Analysis of glucose repression in Saccharomyces cerevisiae by pulsing glucose to a galactose-limited continuous culture.
In this study, glucose repression in Saccharomyces cerevisiae was analysed under defined physiological conditions, at both the molecular and physiological levels, by pulsing glucose to a galactose-limited continuous culture. During this pulse of glucose, the galactose feed was kept constant. Directly after the glucose pulse, carbon dioxide production increased while oxygen consumption remained constant, demonstrating that the surplus of glucose had been consumed by means of fermentation. The direct accumulation of galactose in the medium after the glucose pulse indicated that the consumption of galactose had been stopped instantaneously. Galactose uptake experiments revealed that the galactose transporter was still present but apparently was incapable of galactose uptake, which could be due to inhibition of the galactose transporter by glucose. The total concentration of cAMP increased from 5 nmol g-1 at t = 0 to 25 nmol g-1 at t = 1.5 min. After 2 min the concentration of cAMP gradually decreased again to the normal level. Within 2 min after the addition of glucose, the transcription of the GAL genes and SUC2 was inhibited. In addition, the transcription of the HXK1 gene, encoding hexokinase isoenzyme 1, was also inhibited, which demonstrates that the HXK1 gene is regulated at the transcriptional level comparable with invertase.
Sierkstra
L N
LN
Department of Molecular Cell Biology, University of Utrecht, The Netherlands.
Nouwen
N P
NP
Verbakel
J M
JM
Verrips
C T
CT
D000255:Adenosine Triphosphate / Q000032:analysis; D001483:Base Sequence; D001692:Biological Transport; D016384:Consensus Sequence; D000242:Cyclic AMP / Q000032:analysis; D004794:Enzyme Repression; D005636:Fructosephosphates / Q000032:analysis; D005690:Galactose / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism* / Q000494:pharmacology; D019298:Glucose-6-Phosphate; D005958:Glucosephosphates / Q000032:analysis; D006593:Hexokinase / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development
D005636:Fructosephosphates; D005958:Glucosephosphates; D012333:RNA, Messenger; D019298:Glucose-6-Phosphate; C027618:fructose-6-phosphate; D000255:Adenosine Triphosphate; D000242:Cyclic AMP; D006593:Hexokinase; D005947:Glucose; D005690:Galactose