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Published online 18 September 2000. doi:10.1083/jcb.150.6.1263
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© The Rockefeller University Press, 0021-9525/2000//1263 $5.00
The Journal of Cell Biology, Volume 150, Number 6, , 2000 1263-1270

Original Article

Exclusion of Golgi Residents from Transport Vesicles Budding from Golgi Cisternae in Intact Cells



Lelio Orcia, Mylène Amherdta, Mariella Ravazzolaa, Alain Perreleta, and James E. Rothmanb

a Department of Morphology, University of Geneva Medical School, 1211 Geneva 4, Switzerland
b Cellular Biochemistry and Biophysics Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10021

A central feature of cisternal progression/maturation models for anterograde transport across the Golgi stack is the requirement that the entire population of steady-state residents of this organelle be continuously transported backward to earlier cisternae to avoid loss of these residents as the membrane of the oldest (trans-most) cisterna departs the stack. For this to occur, resident proteins must be packaged into retrograde-directed transport vesicles, and to occur at the rate of anterograde transport, resident proteins must be present in vesicles at a higher concentration than in cisternal membranes. We have tested this prediction by localizing two steady-state residents of medial Golgi cisternae (mannosidase II and N-acetylglucosaminyl transferase I) at the electron microscopic level in intact cells. In both cases, these abundant cisternal constituents were strongly excluded from buds and vesicles. This result suggests that cisternal progression takes place substantially more slowly than most protein transport and therefore is unlikely to be the predominant mechanism of anterograde movement.

Key Words: secretion • cisternal maturation • coatomer • cargo • glycosyltransferase

© 2000 The Rockefeller University Press

Abbreviations used in this paper: Man II, mannosidase II; NAGT I, N-acetylglucosaminyltransferase I.


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