Vesicular transport of extracellular acid phosphatases in yeast Saccharomyces cerevisiae.
A method for isolation of secretory vesicles from the yeast Saccharomyces cerevisiae based on the disintegration of protoplasts by osmotic shock followed by separation of the vesicles by centrifugation in a density gradient of Urografin was developed in this study. Two populations of the secretory vesicles that differ in density and shape were separated. Acid phosphatases (EC 3.1.3.2) were used as markers of the secretory vesicles. It was shown that the constitutive acid phosphatase (PHO3 gene product) is mainly transported to the cell surface by a lower density population of vesicles, while the repressible acid phosphatase (a heteromer encoded by PHO5, PHO10, and PHO11 genes) by a vesicle population of higher density. These data provide evidence that at least two pathways of transport of yeast secretory proteins from the place of their synthesis and maturation to the cell surface may exist. To reveal the probable reasons for transport of Pho3p and Pho5p/Pho10p/Pho11p enzymes by two different kinds of vesicles, we isolated vesicles from strains that synthesize the homomeric forms of the repressible acid phosphatase. It was demonstrated that glycoproteins encoded by the PHO10 and/or PHO11 genes could be responsible for the choice of one of the alternative transport pathways of the repressible acid phosphatase. A high correlation coefficient between bud formation and secretion of Pho5p phosphatase and the absence of correlation between bud formation and secretion of minor phosphatases Pho10p and Pho11p suggests different functional roles of the polypeptides that constitute the native repressible acid phosphatase.
Blinnikova
E I
EI
Department of Molecular Biology, School of Biology, Lomonosov Moscow State University, Moscow, 119899, Russia.
Mirjuschenko
F L
FL
Shabalin
Yu A
YA
Egorov
S N
SN
D000135:Acid Phosphatase / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport; D005110:Extracellular Space / Q000378:metabolism; D005800:Genes, Fungal; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D022142:Secretory Vesicles / Q000378:metabolism*
D000135:Acid Phosphatase