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Journal of Cell Science 114, 2231-2239 (2001)
© 2001 The Company of Biologists Limited


RESEARCH ARTICLE

Three dimensional configuration of the secretory pathway and segregation of secretion granules in the yeast Saccharomyces cerevisiae

Alain Rambourg1, Catherine L. Jackson2,* and Yves Clermont3

1 Département de Biologie Cellulaire et Moléculaire, CEA/Saclay, F-91191 Gif-sur-Yvette, France
2 Cell Biology and Metabolism Branch, NICHD, NIH, Bldg. 18T, Room 101, 18 Library Drive, Bethesda, MD 20892-5430, USA
3 Department of Anatomy and Cell Biology, McGill University, 3640 University Street, Montréal, Québec, H3A 2B2, Canada

*Author for correspondence (e-mail: cathyj{at}helix.nih.gov)

Accepted March 3, 2001

The structural elements of the secretory pathway in the budding yeast Saccharomyces cerevisiae were analyzed by 3D stereo-electron microscopy using relatively thick sections in which membranes were selectively impregnated. In a wild-type strain, tubular networks of various sizes and staining properties were distributed throughout the cytoplasm. As a rule, wide-meshed, lightly stained polygonal networks were connected to more or less fenestrated sheets of endoplasmic reticulum (ER). Some of these networks were continuous with more intensely stained networks and narrower meshes that displayed at their intersections nodular dilations that progressively increased in size and staining properties to reach those of secretion granules. Such networks presumably corresponded to Golgi elements. Indeed, stacked cisternae typical of the mammalian Golgi apparatus are rarely found in wild-type cells. However, if it is assumed that the Golgi apparatus plays a key role in the segregation and maturation of secretion granules, then tubular networks with nodular dilations should be equivalent to parts of this organelle. In correlation with the increase in size and density of the nodules there was a decrease in diameter and staining intensity of the interconnecting tubules. These results parallel observations on the formation of secretory granules in mammalian cells and suggest that the segregation of secretory material is concomitant with the progressive perforation and tubulization of previously unperforated sheets. When the sec21-3 thermosensitive mutant was examined at the nonpermissive temperature (37°C), the secretory pathway was blocked at exit from the ER, which started to accumulate as clusters of narrow, anastomosed, unperforated ribbon-like elements. When the block was released by shifting down to permissive temperature (24°C), tubular networks of various sizes and caliber, presumably Golgi in nature, formed as soon as 5 minutes after release of the block. At later time intervals, granules of various sizes and densities appeared to be released by rupture of these tubular networks or even to form at the edges of ER fenestrae. These observations support a dynamic maturation process in which the formation of secretion granules occurs by means of an oriented series of membrane transformations starting at the ER and culminating with the liberation of secretion granules from Golgi networks.

Key words: Secretory pathway, Golgi apparatus, Secretion granules, Yeast, Brefeldin A, BFA, Trafficking


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A. Peyroche, R. Courbeyrette, A. Rambourg, and C. L. Jackson
The ARF exchange factors Gea1p and Gea2p regulate Golgi structure and function in yeast
J. Cell Sci., March 8, 2002; 114(12): 2241 - 2253.
[Abstract] [Full Text] [PDF]




© The Company of Biologists Ltd 2001