title: Ultracytochemical evidence of Golgi functions in microvesicles at all phases of cell cycle in Saccharomyces cerevisiae.
abstract: The topical question of Golgi compartment identity in the ascomycetous yeast Saccharomyces cerevisiae is illustrated by a multiple ultracytochemical approach. For this eucaryotic single-cell organism the established scheme of secretory transport via a cascade of cisternae housing different functions of Golgi apparatus has been deduced principally of genetic and molecular analyses ex situ and confirms the mammalian secretion scheme. Nevertheless, ultracytochemical in situ localizations of enzyme activities engaged in secretion represented evidence for localization of important steps of secretory glycoprotein maturation in two morphologically distinct populations of transport microvesicles formed from endoplasmic reticulum and Golgi cisternae. Both types of microvesicles function in exocytosis or transport into lysosomal vacuoles and have identical charge. However, their presence differs in interphase and in budding cells of S. cerevisiae. Smooth, larger membrane bound microvesicles are conspicuous at the onset of budding and at construction of scars, while the coated, smaller microvesicles of globular ultrastructure are present constitutively, throughout the cell cycle. Because the established model of the yeast secretory path considers only the part of the budding phase preceding the onset of mitosis, an alternative scheme for the cellular mechanism of glycoprotein secretion in S. cerevisiae that distinguishes interphase and budding yeast, has been established. The lumen of microvesicles contains proteases catalysing maturation of the mating pheromone alpha-factor (yscIV, yscF), vacuolar protease yscY, alkaline phosphohydrolase, polyphosphorylated components of the bud scar and glycoproteins. The in situ approach also reveals a minimum level of alpha-factor precursor processing proteolytic activity at the budding phase of cells, a transient presence of polyphosphorylated compounds in the bud scars and their transport by microvesicles. Ultracytochemical reactions suggest that the nuclear envelope lumen houses certain functions attributed to endoplasmic reticulum and that some steps of outer-chain glycosylation may occur in microvesicles. Microvesicles which contain proteases and polyphosphorylated intermediates also appear in juvenile vacuoles (lysosomes). Ultracytochemical findings show the Golgi compartment of S. cerevisiae to consist not only of discrete endoplasmic cisternae, immunodetected by others as sites of outer chain alpha-1,6-mannosylation and of the Golgi membrane marker proteins Sec7p and Ypt1p, but also of microvesicles moving either to the cell plasma membrane or to vacuoles. The previously hypothesized hierarchy of segregated yeast Golgi cisternae was not revealed by ultracytochemical findings.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Institute of Microbiology, Academy of Sciences of Czech Republic, Praha.
authors.forename: J
authors.initials: J
authors.lastname: Vorísek
mesh_terms: D000626:Aminopeptidases / Q000378:metabolism; D000818:Animals; D002453:Cell Cycle / Q000502:physiology; D005089:Exocytosis / Q000502:physiology; D006023:Glycoproteins / Q000378:metabolism; D006056:Golgi Apparatus / Q000502:physiology* / Q000648:ultrastructure; D006651:Histocytochemistry; D016253:Microscopy, Immunoelectron; D015388:Organelles / Q000502:physiology* / Q000648:ultrastructure; D043484:Proprotein Convertases; D012441:Saccharomyces cerevisiae / Q000502:physiology* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins; D013381:Subtilisins / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D029701:Saccharomyces cerevisiae Proteins; D000626:Aminopeptidases; C022750:X-Pro aminopeptidase; D043484:Proprotein Convertases; D013381:Subtilisins; C058751:KEX2 protein, S cerevisiae