Protein synthesis during transition and stationary phases under glucose limitation in Saccharomyces cerevisiae

Boucherie, Hélian

doi:10.1128/jb.161.1.385-392.1985

Cited by 102 publications

(41 citation statements)

References 32 publications

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“…HSP70 proteins are found in cells under all conditions, presumably due to the central role they play in processes involving folding, assembly and transport of proteins across membranes, as well as the recognition and disaggregation of misfolded or foreign proteins (reviewed by Bienz andby Lindquist andCraig, 1988). A subset of heat shock proteins is expressed as S. cerevisiae progresses into stationary phase (Boucherie, 1985) and enter the Go arrest stage (lida and Yahara, 1984). SSA3 is the only member of the large yeast HSP70 multigene family known to become more actively expressed after the diauxic shift.…”

Section: Discussionmentioning

confidence: 99%

“…When the cells can no longer continue to divide, they become arrested in an unbudded state at START A, analogous to the Go state of the mammalian cell cycle. Cells arrested in Go are metabolically active and express a small, characteristic set of proteins including a subset of heat shock proteins (Iida and Yahara, 1984;Boucherie, 1985). Go arrested cells are constitutively thermotolerant (Schenberg-Frascino and Moustacchi, 1972;Plesset et al, 1987), a phenotype believed to be related to the expression of these heat shock proteins.…”

Section: Oxford University Pressmentioning

confidence: 99%

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Regulation of a yeast HSP70 gene by a cAMP responsive transcriptional control element.

1990

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HSP70 genes exhibit complex regulation in response to stress and a variety of cellular and developmental events. The SSA3 HSP70 gene of Saccharomyces cerevisiae is activated at the transcriptional level under conditions of nutrient limitation. Analysis of deletions revealed that cis‐acting DNA sequences present immediately upstream and downstream of the previously identified heat shock elements (UASHS) mediate this regulation. A 35 bp region of SSA3, distinct from UASHS, contains sequences capable of activating a heterologous promoter following the diauxic shift and in the stationary phase of the yeast life cycle; this region has been designated an upstream activating sequence, UASPDS. Expression driven by UASPDS is regulated by the RAS/cAMP pathway. Reduced cAMP dependent protein kinase activity results in UASPDS dependent activation of the SSA3 promoter while constitutive cAMP dependent protein kinase activity prevents UASPDS mediated transcription, even under growth conditions that would normally result in full activation. Although the heat shock element alone exhibits no UAS activity under conditions in which UASPDS promotes transcription, UASHS interacts positively with UASPDS to mediate high levels of SSA3 transcription in response to nutrient limitation and lowered intracellular cAMP concentration. This interaction is independent of the precise spacing and relative orientation of the two elements.

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Section: Discussionmentioning

confidence: 99%

Section: Oxford University Pressmentioning

confidence: 99%

Regulation of a yeast HSP70 gene by a cAMP responsive transcriptional control element.

1990

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“…In a previous report, we showed that glucose limitation is a particularly suitable condition for approaching questions concerning changes in protein synthesis when yeast cells undergo starvation (Boucherie, 1985). Drastic alterations in the two-dimensional pattern of protein synthesis were observed.…”

Section: Mmsurement Of Cellular Parametersmentioning

confidence: 95%

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confidence: 99%

Induction of a heat‐shock‐type response in Saccharomyces cerevisiae following glucose limitation

Bataillé

Régnacq

Boucherie

1991

Yeast

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The protein pattern of yeast cells which have arrested proliferation in response to glucose exhaustion is drastically different from that of exponentially growing cells (Boucherie, 1985). In this study, we used two-dimensional gel electrophoresis to characterize the protein events responsible for these alterations. We found that the induction of heat-shock proteins is one of the major events responsible for these changes. This induction accounts for the synthesis of 18 of the 35 novel polypeptides observed in glucose-limited cells. It was shown to occur in combination with two other protein events: the derepression of carbon catabolite repressed proteins, which accounts for the synthesis of the other novel polypeptides, and an arrest of the synthesis of almost all the proteins present in exponentially growing cells. The time course of each of these events was determined by carrying out a detailed analysis of the pattern of proteins synthesized at various stages of a culture exhausting its glucose supply, and by the measurement of the rate of synthesis of individual polypeptides. The results showed in particular that the synthesis of most of the heat-shock proteins synthesized in glucose-limited cells was induced closely before glucose exhaustion, and that this synthesis was transient, climaxing by the time glucose was exhausted. Under the culture condition investigated, the entry into stationary phase associated with glucose limitation began several hours before glucose exhaustion. It was thus concluded that the observed induction of heat-shock proteins is directly related to the nutritional limitation and is independent from the arrest of cell proliferation.

