Control of glucose influx into glycolysis and pleiotropic effects studied in different isogenic sets of Saccharomyces cerevisiae mutants in trehalose biosynthesis.
The GGS1/TPS1 gene of the yeast Saccharomyces cerevisiae encodes the trehalose-6-phosphate synthase subunit of the trehalose synthase complex. Mutants defective in GGS1/TPS1 have been isolated repeatedly and they showed variable pleiotropic phenotypes, in particular with respect to trehalose content, ability to grow on fermentable sugars, glucose-induced signaling and sporulation capacity. We have introduced the fdp1, cif1, byp1 and glc6 alleles and the ggs1/tps1 deletion into three different wild-type strains, M5, SP1 and W303-1A. This set of strains will aid further studies on the molecular basis of the complex pleiotropic phenotypes of ggs1/tps1 mutants. The phenotypes conferred by specific alleles were clearly dependent on the genetic background and also differed for some of the alleles. Our results show that the lethality caused by single gene deletion in one genetic background can become undetectable in another background. The sporulation defect of ggs1/tps1 diploids was neither due to a deficiency in G1 arrest, nor to the inability to accumulate trehalose. Ggs1/tps1 delta mutants were very sensitive to glucose and fructose, even in the presence of a 100-fold higher galactose concentration. Fifty-percent inhibition occurred at concentrations similar to the Km values of glucose and fructose transport. The inhibitory effect of glucose in the presence of a large excess of galactose argues against an overactive glycolytic flux as the cause of the growth defect. Deletion of genes of the glucose carrier family shifted the 50% growth inhibition to higher sugar concentrations. This finding allows for a novel approach to estimate the relevance of the many putative glucose carrier genes in S. cerevisiae. We also show that the GGS1/TPS1 gene product is not only required for the transition from respirative to fermentative metabolism but continuously during logarithmic growth on glucose, in spite of the absence of trehalose under such conditions.
Neves
M J
MJ
Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit te Leuven, Leuven-Heverlee, Belgium.
Hohmann
S
S
Bell
W
W
Dumortier
F
F
Luyten
K
K
Ramos
J
J
Cobbaert
P
P
de Koning
W
W
Kaneva
Z
Z
Thevelein
J M
JM
D002455:Cell Division; D004247:DNA / Q000096:biosynthesis; D005632:Fructose / Q000378:metabolism; D006597:Fructose-Bisphosphatase / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D019298:Glucose-6-Phosphate; D005958:Glucosephosphates / Q000378:metabolism; D005964:Glucosyltransferases / Q000235:genetics / Q000378:metabolism*; D006006:Glycogen Synthase / Q000378:metabolism; D006019:Glycolysis; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D013172:Spores, Fungal; D014199:Trehalose / Q000096:biosynthesis*
D005958:Glucosephosphates; D005632:Fructose; D019298:Glucose-6-Phosphate; D004247:DNA; D014199:Trehalose; D005964:Glucosyltransferases; D006006:Glycogen Synthase; C023180:trehalose-6-phosphate synthase; D006597:Fructose-Bisphosphatase; D005947:Glucose