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© The Rockefeller University Press, 0021-9525/1998//651 $5.00
The Journal of Cell Biology, Volume 142, Number 3, , 1998 651-663

Regular Articles

Retrograde Traffic Out of the Yeast Vacuole to the TGN Occurs via the Prevacuolar/Endosomal Compartment



Nia J. Bryant*, Robert C. Piper{ddagger}, Lois S. Weisman§, and Tom H. Stevens*

* Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229; {ddagger} Department of Physiology, University of Iowa, Iowa City, Iowa 52242; and § Department of Biochemistry, University of Iowa, Iowa City, Iowa 52242

A large number of trafficking steps occur between the last compartment of the Golgi apparatus (TGN) and the vacuole of the yeast Saccharomyces cerevisiae. To date, two intracellular routes from the TGN to the vacuole have been identified. Carboxypeptidase Y (CPY) travels through a prevacuolar/endosomal compartment (PVC), and subsequently on to the vacuole, while alkaline phosphatase (ALP) bypasses this compartment to reach the same organelle. Proteins resident to the TGN achieve their localization despite a continuous flux of traffic by continually being retrieved from the distal PVC by virtue of an aromatic amino acid–containing sorting motif. In this study we report that a hybrid protein based on ALP and containing this retrieval motif reaches the PVC not by following the CPY sorting pathway, but instead by signal-dependent retrograde transport from the vacuole, an organelle previously thought of as a terminal compartment. In addition, we show that a mutation in VAC7, a gene previously identified as being required for vacuolar inheritance, blocks this trafficking step. Finally we show that Vti1p, a v-SNARE required for the delivery of both CPY and ALP to the vacuole, uses retrograde transport out of the vacuole as part of its normal cellular itinerary.

Key Words: endosome • SNARE • TGN • vacuole • VPS

Abbreviations used in this paper: ALP, alkaline phosphatase; CPY, carboxypeptidase Y; DIC, differential interference contrast; DPAP, dipeptidyl aminopeptidase; PVC, prevacuolar/endosomal compartment; SNARE, SNAP receptor; V-ATPase, vacuolar ATPase; VAC, vacuolar inheritance; VPS, vacuolar protein sorting.

Address all correspondence to T.H. Stevens, Institute of Molecular Biology, University of Oregon, Eugene, OR 97403-1229. Tel.: (541) 346-5884. Fax: (541) 346-4854. E-mail: stevens{at}molbio.uoregon.edu


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