Dsl1p, Tip20p, and the novel Dsl3(Sec39) protein are required for the stability of the Q/t-SNARE complex at the endoplasmic reticulum in yeast

The "Dsl1p complex" in Saccharomyces cerevisiae, consisting of Dsl1p and Tip20p, is involved in Golgi-ER retrograde transport and it is functionally conserved from yeast to mammalian cells. To further characterize this complex, we analyzed the function of Dsl3p, a protein that interacts wi...

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Bibliographic Details
Published in:Molecular biology of the cell 2005-09, Vol.16 (9), p.3963-3977
Main Authors: Kraynack, Bryan A, Chan, Angela, Rosenthal, Eva, Essid, Miriam, Umansky, Barbara, Waters, M Gerard, Schmitt, Hans Dieter
Format: Article
Language:English
Subjects:
Carrier Proteins - physiology
Endoplasmic Reticulum - physiology
Glycoproteins - physiology
Membrane Proteins - genetics
Membrane Proteins - physiology
Mutation
Polymerase Chain Reaction
Saccharomyces cerevisiae
Saccharomyces cerevisiae - metabolism
Saccharomyces cerevisiae Proteins - genetics
Saccharomyces cerevisiae Proteins - isolation & purification
Saccharomyces cerevisiae Proteins - physiology
Vesicular Transport Proteins
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Summary:The "Dsl1p complex" in Saccharomyces cerevisiae, consisting of Dsl1p and Tip20p, is involved in Golgi-ER retrograde transport and it is functionally conserved from yeast to mammalian cells. To further characterize this complex, we analyzed the function of Dsl3p, a protein that interacts with Dsl1p in yeast two hybrids screens. DSL3, recently identified in a genome wide analysis of essential genes as SEC39, encodes a cytosolic protein of 82 kDa that is peripherally associated with membranes derived from the ER. There is strong genetic interaction between DSL3 and other factors required for Golgi-ER retrograde transport. Size exclusion chromatography and affinity purification approaches confirmed that Dsl3p is associated with subunits of the "Dsl1p complex." The complex also includes the Q/t-SNARE proteins, Use1p, Sec20p, and Ufe1p, integral membrane proteins that constitute the trimeric acceptor for R/v-SNAREs on Golgi-derived vesicles at the ER. Using mutants, we performed a detailed analysis of interactions between subunits of the Dsl1p complex and the ER-localized SNARE proteins. This analysis showed that both Dsl1p and Dsl3p are required for the stable interaction of the SNARE Use1p with a central subcomplex consisting of Tip20p and the SNARE proteins Ufe1p and Sec20p.
ISSN:1059-1524
1939-4586
1059-1524
DOI:10.1091/mbc.E05-01-0056