to specific requirements.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1572-9699
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The heat activation of trehalase in extracts of sporangiospores of Phycomyces blakesleeanus, following the induction of germination by heat activation and the gelatinization of potato starch granules were studied under different conditions in order to discriminate between several phenomena as possible triggers in the activation of trehalase. Short-chain alcohols (from methanol to pentanol) lower the activation temperature of trehalase while long-chain alcohols (from heptanol to nonanol) raise it. Short-chain alcohols also lower the gelatinization temperature of potato starch granules, while long-chain alcohols, hexanol and heptanol have hardly any influence on the gelatinization temperature. Octanol raises the gelatinization temperature. More polar phenols lower the activation temperature of trehalase, while more apolar phenols will raise it. The gelatinization temperature of starch granules is more lowered by the polar polyphenols than by the more apolar phenols. The effect of high pressure on starch gelatinization was investigated in order to compare data from such a model system with the data on trehalase activation. The gelatinization temperature of starch granules is shifted upwards with about 3–5 K/1000 atm (1.013×105 kPa). Pressures higher than 1500 atm do not further increase the gelatinization temperature. However, no reversal of the effect, as occurs with protein conformational changes, is seen with pressure up to 2500 atm. Also for trehalase activation we find a continuous upward shift of the activation temperature with about 5–9 K/1000 atm. These data are in agreement with a thermal transition in a polysaccharide matrix, being the trigger in the heat activation of trehalase.
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