The localization of proteins to late-Golgi membranes (TGN) of Saccharomyces cerevisiae is conferred by targeting motifs containing aromatic residues in the cytosolic domains of these proteins. These signals could act by directing retrieval from a post-Golgi compartment or by preventing exit from the TGN. To investigate the mechanism of localization of yeast TGN proteins, we used the heterologous protein A-ALP (consisting of the cytosolic domain of dipeptidyl aminopeptidase A [DPAP A] fused to the transmembrane and luminal domains of the vacuolar protein alkaline phosphatase [ALP]), which localizes to the yeast TGN. Insertion of the aromatic residue–based TGN localization motif (FXFXD) of DPAP A into the cytosolic domain of ALP results in a protein that resides in the TGN. We demonstrate that the FXFXD motif confers Golgi localization through retrieval from a post-Golgi compartment by detecting a post-Golgi processed form of this protein in the TGN. We present an assay that uncouples retrieval-mediated Golgi localization from static retention-based localization, allowing measurement of the rate at which proteins exit the yeast TGN. We also demonstrate that the cytosolic domain of DPAP A contains additional information, separate from the retrieval motif, that slows exit from the TGN. We propose a model for DPAP A localization that involves two distinct mechanisms: one in which the FXFXD motif directs retrieval from a post-Golgi compartment, and a second that slows the rate at which DPAP A exits the TGN.
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27 January 1997
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January 27 1997
Two Separate Signals Act Independently to Localize a Yeast Late Golgi Membrane Protein through a Combination of Retrieval and Retention
Nia J. Bryant,
Nia J. Bryant
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229
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Tom H. Stevens
Tom H. Stevens
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229
Search for other works by this author on:
Nia J. Bryant
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229
Tom H. Stevens
Institute of Molecular Biology, University of Oregon, Eugene, Oregon 97403-1229
Address all correspondence to Tom H. Stevens, Institute of Molecular Biology, University of Oregon, Eugene, OR 97403-1229. Tel.: (541) 3465884. Fax: (541) 346-4854. e-mail: [email protected]
Received:
September 09 1996
Revision Received:
November 01 1996
Online ISSN: 1540-8140
Print ISSN: 0021-9525
1997
J Cell Biol (1997) 136 (2): 287–297.
Article history
Received:
September 09 1996
Revision Received:
November 01 1996
Citation
Nia J. Bryant, Tom H. Stevens; Two Separate Signals Act Independently to Localize a Yeast Late Golgi Membrane Protein through a Combination of Retrieval and Retention. J Cell Biol 27 January 1997; 136 (2): 287–297. doi: https://doi.org/10.1083/jcb.136.2.287
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