Nitrogen regulation of Saccharomyces cerevisiae invertase. Role of the URE2 gene.
The regulation of extracellular enzymes is of great biotechnological interest. We studied the regulatory role of the URE2 gene on the periplasmic invertase of Saccharomyces cerevisiae, because its periplasmic asparaginase is regulated by the URE2/GLN3 system. Enzymatic activity was measured in the isogenic strains P40-1B, the ure2 mutant P40-3C, and the P40-3C strain transformed with the pIC-CS plasmid carrying the URE2 gene. The assays were performed using midlog and stationary phase cells and nitrogen-starved cells from these growth phases. During exponential growth, the level of invertase in both wild-type and ure2 mutant cells was comparable. However, the invertase activity in ure2 mutant cells from stationary phase was sixfold lower than in the wild-type cells. When P40-3C cells were transformed with the pIC-CS plasmid, the wild-type phenotype was restored. On nitrogen starvation in the presence of sucrose, the invertase activity in wild-type cells from midlog phase decreased three times, whereas in stationary cells, the activity decreased eight times. However, invertase activity doubled in ure2 mutant cells from both phases. When these cells were transformed with the aforementioned plasmid, the wild-type phenotype was restored, although a significant invertase decrease in stationary cell was not observed. These results suggested that the URE2 protein plays a role in invertase activity.
Silveira
M C
MC
Instituto de Química, Universidade Federal do Rio de Janeiro, Brazil.
Oliveira
E M
EM
Carvajal
E
E
Bon
E P
EP
D001215:Asparaginase / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D003470:Culture Media; D004926:Escherichia coli; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005979:Glutathione Peroxidase; D006026:Glycoside Hydrolases / Q000235:genetics / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism*; D011328:Prions; D011994:Recombinant Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins; D043324:beta-Fructofuranosidase
D003470:Culture Media; D005656:Fungal Proteins; D011328:Prions; D011994:Recombinant Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D005979:Glutathione Peroxidase; C067020:URE2 protein, S cerevisiae; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D001215:Asparaginase; D009584:Nitrogen