The vacuolar protein-targeting signal of yeast carboxypeptidase is functional in oocytes from Xenopus laevis.
Carboxypeptidase Y, a yeast vacuolar glycoprotein was expressed in oocytes from Xenopus laevis and its biosynthesis and sorting were examined. In yeast, targeting to the vacuole, the functional equivalent of the lysosome, is not mannose-6-phosphate-receptor dependent. It was found that carboxypeptidase enters the secretory pathway of the oocyte and is there glycosylated, phosphorylated in the carbohydrate part and delivered to the lysosome. Deletion of an amino acid sequence, previously shown to determine intracellular targeting of this enzyme in yeast, caused a loss of phosphorylation and mislocalization of carboxypeptidase Y into the oocyte medium. Inhibition of glycosylation of carboxypeptidase by tunicamycin did not lead to its secretion. In-frame fusion of the targeting domain to a secretory yeast glycoprotein, invertase, did not prevent its secretion. However, a hybrid containing 80% carboxypeptidase abolished invertase secretion. The results indicate that the vacuolar protein-targeting signal from yeast carboxypeptidase can, in principal, function in a higher eukaryote.
Roitsch
T
T
Fakultät für Biologie und Vorklinische Medizin, Universität Regensburg, Federal Republic of Germany.
Lehle
L
L
D000118:Acetylglucosaminidase / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D002268:Carboxypeptidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D043402:Cathepsin A; D002872:Chromosome Deletion; D005260:Female; D008247:Lysosomes / Q000201:enzymology; D009865:Oocytes / Q000201:enzymology*; D010957:Plasmids; D014176:Protein Biosynthesis; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism*; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D014158:Transcription, Genetic; D014617:Vacuoles / Q000201:enzymology*; D014982:Xenopus laevis
D021382:Protein Sorting Signals; D029701:Saccharomyces cerevisiae Proteins; D000118:Acetylglucosaminidase; D002268:Carboxypeptidases; D043402:Cathepsin A; C499283:PRC1 protein, S cerevisiae; C022633:serine carboxypeptidase