Formation of AP-3 transport intermediates requires Vps41 function

Peter Rehling; Tamara Darsow; David J. Katzmann; Scott D. Emr

doi:10.1038/14037

Formation of AP-3 transport intermediates requires Vps41 function

Peter Rehling, Tamara Darsow, David J. Katzmann, Scott D. Emr

Biochemistry and Molecular Biology

Research output: Contribution to journal › Article › peer-review

96 Scopus citations

Abstract

Transport of a subset of membrane proteins to the yeast vacuole requires the function of the AP-3 adaptor protein complex. To define the molecular requirements of vesicular transport in this pathway, we used a biochemical approach to analyse the formation and content of the AP-3 transport intermediate. A vam3^tsf (vacuolar t-SNARE) mutant blocks vesicle docking and fusion with the vacuole and causes the accumulation of 50-130-nanometre membrane vesicles, which we isolated and showed by biochemical analysis and immunocytochemistry to contain both AP-3 adaptors and alkaline phosphatase (ALP) pathway cargoes. Inactivation of AP-3 or the protein Vps41 blocks formation of this vesicular intermediate. Vps41 binds to the AP-3 δ-adaptin subunit, suggesting that they function together in the formation of ALP pathway transport intermediates at the late Golgi.

Original language	English (US)
Pages (from-to)	346-353
Number of pages	8
Journal	Nature Cell Biology
Volume	1
Issue number	6
DOIs	https://doi.org/10.1038/14037
State	Published - Oct 1999

ASJC Scopus subject areas

Cell Biology

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title = "Formation of AP-3 transport intermediates requires Vps41 function",

abstract = "Transport of a subset of membrane proteins to the yeast vacuole requires the function of the AP-3 adaptor protein complex. To define the molecular requirements of vesicular transport in this pathway, we used a biochemical approach to analyse the formation and content of the AP-3 transport intermediate. A vam3tsf (vacuolar t-SNARE) mutant blocks vesicle docking and fusion with the vacuole and causes the accumulation of 50-130-nanometre membrane vesicles, which we isolated and showed by biochemical analysis and immunocytochemistry to contain both AP-3 adaptors and alkaline phosphatase (ALP) pathway cargoes. Inactivation of AP-3 or the protein Vps41 blocks formation of this vesicular intermediate. Vps41 binds to the AP-3 δ-adaptin subunit, suggesting that they function together in the formation of ALP pathway transport intermediates at the late Golgi.",

author = "Peter Rehling and Tamara Darsow and Katzmann, {David J.} and Emr, {Scott D.}",

note = "Funding Information: ACKNOWLEDGEMENTS We thank T. McQuistan and J.M. McCaffery for expert assistance with electron microscopy analysis. Affinity-purified Kex2 antibodies were a gift from G. Payne. We thank members of the Emr laboratory for helpful discussions, particularly A. Wurmser, J. Gary and M. Babst for critical reading of the manuscript. P.R. is supported by a postdoctoral fellowship from the Deutsche Forschungsgemeinschaft (Re 1384/1-1) and D.J.K. is an Associate of the Howard Hughes Medical Institute. This work was supported by NIH grant CA58689 (to S.D.E.). S.D.E. is an Investigator with the Howard Hughes Medical Institute. Correspondence and requests for materials should be addressed to S.D.E.",

year = "1999",

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doi = "10.1038/14037",

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AU - Katzmann, David J.

AU - Emr, Scott D.

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PY - 1999/10

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N2 - Transport of a subset of membrane proteins to the yeast vacuole requires the function of the AP-3 adaptor protein complex. To define the molecular requirements of vesicular transport in this pathway, we used a biochemical approach to analyse the formation and content of the AP-3 transport intermediate. A vam3tsf (vacuolar t-SNARE) mutant blocks vesicle docking and fusion with the vacuole and causes the accumulation of 50-130-nanometre membrane vesicles, which we isolated and showed by biochemical analysis and immunocytochemistry to contain both AP-3 adaptors and alkaline phosphatase (ALP) pathway cargoes. Inactivation of AP-3 or the protein Vps41 blocks formation of this vesicular intermediate. Vps41 binds to the AP-3 δ-adaptin subunit, suggesting that they function together in the formation of ALP pathway transport intermediates at the late Golgi.

AB - Transport of a subset of membrane proteins to the yeast vacuole requires the function of the AP-3 adaptor protein complex. To define the molecular requirements of vesicular transport in this pathway, we used a biochemical approach to analyse the formation and content of the AP-3 transport intermediate. A vam3tsf (vacuolar t-SNARE) mutant blocks vesicle docking and fusion with the vacuole and causes the accumulation of 50-130-nanometre membrane vesicles, which we isolated and showed by biochemical analysis and immunocytochemistry to contain both AP-3 adaptors and alkaline phosphatase (ALP) pathway cargoes. Inactivation of AP-3 or the protein Vps41 blocks formation of this vesicular intermediate. Vps41 binds to the AP-3 δ-adaptin subunit, suggesting that they function together in the formation of ALP pathway transport intermediates at the late Golgi.

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Formation of AP-3 transport intermediates requires Vps41 function

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