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“…Growth of Saccharomyces cerevisiae on glucose elicits a well-known pattern of enzymes and metabolites [l, 21. In the last years some observations have been reported that suggest that this pattern may not be constant during growth. For example a decrease in RNA and protein synthesis is observed well in advance of glucose exhaustion [3]. Also glycogen accumulation begins when about half of the original glucose remains in the culture irrespective of the initial concentration of the sugar [4].…”

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Changes in the concentration of cAMP, fructose 2,6‐bisphosphate and related metabolites and enzymes in Saccharomyces cerevisiae during growth on glucose

François

Eraso

Gancedo

1987

European Journal of Biochemistry

117

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Changes in the concentration of several metabolites and enzymes related to carbohydrate metabolism were measured during the growth of Saccharomyces cerevisiae on a mineral medium containing glucose as the limiting nutrient. When about 50% of the original glucose was used the exponential phase ended and the culture entered a 'transition' phase before the complete exhaustion of glucose. In this transition phase several metabolic changes occurred. CAMP, that decreased along growth, reached a constant value of about 0.7 nmol/g dry weight. A pronounced drop in fructose-6-phosphate-2-kinase activity and in the concentration of fructose 2,6-bisphosphate and fructose 1,6-bisphosphate was observed accompanied by a less marked decrease in hexose monophosphates. Trehalase activity also dropped and reached a minimal value at the onset of the stationary phase when synthesis of trehalose began. Glycogen concentration and glycogen synthase activity increased sharply during the transition phase. Plasma membrane ATPase began to increase at the middle of the exponential phase and then, coincident with the glucose exhaustion, a 90% decrease in the measurable activity was observed.Growth of Saccharomyces cerevisiae on glucose elicits a well-known pattern of enzymes and metabolites [l, 21. In the last years some observations have been reported that suggest that this pattern may not be constant during growth. For example a decrease in RNA and protein synthesis is observed well in advance of glucose exhaustion [3]. Also glycogen accumulation begins when about half of the original glucose remains in the culture irrespective of the initial concentration of the sugar [4].Recently cAMP and fructose 2,6-bisphosphate [Fru(2,6)-P2] have received considerable attention as possible 'organizers' of carbohydrate metabolism in yeast [5, 61. In order to provide more experimental data to disprove or support this idea we initiated a systematic investigation of the concentrations of these compounds and some other metabolites and enzymes related to carbohydrate metabolism during growth of yeast on glucose. The results of our study show that exponential growth ceases when glucose is still present in the culture and that important metabolic changes occur in the transition phase between the exponential and stationary phase of growth. MATERIALS AND METHODS Growth conditionsSaccharomyces cerevisiae X2180 was grown at 30°C in a mineral medium [7] using NaCl instead of the original sodium citrate. The pH of the medium was adjusted to 5.5 with KOH. Glucose was added after sterilization at a final concentration of 2%. The media were inoculated with yeasts grown to the stationary phase on a medium containing 2% glucose, 1% yeast extract and 1 YO bactopeptone. The initial cell concentration was about lo5 cells/ml. Growth was followed by measuring the turbidity of the culture at 660 nm and the corresponding dry weight was calculated from a calibration curve of dry weight versus absorbance (Fig. 1). This curve was constructed in the same conditions as the subseque...

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Protein synthesis during transition and stationary phases under glucose limitation in Saccharomyces cerevisiae

Cited by 102 publications

References 32 publications

Regulation of a yeast HSP70 gene by a cAMP responsive transcriptional control element.

Regulation of a yeast HSP70 gene by a cAMP responsive transcriptional control element.

Induction of a heat‐shock‐type response in Saccharomyces cerevisiae following glucose limitation

Changes in the concentration of cAMP, fructose 2,6‐bisphosphate and related metabolites and enzymes in Saccharomyces cerevisiae during growth on glucose

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