mBet3p is required for homotypic COPII vesicle tethering in mammalian cells (original) (raw)
Article| July 31 2006
1Howard Hughes Medical Institute
2Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06519
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2Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06519
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2Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06519
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3Division of Biological Sciences, The University of Montana, Missoula, MT 59812
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3Division of Biological Sciences, The University of Montana, Missoula, MT 59812
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1Howard Hughes Medical Institute
2Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06519
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Sidney Yu
1Howard Hughes Medical Institute
2Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06519
Ayano Satoh
2Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06519
Marc Pypaert
2Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06519
Karl Mullen
3Division of Biological Sciences, The University of Montana, Missoula, MT 59812
Jesse C. Hay
3Division of Biological Sciences, The University of Montana, Missoula, MT 59812
Susan Ferro-Novick
1Howard Hughes Medical Institute
2Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06519
Abbreviations used in this paper: MVB, multivesicular body; tER, transitional ER; VTC, vesicular tubular cluster.
Received: March 09 2006
Accepted: June 23 2006
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2006
J Cell Biol (2006) 174 (3): 359–368.
TRAPPI is a large complex that mediates the tethering of COPII vesicles to the Golgi (heterotypic tethering) in the yeast Saccharomyces cerevisiae. In mammalian cells, COPII vesicles derived from the transitional endoplasmic reticulum (tER) do not tether directly to the Golgi, instead, they appear to tether to each other (homotypic tethering) to form vesicular tubular clusters (VTCs). We show that mammalian Bet3p (mBet3p), which is the most highly conserved TRAPP subunit, resides on the tER and adjacent VTCs. The inactivation of mBet3p results in the accumulation of cargo in membranes that colocalize with the COPII coat. Furthermore, using an assay that reconstitutes VTC biogenesis in vitro, we demonstrate that mBet3p is required for the tethering and fusion of COPII vesicles to each other. Consistent with the proposal that mBet3p is required for VTC biogenesis, we find that ERGIC-53 (VTC marker) and Golgi architecture are disrupted in siRNA-treated mBet3p-depleted cells. These findings imply that the TRAPPI complex is essential for VTC biogenesis.
The Rockefeller University Press
2006
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